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Biotek MultiFlo FX, Operator'S Manual

The Biotek MultiFlo FX Operator's Manual is a comprehensive guide for using our state-of-the-art laboratory instrument. This essential manual can be easily downloaded for free from manualshive.com, providing users with step-by-step instructions and detailed information on optimizing their research workflow.

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Biomagnetic Separation - Magnetic Bead Assays | 145

Biomagnetic Separation - Magnetic Bead Assays

The microplate carrier, when the magnet adapter is used, supports placement of a
magnet under the microplate. The magnet induces magnetic beads to settle at the
bottom of the wells to help retain them during a wash protocol’s aspirate cycle.

The MultiFlo FX supports standard microplates and these magnets available for
purchase from BioTek:

Plate Type Magnet

PN

96-well

96 Flat Magnet

7103016

96 Ring Magnet

7102216

384-well

384 Flat Magnet

7103017

384 Ring Magnet 7102215

You can use a different magnet if it fits in the carrier and accommodates your
plates. Contact BioTek TAC or visit the Customer Resource Center at
www.biotek.com to obtain a drawing of the carrier with its dimensions.

Persons with pacemakers/implants should avoid direct contact.

Keep all

magnetic media, watches, and sensitive electronic devices away from the magnets.
Credit cards, tapes, and disks can be erased in the presence of a magnetic field. Bodily
harm [pinching of hands and skin] can result if magnets are not handled correctly.
Maintain distance between two or more magnets.

Take advantage of the sample magnetic-bead protocols shipped onboard the MultiFlo

FX. Copy W-Luminex_Mag_Flat_96, for example, and modify the parameters to suit your
assay requirements.

Handling and Cleaning the Magnets

For best magnet strength and bead retention, the bottom of the microplate must be
as close to the magnet as possible. We recommend using flat-bottom plates with
minimal support “webbing” between the sides of the outer wells and the plate
skirts.

Handle the magnets with care. Avoid direct contact with the magnet material. Keep
loose ferrous material away and do not attempt to disassemble.

BioTek Instruments, Inc.

Summary of Contents for MultiFlo FX

Page 1: ...Operator s Manual Microplate Dispenser MultiFlo FX Microplate Dispenser ...

Page 2: ......

Page 3: ...MultiFlo FX Multi Mode Dispenser Operator s Manual BioTek Instruments Inc May 2014 May 2014 Part Number 1261000 Revision B ...

Page 4: ...demark of BioTek Instruments Inc Microsoft Internet Explorer Windows Windows 7 Windows 8 Windows Vista Windows XP and Excel are either registered trademarks or trademarks of Microsoft Corporation in the United States and or other countries All other trademarks are the property of their respective holders Restrictions and Liabilities Information in this document is subject to change and does not re...

Page 5: ...aste Electrical and Electronic Equipment xvii Directive 98 79 EC In Vitro Diagnostics if labeled for this use xvii Electromagnetic Interference and Susceptibility xvii USA FCC CLASS A xvii Canadian Department of Communications Class A xviii User Safety xviii Safety Symbols xix Introduction 1 Introducing the MultiFlo FX Dispenser 2 Features of the MultiFlo FX 3 Washer 3 Peri pump Dispenser 4 Syring...

Page 6: ...Peri Pump 46 Install RAD Technology Components 51 Install the Tubing Bracket and RAD Module 51 Install the tubing bracket 52 Attach the RAD Module to the Tubing Bracket 52 Install RAD Dispense Head 53 Dispense Cassette Diagram 54 Install the Dispense Cassette 56 Install RAD Technology Cassette 58 Prime Trough Inserts 59 Peri pump Reservoir Holder 60 Shorten the Dispense Cassette Tubing 61 Install ...

Page 7: ...ocols from a memory stick USB drive 96 Hardware Change Detected 96 Optimize Performance 98 Recommended prime volumes for the Strip Washer 99 Recommended prime volumes for the Peri pump 99 Recommended prime volumes for the Syringe dispensers 101 Optimize protocols to improve evacuation 101 Add a secondary aspiration to a wash cycle 101 Create or Modify a Protocol 102 Protocol Parameters Tables 103 ...

Page 8: ...aration Magnetic Bead Assays 145 Perform Magnetic Bead Assays 146 Optimize Magnetic Bead Protocols 147 Flat Magnets 147 Ring Magnets 148 How to determine the Magnet Adapter Height Offset 149 Magnet Height Offset 150 Special Procedure for Magnetic Bead Assays 151 Peri pump Peristaltic Dispenser 152 Peri pump How it works 153 Release the tension on the dispense cassette 154 Peri pump Dispense Step 1...

Page 9: ...he Plate Carrier 191 Clean the exterior surfaces 191 Peri pump Dispenser Maintenance 192 Flush the Dispense Cassette 192 Unclog the Dispense Tips 194 RAD Single tube Cassette Unclog the Tip 196 RAD 8 to 1 Cassette Unclog the Tips 197 Record the Number of Plates Processed 199 RAD 8 to 1 Cassette Replace Chute 200 Syringe Dispenser and Strip Washer Maintenance 201 Clean the Manifolds 201 Clean the D...

Page 10: ... Precision Accuracy Test 236 Washer Qualification Liquid Tests 250 Dispense Precision and Accuracy Test 250 Evacuation Efficiency Test 250 Washer Qualification Test Materials 251 Create Washer Qualification Protocols 252 Dispense Precision and Accuracy Test 253 Evacuation Efficiency Test 254 Evacuation Diagnostic Test 256 Troubleshooting 271 Troubleshooting 272 Syringe Dispenser Troubleshooting 27...

Page 11: ...4 P R China Phone 800 242 4685 Fax 802 654 0638 Phone 86 10 85865569 Fax 86 10 85861829 Email TAC biotek com Website www biotek com Email infochina biotek com Website www biotekchina com cn BioTek Germany Service Center European Coordination Center BioTek Instruments SAS Bureau de liaison France Kocherwaldstrasse 34 D 74177 Bad Friedrichshall Germany 50 avenue d Alsace 68025 Colmar Cedex France Ph...

Page 12: ...0 Phone 81 0 3 5812 8109 Fax 81 0 3 5812 8115 Email info biotek uk com Website www biotek uk com Email infojapan biotek com Website www biotek com ja Revision History Rev Date Changes A 10 2013 First Issue B 5 2014 The manual was updated to add RAD Technology content throughout Syringe dispenser installation steps were updated to add the Import from USB calibration data feature Added references to...

Page 13: ...spenser provides microplate priming and dispensing and when equipped washing for ELISA fluorescence and chemiluminescence immunoassays cellular and agglutination assays l If the instrument has an IVD label it may be used for clinical and non clinical purposes including research and development If there is no such label the instrument may only be used for research and development and non clinical p...

Page 14: ...component previously listed BioTek will replace the cassette components free of charge Repackaging and Shipping If you need to ship the instrument to BioTek for service or repair contact BioTek for a Return Materials Authorization RMA number and use the original packing materials Other forms of commercially available packaging are not recommended and can void the warranty If the original packing m...

Page 15: ...to the microplate carrier or carrier track Lubricant on the carrier mechanism will attract dust and other particles which may obstruct the carrier path and cause the instrument to produce an error Warning Liquids Avoid spilling liquids on the instrument fluid seepage into internal components creates a potential for shock hazard or instrument damage If a spill occurs while a program is running abor...

Page 16: ...tions Do not expose the instrument to temperature extremes For proper operation ambient temperatures should remain within the range listed in the Specifications section Performance may be adversely affected if temperatures fluctuate above or below this range Storage temperature limits are broader Caution Sodium Hypochlorite Do not expose any part of the instrument to the recommended diluted sodium...

Page 17: ... electronic equipment WEEE or local ordinances Caution Warranty Failure to follow preventive maintenance protocols may void the warranty Caution Shipping Hardware All shipping hardware e g shipping bracket etc must be removed before operating the instrument and reinstalled before repackaging the instrument for shipment Caution Do not run the Peri pump without a cassette installed on the pump Cauti...

Page 18: ...ch case you may need to mitigate the interference Immunity The system has been type tested by an independent accredited testing laboratory and found to meet the requirements of EN 61326 1 and EN 61326 2 6 for Immunity Verification of compliance was conducted to the limits and methods of the following EN 61000 4 2 Electrostatic Discharge EN 61000 4 3 Radiated EM Fields EN 61000 4 4 Electrical Fast ...

Page 19: ...ostic IVD medical equipment Electromagnetic Interference and Susceptibility USA FCC CLASS A RADIO AND TELEVISION INTERFERENCE NOTE This equipment has been tested and found to comply with the limits for a Class A digital device pursuant to Part 15 of the FCC Rules These limits are designed to provide reasonable protection against harmful interference when the equipment is operated in a commercial e...

Page 20: ... Reglement sur le brouillage radioelectrique edicte par le ministere des Communications du Canada User Safety This device has been type tested by an independent laboratory and found to meet the requirements of the following l Underwriters Laboratories UL 61010 1 Safety requirements for electrical equipment for measurement control and laboratory use Part 1 general requirements l Canadian Standards ...

Page 21: ... accompanement AchtungsieheBegleitpapiere Atención vease los documentosincluidos Attenzione consultare la doc annessa Warning risk of electric shock Attention risque de choc électrique Gefährlicheelektrischeschlag Precaución riesgo de sacudidaeléctrica Attenzione rischiodiscossaelettrica Warning risk of crushing or pinching Attention risqued écrasement et pincement Warnen Gefahr des Zerquetschens ...

Page 22: ...onikgeräten Recogida selectiva de aparatos eléctricos y electrónicos Raccolta separata delle apparecchiature elettriche ed elettroniche Consult instructions for use Consulter la notice d emploi Gebrauchsanweisung beachten Consultar las instrucciones de uso Consultare le istruzioni per uso MultiFlo FX Multi Mode Dispenser ...

Page 23: ...ifications and includes important contact information Introducing the MultiFlo FX Dispenser 2 Features of the MultiFlo FX 3 MultiFlo FX Dispenser Comparison 7 BioStack Compatibility 10 Package Contents 12 Optional Accessories 13 Physical Specifications 18 Performance Specifications 21 Space Requirements 25 BioTek s Customer Resource Center 26 ...

Page 24: ...her or a single channel Random Access Dispenser RAD module The MutliFlo FX is available in multiple configurations to support numerous microplate geometries Device Component Description 1 Peri pump Dispenser Peristaltic 8 channel dispenser with entirely visible fluid path 2 Dispense arm Holds the Peri pump Tip Holder s the Syringe dispenser manifolds and the washer dispense manifold when applicabl...

Page 25: ...nal Peri pump dispenser required for RAD models See Plate Types Table on page 9 to review the plate types supported by each device Features of the MultiFlo FX l Supports all microplate based assays including ELISA fluorescence chemiluminescence RIA DNA probes and cellular assays l A variety of solutions including buffered saline and reagents can be dispensed l The intuitive onboard software allows...

Page 26: ...0 5 µL dispenses when using a 1 µL cassette l RAD Technology models support random access dispensing with several single channel cassette options including a bulk dispensing cassette 8 tubes to one chute with flow tracking to minimize turbulence in the wells l Autoclavable tubing steam temperatures and pressures of 121 C and 1 bar 750 mmHg is compatible with 70 ethyl or isopropyl alcohol and 0 5 s...

Page 27: ...r o Running Maintenance protocols o Running Qualification protocols o Special considerations when operating with the BioStack Microplate Stacker RAD Technology Features Random Access Dispensing or RAD models offer the most gentle dispensing possible to support sensitive cellular assays The single tube cassettes are ideal for reagent preservation hit picking or partial plate filling The 8 to 1 chan...

Page 28: ... gradually raises the tip chute as fluid is dispensed to the well to ensure the least disturbance possible See RAD Tip Tracking on page 156 cell monolayer is preserved MultiFlo FX Models Model Internal Peri pump Strip Washer RAD Technology MFX MFXW X MFXP X MFXPW X X MFXR X MFXPR X X The Syringe and secondary Peri pump dispensers are ordered separately Except RAD models include the external Peri p...

Page 29: ...volume See Shorten the Dispense Cassette Tubing on page 61 n RAD Technology offers the best tools for preserving cells and reagents n All MultiFlo FX devices are capable of dispensing up to 30 000 µL BioTek s recommended maximum dispense volumes are cited here Device Volume range µL well Precision Accuracy Approximate Dead volume Peri pump 0 5 and 1 3000 10 CV 1 µL well typical 3 CV 10 typical 3 1...

Page 30: ... speed to obtain the following processing times including the fastest flow and travel rates Some of these parameters are listed in the Parameters column of the table Only standard non RAD Peri pump cassettes are referenced here Device Plate Type Volume µL well Parameters Time in seconds Peri pump 5 µL 96 10 High flow rate 3 Peri pump 1 µL 384 1 High flow rate 6 1536 1 High flow rate 21 Syringe 8 t...

Page 31: ... Deep Well 24x16 44 08 N A 986 384 PCR 24x16 9 50 6 230 1536 Well 48x32 10 41 N A 250 1536 Flanged 48x32 10 26 N A 196 N A Plate washing not supported for this plate type Low Density Plates Peri pump non RAD Strip Washer Syringe Plate Type columns x rows Plate Ht mm Dispense Ht steps PP tubes well Washer Syringe tubes well 6 Well 3x2 20 20 465 4 4 12 Well 4x3 20 20 460 2 3 24 Well 6x4 20 50 452 2 ...

Page 32: ... height flange height 1 0 mm l Travel Height plate height flange height Plate Clearance n If the Dispense Height is greater than Travel Height the travel height is changed to match the dispense height BioStack Compatibility The MultiFlo FX is compatible with BioTek s BioStack Microplate Stacker The BioStack can rapidly transfer microplates one at a time to and from the instrument and includes l Re...

Page 33: ...perate with the MultiFlo FX refer to the BioStack Operator s Manual for instructions on configuring the MultiFlo FX to run with the BioStack To help you get started See Operating with the BioStack on page 124 If you are interested in purchasing the BioStack contact your local BioTek dealer for more information or visit our website at www biotek com BioTek Instruments Inc ...

Page 34: ...ng trough insert for Syringe dispensers 2 and Strip washer or RAD dispenser 7182044 Strip plate 12x1 98265 Screwdriver Phillips 98268 Accessory kit 1260004 Stylus for cleaning washer and Syringe manifold dispense tubes 2872304 10 cc syringe and tubing for Peri pump cassette maintenance 7210021 Shipping brackets 2 model dependent 1262010 1262011 1262115 Hex wrench 7 64 for removing shipping bracket...

Page 35: ... 7212042 Liquid Handling Control Software LHC2 BioStack Microplate Stacker and integration kit Biostack Installation Operational Performance Qualification IQ OQ PQ package 1260521 Peri pump Optional Accessories Secondary Peri pump assembly PN 7210010 Dispense cassettes and accessories Cassette Type Cassette Tips Replacement tubing kit Tubing extension kit 1 µL plastic tips 7170012 7172150 7170009 ...

Page 36: ...ey ship with plastic tips RAD Technology Cassettes PN 10 µL Single molded tip 1260015 5 µL Single molded tip 1260016 1 µL Single molded tip 1260017 10 µL Single steel tip 1260018 5 µL Single steel tip 1260019 1 µL Single steel tip 1260021 10 µL Large bore single molded tip 1260020 Bulk dispenser 8 tube to 1 chute 5 µL tubing molded tips 1260022 Random Access Dispenser RAD Accessory PN Cassette Cal...

Page 37: ...tubes well 1260534S Stylus for cleaning 8 16 tube dispense manifold tubes 2872304 Stylus for cleaning 32 tube LB dispense manifold tubes 7182095 Stylus for cleaning 32 tube SB dispense manifold tubes 7182102 Inline Filters 2 48705 Hex wrench 3 32 for removing syringe pumps 48570 Hex wrench 1 16 for removing magnets from dispense manifolds 48713 Spare tubing sets 2 1 dispenser autoclavable 7183006 ...

Page 38: ...Manifold 1262029 6 Well Accessory Kit 1260009 Dispense Manifold 1262052 Aspirate Manifold 1262053 12 Well Accessory Kit 1260010 Dispense Manifold 1262050 Aspirate Manifold 1262051 24 Well Accessory Kit 1260011 Dispense Manifold 1262048 Aspirate Manifold 1262049 48 Well Accessory Kit 1260012 Dispense Manifold 1262046 Aspirate Manifold 1262047 Magnetic Bead Assay Accessories Accessory PN Magnet Adap...

Page 39: ...Optional Accessories 17 Accessory PN 384 well Flat Magnet 7103017 384 well Ring Magnet 7102215 96 well Flat Magnet 7103016 96 well Ring Magnet 7102216 BioTek Instruments Inc ...

Page 40: ...icrowells Flat round V bottom Hardware Environmental User Interface 5 7 touch screen Power Supply The instrument uses two internal power supplies 24 volt 60 watt and 48 volt 60 watt These supplies are compatible with 100 240 V 50 60 Hz Dimensions W x D x H Approximately 17 19 x 11 75 x 8 0 inches 44 cm x 29 cm x 20 cm base model without strip washer external Peri pump or Syringe dispenser Weight 1...

Page 41: ...nifold types Manifold Type 8 Tube 2 x 8 channel non autoclavable manifold with replaceable stainless steel tubes to process 96 and 384 well plates 16 Tube 1 x 16 channel autoclavable manifold with replaceable stainless steel tubes to process 96 and 384 well plates 32 Tube 1 x 32 channel manifold cannot be autoclaved and does not support non factory tube replacement An inline 90 micron filter is in...

Page 42: ...cement syringe pump to dispense fluid and a vacuum pump to aspirate fluid Its dual action design uses separate plate type specific manifolds for aspirating and dispensing Manifold Type 96 384 Well 1 x 8 channel dispense and aspirate manifolds to process 96 and 384 well plates 48 Well 1 x 12 channel dispense manifold 2 tubes well and 1 x 6 channel aspirate manifold to process 48 well plates 24 Well...

Page 43: ... is valid at 0 5 1 2 3 30 000 µL Accuracy is measured gravimetrically when dispensing room temperature deionized water Specifications apply to volumes that are full unit increments for the cassette to which they apply For example the accuracy specification for a 10 µL cassette is valid at 10 20 30 30 000 µL Cassette Precision Accuracy 1 µL 10 CV 1 µL per well 10 1 µL per well 5 CV 2 µL per well 5 ...

Page 44: ...5 isopropyl alcohol solution The absorbance of the solution shall be read at 630 nm and 450 nm reference Accuracy is measured gravimetrically when dispensing room temperature deionized water Dispense Precision 8 Tube 2 CV when dispensing 100 µL well 5 CV precision at 20 µL well 5 CV precision at 40 µL well Tested in house to 4 0 CV 16 Tube 2 CV when dispensing 100 µL well 2 5 CV precision at 80 µL...

Page 45: ...ual Volume Evacuation Efficiency 96 Well Average residual of 2 0 µL per well after a 3 cycle wash when dispensing 300 µL per well using a bottom touching well aspiration and a solution of deionized water with 0 1 Tween 20 or equivalent buffer solution The aspirate height adjustment should be optimized for the plate prior to testing 6 12 24 48 Well Refer to the table below to determine the average ...

Page 46: ...formance Volume µL well 96 Well 3 0 CV 300 6 Well 5 0 CV 5560 12 Well 3 0 CV 2240 24 Well 3 0 CV 1120 48 Well 3 0 CV 560 384 Well 3 0 CV 80 Dispense Accuracy Washer dispense accuracy shall be 3 0 when deionized water with 0 1 Tween 20 and FD C 1 blue dye solution The weight of the fluid dispensed shall be measured gravimetrically Plate Type Volume µL well 96 Well 300 6 Well 1120 12 Well 2240 24 We...

Page 47: ... when installing a Strip washer and or Syringe dispenser to the back Similarly expect to add 8 20 cm to the depth when installing the RAD technology dispenser l Add 4 5 to the depth or width for fluid supply and waste bottles depending on placement l Plan on a height of approximately 12 31 cm when installing the washer the waste bottle is the tallest accessory or if the waste bottle resides elsewh...

Page 48: ...upport After an easy registration process you can access lots of useful information about your BioTek microplate instrumentation and software On the secure CRC website you can l Track orders l Access warranty information user manuals and software updates l Download technical and application information l Maintain equipment inventory product registration l Request service and technical support l Vi...

Page 49: ...spenser 44 Install the Secondary Peri Pump 46 Install RAD Technology Components 51 Install the Syringe Dispenser Component 62 Install Software Connect to Computer 71 Connect Power Cable 72 Define Instrument Settings 73 Verify Performance 79 Repacking the MultiFlo FX 82 Install the Shipping Hardware 83 Repacking the Secondary Peri pump 85 MultiFlo FX Repacking 86 Repacking the Syringe Dispenser 89 ...

Page 50: ...packing material for the carrier s inspection BioTek will arrange for repair or replacement of your instrument immediately before the shipping related claim is settled 1 Unpack the boxes containing the instrument and other equipment l MultiFlo FX Multi Mode Dispenser and accessories l Strip Washer and accessories l Dual Syringe Dispenser and accessories l Additional Peri pump Dispenser and accesso...

Page 51: ...g must lay flat to properly drain the priming trough The side bracket provided with the instrument supports this goal but it may not be needed in your lab depending on where you position the dispenser and its waste receptacle You will need l Philips head screwdriver small l Waste tubing 4 provided PN 7213010 l Possibly scissors or knife to cut tubing to desired length l Side bracket provided l Was...

Page 52: ...riming Trough 1 Thread the waste tubing under the bracket and onto the priming trough s spout Remove the bracket and or use water or alcohol to help the tube slide onto the spout 2 Snake the tubing around the instrument to the side bracket 3 Return the instrument to normal position Position Waste Container and Tubing When all the installation steps are completed 1 Place the waste container under t...

Page 53: ... installing the waste tubing for the priming trough place it upright on a level work surface to remove the shipping hardware Store the shipping brackets on the back of the instrument in slots provided for this purpose You will need l Allen or hex wrench taped to plate carrier The shipping hardware is slightly different than shown above for MultiFlo FX BioTek Instruments Inc ...

Page 54: ...ve shipping brackets 1 Remove the plate carrier shipping bracket use the Allen wrench to remove the two screws that hold it to the plate carrier Tilt the bracket downwards slightly and release it from the post 2 Remove the dispense arm shipping bracket use the Allen wrench to remove the two screws that hold the bracket to the instrument 3 Attach the shipping brackets to the back of the instrument ...

Page 55: ...est arm into the slot and use the Allen wrench to screw it in place Exception The dispense arm bracket for MFX and MFXP models does not fit into a slot Attach it with one screw so the short end projects out from the instrument You must first remove the large support bracket attached to the back panel that is used for an external Peri pump Insert the second screw next to the bracket Store the Allen...

Page 56: ...washer module Part Part Description Part Number Aspirate manifold install on dedicated aspirate arm Press the stainless steel tab on the front of the arm to install and to remove the manifold Install this manifold last after installing items on the dispense arm See Install Washer Manifolds on page 41 1260510 for 96 384 well shown Dispense manifold install on the dispense arm after installing eithe...

Page 57: ...Especially useful when performing magnetic bead assays See Special Procedure for Magnetic Bead Assays on page 151 01262 Tubing bracket to hold tubing in place 1262066 Vacuum Line Filter cut the vacuum line tubing and install the filter to prevent contaminates from clogging the vacuum pump 48146 See also Strip Washer Optional Accessories on page 15 for part numbers for plate type specific washer ma...

Page 58: ... back panel 2 Align the slots on the side of the washer module with the two posts near the base Simultaneously align the screw holes on the top of washer unit 3 Use the screws removed in the first step to attach the module to the instrument 4 Connect the serial cable from the washer module to its port on the back of the instrument back panel MultiFlo FX Multi Mode Dispenser ...

Page 59: ...D models Attach the bracket to the back of the instrument near the top center of the instrument slightly left of the aluminum mounting plate as you face the back panel You need a small Philips screwdriver 1 First remove the two screws you will use to attach the bracket Refer to the photo below to locate the screws on the back of the instrument 2 Align the bracket to the holes and use the screws to...

Page 60: ...es with check valves powered by a non autoclavable syringe pump 1 Locate the tubing with the Luer fitting on one end Gently screw the Luer fitting into its connection on the top of the supply bottle Finger tighten only 2 Attach the other end of the fluid supply tubing to the bottom port of the washer pump Make sure the arrow on the check valve points towards the pump 3 Connect the tube without fit...

Page 61: ...k valves and the rigid fixture of the waste tube in the center of the bracket As part of the setup Install Washer Manifolds on page 41 Install the Waste Bottle The washer collects waste fluid during aspiration The waste bottle is connected to the vacuum pump inside the washer module and the aspirate manifold to perform this function An inline vacuum filter not shown is optional but strongly recomm...

Page 62: ...per from the waste tubing set PN 1263001 2 Connect the tube without a fitting to the aspirate manifold 3 Install the inline vacuum filter on the tubing with a Luer fitting o Cut the tubing about two inches from the Luer fitting o insert the filter to reconnect the tubing 4 Gently screw on the Luer fitting to the port labeled Vacuum 5 If applicable connect the waste sensor cable to the Waste Sensor...

Page 63: ... distinct manifolds the dispense manifold resides on the dispense arm along with the Peri pump cassette s Tip Holder and the dual Syringe dispensers manifolds if equipped The aspirate manifold resides on its own dedicated aspirate arm Important Either the dual Syringe dispenser manifolds or the spacer block must be installed on the dispense arm before the washer dispense manifold can be installed ...

Page 64: ...ng closest to the instrument Typically magnets hold them securely 6 With its supply tube closest to the instrument guide the dispense manifold onto the dispense arm Its magnet will hold it securely Alternatively especially when running magnetic bead assays remove the magnet and use the thumb screws provided Special Procedure for Magnetic Bead Assays on page 151 7 Lift the aspirate arm to its highe...

Page 65: ...ifolds on the dispense arm instead of the Spacer block if applicable To remove the washer manifolds Reverse the order of the installation steps i e first remove the aspirate manifold press the release tab Then remove the dispense manifolds BioTek Instruments Inc ...

Page 66: ...h the Peri pump cover panel l Dispense cassette l Fluid supply vessel l Optional Prime trough insert Attach the Peri pump Cover Panel Optionally if you are not installing an external or secondary Peri pump install the cover panel provided to conceal the primary Peri pump Install the cover panel using the two hex screws and the Allen wrench shipped with the MultiFlo FX MultiFlo FX Multi Mode Dispen...

Page 67: ...Set Up the Peri pump Dispenser 45 MultiFlo FX with closed Peri pump cover panel BioTek Instruments Inc ...

Page 68: ...ith its own accessories and it is required for RAD technology Inspect and unpack the shipping container and follow these installation instructions if applicable Otherwise skip this part of the installation process The secondary Peri pump is designed to sit on top of the MultiFlo FX above and slightly right of the internal Peri pump MultiFlo FX Multi Mode Dispenser ...

Page 69: ...ace 2 l Also note the screwdriver Allen hex wrench and shoulder screw shipped with the accessories You will need l Second Peri pump Kit components shown above l Philips head screwdriver small l Small flat screwdriver Install the additional Peri pump MultiFlo FX ONLY Change the bracket on the front of the Secondary Peri pump unit l Remove the black plastic tab and replace it with the stainless stee...

Page 70: ...cket 3 Place the second Peri pump unit on top of the dispenser 4 Position the dispenser so the screw hole in the small tab or stainless steel bracket attached to its front aligns with the right hand screw hole on the primary Peri pump or cover plate 5 Use the Philips screwdriver to fully tighten the shoulder screw to ensure the unit is secured 6 On the back of the instrument secure the Peri pump u...

Page 71: ... lit It s a tight space and the second cassette bracket fits snugly onto the original bracket 1 First remove the four screws and washers from the additional cassette bracket using the small Allen wrench supplied in the kit 2 Slide the plate carrier to the left out of the way Lower the dispense arm to a workable position 3 Use the text on the front of the additional cassette bracket to orient it co...

Page 72: ... on the back of the instrument and to the port on the back of the Peri pump unit Use the small flat screwdriver to secure the cable n If you are installing a Syringe dispenser also you may find it easier to complete that installation before connecting the cable turning on the instrument and installing the dispense cassettes MultiFlo FX Multi Mode Dispenser ...

Page 73: ...gy uses the secondary or external Peri pump dispenser a tubing bracket and a dedicated dispense head After removing and storing the shipping hardware install these components l Install the Secondary Peri Pump on page 46 l Install the Tubing Bracket and RAD Module below l RAD Plug in the cables on page 53 l Install RAD Dispense Head on page 53 l Install RAD Technology Cassette on page 58 Install th...

Page 74: ...the instrument slightly left of the aluminum mounting plate as you face the back panel You need a small Philips screwdriver 1 First remove the two screws you will use to attach the bracket Refer to the photo below to locate the screws on the back of the instrument 2 Align the bracket to the holes and use the screws to attach it Attach the RAD Module to the Tubing Bracket View of inside the tubing ...

Page 75: ...tive ports l RAD module to Washer Module port l Peri pump to Peri Pump Dispenser port Install RAD Dispense Head The RAD dispense head resides on the dedicated Aspriate manifold RAD arm Its ribbon cable plugs into the RAD module installed in earlier step 1 Align the two pins on the dispense head with the two holes on the RAD arm and press the Release Tab on the front of the arm to install the dispe...

Page 76: ...o the RAD module installed on the tubing bracket Next install the RAD cassette Install RAD Technology Cassette on page 58 Dispense Cassette Diagram 1 Tip Holder 2 Center Holder 3 Tube Tensioner 4 Tubing Organizer 5 Tip Guard remove before use MultiFlo FX Multi Mode Dispenser ...

Page 77: ... grooves on the underside of the pump 3 Tube Tensioner The transparent 5 mm scale on its front surface identifies the tube tensioner It has internal screws for stretching the tubing one for each tube The tube tensioner s scale is useful when calibrating the cassette 4 Tube Organizer At the opposite end of the cassette from the tip holder the tube organizer holds the 8 tubes together for inserting ...

Page 78: ... Secondary pump s Tip Holder in the right bracket o Release the pump s stainless steel plate Use your right hand to release the spring loaded cassette latch on the right side of the caution symbol on the pump and use your left hand to lift the stainless steel plate up and out The BioTek logo on the Tip Holder faces the Peri pump The logo on the Center Holder faces down away from the pump Note See ...

Page 79: ...lide the Center Holder into its slot on the underside of the pump A tab on the back of the center holder fits into a notch on the pump The label on the center holder faces down 3 Align the Tube Tensioner with the stainless steel plate as it wraps around and up against the pump Be sure the knobs on top of the tensioner fit correctly into the grooves in the stainless steel plate as you move both par...

Page 80: ... only They are not designed for use on an internal Peri pump Follow instructions to Install the Dispense Cassette on page 56 skipping the first step Magnets on the dispense head hold the cassette tip s in place Single tube RAD cassette 8 tube to 1 Bulk RAD cassette Align the two pins with the holes above and below the magnets on the dispense head n You must create a protocol to successfully use th...

Page 81: ...2043 for the Peri pump dispenser holds approximately 12 mL 1 per dispenser l PN 7182044 2 for the Syringe dispensers 1 for the washer holds approximately 6 5 mL n Without the prime trough insert the priming trough empties into the regular waste bottle Important Remove the priming trough inserts when using the 8 to 1 RAD cassette The chute will collide with both the RAD priming cup and the Syringe ...

Page 82: ...ette Tubing on the facing page The reservoir holder has two components a bracket and a strap 1 Clip the bracket onto the front of the Peri pump When properly installed the bracket hides the hazard label 2 Place the supply vessel reservoir on the bracket and wrap the strap around both the vessel and the bracket fit the strap s molded balls into the round slots Depending on the size of the vessel it...

Page 83: ...determine where to cut the tubing l Cut one tube at a time using a razor blade Exacto knife or sharp scissors make as clean a cut as possible l Cut the tubing as evenly as possible limiting the difference between the tubes to less than 1 4 6 35 mm not applicable to RAD single channel cassette To may make this job easier use the cassette s packaging to hold the cassette parts keeping them out of yo...

Page 84: ...t applicable l 1 Syringe Dispenser module autoclavable l 2 interface cable l 3 tubing bracket l 4 priming trough inserts l 5 dispense manifolds l 6 shelf l 7 supply bottles and tubing with check valves l Not shown thumbscrews 2 and a USB flash drive Review the placement options for the Syringe dispenser and follow the applicable instructions Syringe Dispenser Placement Options on the facing page M...

Page 85: ...sher module The second option is on top of the instrument itself When installing the Syringe dispenser on top placement of the tubing bracket varies slightly depending on how many other dispensers are installed Follow the installation instructions for your preferred placement of the dispenser Install the Syringe Dispenser on Back Shelf on the next page or on top of the washer Install the Syringe D...

Page 86: ...nd use the washer as a shelf if applicable First reposition the shelf s feet 1 Remove the shelf shipped with the Syringe dispenser from its plastic bag PN 7212038 2 Unscrew the rubber feet that are shipped in the wrong position 3 Install the feet in the correct position screw the feet into the two outer holes on the bottom of the shelf The third hole is for BioStack integration 4 Remove the two sc...

Page 87: ...t over the screw and tighten both screws 7 Put the syringe pump module on the shelf or the washer module 8 Plug the serial cable into the back panel of the MultiFlo FX in the port labeled Syringe Pump Dispenser Plug the other end into the syringe pump unit Next steps l Update the instrument to use the Syringe Dispenser on page 74 l Install Tubing and Manifolds for Syringe Dispenser on page 68 Inst...

Page 88: ...the support bracket from the back of the instrument by releasing the two screws that hold it in place 2 Align the bracket with the screw holes on the back of the instrument so that the foot holds sit above the instrument or second Peri pump s top 3 Install the tubing bracket in the appropriate location using the two black capped screws Tubing Bracket When installing the Syringe dispenser on top of...

Page 89: ...nly no Peri pump Remove the two screws in the filler plate on the front of the instrument and replace them with the tubing bracket 4 Plug the 26 pin high density cable into the back panel of the MultiFlo FX in the port labeled Syringe Pump Dispenser Plug the other end into the syringe pump unit Next steps l Update the instrument to use the Syringe Dispenser on page 74 l Install Tubing and Manifold...

Page 90: ...mum pump performance l Make sure the tubing is not crimped during installation Perform these steps twice first for Syringe A and then for Syringe B 1 First install the inline filter for 32 tube dispensers if applicable as described on page 71 Locate the tubing with a Luer fitting on one end Gently screw the Luer fitting into the top of the supply bottle Finger tighten only 2 Attach the other end o...

Page 91: ...ld that has two manifolds in one block install Syringe A s manifold first Syringe B s manifold must be installed after Syringe A s 5 Connect the supply tube to the dispense manifold n Images of syringes on the instrument labeled A and B indicate the placement of each dispenser s manifold A slides on first then B n Special procedure required for magnetic bead assays A magnet holds the two manifolds...

Page 92: ...ily move the pump unit to get access to the tubing bracket n When the Syringe in installed on top of the washer use the washer s tubing bracket instead 7 Repeat the tubing and manifold installation for Syringe B 8 Optional Insert the thumbscrews into the posts that hold the manifolds Magnets keep the manifolds attached to the dispense arm The thumbscrews are provided in case the magnets interfere ...

Page 93: ...nt toward the pump from the supply vessel and towards the dispense manifold from the pump Install Inline Filter for 32 Tube Dispensers BioTek ships two 90 micron inline filters with the 32 tube dispense manifolds to reduce the chances of clogging the dispense tubes It is especially important for the SB small bore models To install the filters 1 Locate and layout the length of tubing that goes betw...

Page 94: ...CD n The Home screen must be displayed for the LHC to communicate with the instrument Technical Note Only one of the two communication ports COM port on the instrument can be used at a time They cannot be used simultaneously You can use USB to connect the MultiFlo FX to the computer or the RS232 serial port to connect to a BioStack or similar robotic device But you cannot use both ports simultaneo...

Page 95: ...on the main page and if required select the MultiFlo FX 2 Specify the COM Port used to connect the MultiFlo FX to the computer use the drop down list to select the port and click Test Communication o Pass proceed to the next step o Fail check the Com Port setting See About Com Ports in the LHC Help 3 In the Target Instrument Settings dialog that opens click Get actual settings now and click OK Ena...

Page 96: ...MultiFlo FX define the correct COM port 3 Select Tools Instrument Utilities Syringe Dispenser 4 Under Syringe Dispenser Assembly specify the type of pump and the installed manifold type and click Send to download the data to the instrument 5 In the Calibration Data section enter the data points on the label in the corresponding fields including the Serial Number or plug the USB flash 1 Connect the...

Page 97: ...ntered When importing data from the USB flash drive confirm the serial number on the USB matches the one on the Syringe unit Change the Syringe Dispenser Manifold Changing the Syringe manifold requires two steps l Physically changing the Syringe dispenser manifold See Install the Syringe Dispenser Component on page 62 l Updating the instrument s manifold setting as described below After physically...

Page 98: ...either internal or external it is called Primary LHC Touch screen 1 Select Tools Instrument Utilities 2 Select the Peri pump tab 3 Select the correct button for the Cassette Type installed for the primary or secondary Peri pump if applicable 4 Click Send to update the instrument 1 From the home screen select Instrument Peri pump 2 Select the Cassette Type for each dispenser Primary and Secondary a...

Page 99: ... to set the 24 hour clock Touch the number field to put it in edit mode l Date Touch today s date on the calendar to select it Press the previous or next arrows up or down to scroll to the correct month 4 Press Save 5 Restart the instrument Remember to update the time in accordance with annual time shifts like Daylight Savings Time if applicable Note The sort order of the protocol list on the Home...

Page 100: ...e g Template LHC 5 Select Tools Preferences New Protocol 6 Select the button for Protocol selected below to use as a template 7 Click selected and select the protocol you created as a template Define startup behavior 8 After completing the steps above select the Startup Options tab 9 Select the button for New Protocol 10 Click OK to save your new preferences MultiFlo FX Multi Mode Dispenser ...

Page 101: ...error message is displayed l Fail an error message is displayed If this happens note the error code and refer to Troubleshooting on page 272 to determine its cause If the problem is something you can fix turn off the instrument fix the problem and then turn the instrument back on Otherwise contact BioTek s Technical Assistance Center n Note An instrument qualification package PN 1260521 for the Mu...

Page 102: ...utton Verify both a standard cassette and a RAD technology cassette if applicable If leaks were detected tighten all tube fittings and re run the program If leaks are still detected contact BioTek s Technical Assistance Center Verify the Syringe Dispensers 1 Fill the supply bottles with approximately one liter of deionized water one for each syringe pump LHC Touch screen 1 Select File Open 2 Open ...

Page 103: ...tiFlo FX and Maintenance Protocols folders and then open W DAY_RINSE LHC You may need to reset the Com Port 3 When ready click the Run button to prime the tubing and manifold with deionized water 4 When finished close the program 1 Select Maintenance at the Home screen 2 Select W Day_Rinse and press Start 3 When the protocol is completed press the Home button If leaks were detected tighten all tub...

Page 104: ...1 Remove the Peri pump dispense cassette and the Syringe dispenser and washer manifolds if applicable 2 Uninstall and repack separately the Syringe pump and its accessories and the strip washer and its accessories if applicable 3 Slide the plate carrier into position next to the dispense arm It will be secured in place by one of the shipping brackets View the illustrations provided l Repacking Sec...

Page 105: ... You will need l Allen or hex wrench provided with the instrument l The shipping brackets removed from their storage slots on the back of the instrument Reverse the process of Removing the Shipping Hardware The shipping hardware is slightly different than shown above for MultiFlo FX without a washer or RAD technology BioTek Instruments Inc ...

Page 106: ...r as it will go and raise the dispense arm Does not apply to MFX and MFXP models 3 Secure the dispense arm with its shipping bracket aligning it with its screw holes and use the Allen wrench to install the two screws that hold it to the instrument 4 Slide the plate carrier to the left positioning it below the dispense arm 5 Insert the plate carrier shipping bracket rest the bracket on its post and...

Page 107: ...not shipped Uninstall the Secondary Peri pump 1 Reverse the installation procedure Install the Secondary Peri Pump on page 46 to remove the Secondary Peri pump from the top of the MultiFlo FX 2 Wrap a rubber band around the pump itself 3 Place the module in the plastic bag and put it in its shipping box RAD Technology Components 1 Put the RAD components into their respective plastic bags and then ...

Page 108: ...ls to secure the items in place 3 Tape the box closed This box will fit into the box with the instrument MultiFlo FX Repacking 1 Reverse the steps described to Remove the Shipping Hardware First remove the two shipping brackets from the back of the instrument then install them 2 Put a rubber band around the Peri pump MultiFlo FX Multi Mode Dispenser ...

Page 109: ...g hardware installed sit the instrument in the foam edged box bottom 4 Place the end caps on either side and put all items inside the interior shipping box 5 Place accessory boxes inside interior box 6 Tape the box closed BioTek Instruments Inc ...

Page 110: ... eight foam corner blocks on the interior box and put it inside the outer box 8 Tape the outer box closed 9 Write RMA in large clear letters on the outer box 10 Include the RMA number in the address label MultiFlo FX Multi Mode Dispenser ...

Page 111: ...aring the instrument for shipping and reversing the installation steps pack the Syringe dispenser as shown here Fill the foam tray with the Syringe dispenser accessories and place the tray on top of the inner foam box containing the pump and supply bottles BioTek Instruments Inc ...

Page 112: ...istance Center for guidance Uninstall the strip washer 1 Reverse the installation procedure as described on page 35 to detach the strip washer from the MultiFlo FX 2 Separate the manifolds from their tubing and allow all fluid vessels to completely dry 3 Pack the washer module and its accessories in the shipping container 4 Repack the cardboard and foam shipping materials to secure the items in pl...

Page 113: ...29 Plate Types and Processing Patterns 133 Handling Special Plates and Mini tubes 134 Dispense Processing Patterns 138 Washer Operation 141 Cell Wash 142 Biomagnetic Separation Magnetic Bead Assays 145 Peri pump Peristaltic Dispenser 152 Peri pump How it works 153 Peri pump Dispense Step 155 Require a Specific Peri pump Cassette 158 Peri pump Advanced Settings 159 Dual Syringe Dispenser 160 Syring...

Page 114: ...Wash Dispense Protocols Touch a protocol on the Home screen to run it All saved predefined protocols appear on the Home screen Numbers Touch any field that requires a numeric value to open a field sensitive number pad Use the number pad to specify volume times and other values To enter negative numbers use the down decrement arrow Pick Lists touch a field with a down arrow to open a pick list to s...

Page 115: ...n Green buttons make the MultiFlo FX perform Quick Prime At the Home screen select Quick Device Prime Priming removes air bubbles from the tubing ensuring optimal performance Fluid is dispensed to the priming trough You can use Quick Prime to comply with the daily maintenance recommendations Quick Prime works best immediately prior to running a protocol to correct for evaporation i e to normalize ...

Page 116: ...setting matches the currently installed cassette See Change the cassette type setting on page 76 Refer to instructions for physically changing the cassette Install the Dispense Cassette on page 56 n 0 5 µL dispensing is not an option via the Quick menu You must define a protocol to dispense half microliter 500 nL l Press Volume to define the per well dispense volume in microliters µL Depending on ...

Page 117: ...and press Start to wash the plate Learn more l Create a protocol to save a record of the wash parameters to confidently repeat the wash procedure and or to modify wash parameters to improve performance l See Wash Parameters Table on page 108 to review options available in a wash protocol l See About the Wash Step Add protocols to the Home screen The MultiFlo FX puts all saved protocols on the Home...

Page 118: ...t Next Other from the Home screen 3 Press Transfer Protocols 4 Choose the File Source USB to copy protocols from it to the washer Conversely choose Internal to copy protocols to the USB flash drive to share them with another instrument 5 Optionally apply the Lock files option to prevent users from changing the protocols 6 Select files individually by highlighting them touch or transfer all protoco...

Page 119: ...Touch Screen Basics 97 l Ongoing operation Check your instrument setup for any unintended changes are the serial cables properly connected to the back of the MultiFlo FX BioTek Instruments Inc ...

Page 120: ...ssuring optimal performance l Check the external tubing connections for kinks and clogs l Put microplates on the carrier with well A1 in the left rear corner as you face the instrument and firmly seat the plate in the carrier l To more quickly dispense to 384 and 1536 well plates use the Instrument Utilities to change the Dispense Pattern to Row If precision is more important than speed keep the p...

Page 121: ...y prime the Syringe dispenser to ensure precision and accuracy increase the number of prime cycles to adequately remove all air bubbles from the tubing l Use the priming trough insert to capture expensive reagents for reuse Each Syringe dispenser s insert can hold up to 6 mL Recommended prime volumes for the Strip Washer The predefined maintenance protocol for the strip washer fully primes the sys...

Page 122: ...nsing protein solutions first prime the tubing with a buffered saline solution to remove any traces of water in the tubing then prime with the dispense fluid 2 Press the Prime button until fluid flows into the priming trough and all visible air bubbles have been removed At the end of the day Purge the tubing to reclaim the dispense fluid then Prime the tubing to flush it clean 1 Press the Purge bu...

Page 123: ...ime the quick prime is run The approximate dead volume for each Syringe dispenser system is 12 mL Generally three times the dead volume completely primes the system When using precious fluids e g expensive reagents you can change the prime parameters reduce the volume or number of cycles specified in the predefined protocols or create your own protocols Use the priming trough inserts to capture ex...

Page 124: ...es the wells 1 Edit the Wash step or stand alone Aspirate step Protocols Edit 2 Select Advanced Options for Aspirate and enable Perform Secondary Aspirate 3 Use the Align tool to define a crosswise aspirate position 4 Apply a second aspiration to the Final Aspirate also for best evacuation results For example you may want to adjust the X or Y axes to get to hard to reach areas of certain types of ...

Page 125: ...sert a step Copy a similar protocol and then edit it to meet different requirements or to test different parameters When editing a protocol cut and paste steps to move them to the correct position 6 Continue adding or editing steps as needed 7 Press Save to save the protocol All saved protocols are put on the Home screen for easy retrieval Optionally do a test run to verify the protocol performs a...

Page 126: ...is dispensed for each cassette type Cassette Type 1 μL 5 μL 10 μL μL sec tube Low 50 120 140 Medium 60 140 160 High 64 160 180 RAD technology Single channel cassette flow rates match the standard cassettes and the 8 to 1 cassette matches the 10 µL flow rates When dispensing 0 5 µL with the 1 µL cassette the average flow rate values are 1 μL Cassette 0 5 μL Dispense μL sec tube Low 52 Medium 54 Hig...

Page 127: ...ses the flow direction Volume 1 3000 µL 300 Duration 1 300 seconds 3 Note Half microliter 0 5 µL dispensing requires a 1 µL cassette The Cassette Requirement Mode behavior is implemented whenever a 0 5 µL volume is requested Make sure a 1 µL cassette is installed and the Cassette Type setting matches it Only a 0 5 µL dispense volume can be requested not 1 5 or 2 5 µL for example Perform two dispen...

Page 128: ...ense accuracy Begin by setting the delay to 500 msec Experiment with different settings to determine the optimal value for your fluid Submerge tips To soak dispense tubes in the priming fluid for a specified duration for cleaning or maintenance purposes If yes set duration up to 24 hours in minutes See Syringe Prime Step on page 161 to learn more 0 Dispense Flow rate Rates 1 5 are dependent on the...

Page 129: ...s are volume and plate type dependent For example rate 1 must be used when dispensing between 10 19 µL to a 96 well plate When dispensing 20 49 µL to a 96 well plate you can use rates 1 or 2 And when dispensing 50 59 µL to a 96 well plate you can use rates 1 2 or 3 And so on as shown in these tables 96 well plate 16 Tube 8 Tube Volume µL Rate µL sec well 10 19 1 450 140 20 49 1 2 600 209 50 59 1 3...

Page 130: ...ispense Rate 5 s minimum Manifold Flow Minimum Volume Rate 1 Rate 2 Rate 3 Rate 4 Rate 5 6 Well Flow 560 836 1116 1400 1680 Min Vol 40 80 200 240 320 12 Well Flow 420 627 837 1050 1260 Min Vol 30 60 140 160 220 24 Well Flow 280 418 558 700 840 Min Vol 20 40 100 120 160 48 Well Flow 279 372 560 600 800 Min Vol 40 80 200 240 320 Wash Parameters Table See Protocol Parameters Tables on page 103 for Pe...

Page 131: ...wells Define a delay between 0 5000 ms Increasing the delay may improve evacuation of the wells 0 Positioning X and Y horizontal axes Z height vertical axis can be adjusted to improve performance The default Z axis setting is plate dependent See Z Axis Default Dispense and Aspirate Heights Z 91 for 96 well plates X Y 0 Secondary aspirate Also called Crosswise Aspiration First the wells are aspirat...

Page 132: ...reate cleaning turbulence at the specified height The manifold descends to aspirate again and ends with a final dispense to fill the wells No Between cycles Shake To mix the contents of the plate No Duration From one second to one hour 5 sec Intensity Intensity Variable cycles through the other levels of intensity Slow Medium Fast Medium Soak Delays wash for the duration to allow fluids in the pla...

Page 133: ... plate are aspirated then dispensed to and so on Washer Flow Rates The default setting for flow rates is 5 in the middle of the range Rates increase from the slowest to fastest 11 Slower rates are recommended for viscous fluids Rate 1 is designed for cell wash assays causing the least disturbance in the wells Rate µL tube second Cell Wash 1 123 Cell Wash 2 140 3 210 4 252 Default 5 279 6 348 7 422...

Page 134: ...20 120 80 60 60 6 40 150 80 80 60 7 55 225 100 110 70 8 50 225 140 105 70 9 60 225 160 120 100 10 100 400 260 200 105 11 110 450 290 220 150 Aspirate Travel Rates Rate mm sec 0 CW 1 0 for 6 12 24 well plates 1 4 1 1 Slowest 1 CW 4 1 2 5 0 1 2 CW 5 0 3 7 3 1 Default rate 3 CW 7 3 4 9 4 1 4 CW 9 4 5 9 4 2 Fastest 6 CW 14 7 Recommended rate for Cell Wash protocols MultiFlo FX Multi Mode Dispenser ...

Page 135: ...pense Wash and Dispense Step option Also called Tip Priming About the Pre Dispense Options Enable the Pre dispense options to condition or normalize the dispense tubes before dispensing fluid This is recommended for precise dispensing to mitigate evaporation or any potential fluid imbalance at the ends of the tubes The pre dispense options are not intended to fully prime the tubing for example whe...

Page 136: ...g turbulence at the specified height The manifold descends to aspirate again and ends with a final dispense to fill the wells Designed for strongly bound molecules in assays that require vigorous washing e g ELISA adding a bottom wash to the protocol has shown reduced background noise without increasing the number of wash cycles Adjust the Z axis setting to position the manifold deep in the well t...

Page 137: ... Select Protocols at the home screen 2 Highlight the protocol and press Edit 3 Press the Info button on the step definition screen and touch the Plate Type field to change the plate type 4 Press OK to save the change Press Save at the step definition screen to save the protocol Rename a protocol There are two ways to rename a protocol edit the name or make a copy and rename the copy The MultiFlo F...

Page 138: ...Magnetic Bead Assays Set the shake intensity to Slow when using a magnet on the plate carrier l Soak allows buffer dispense fluid to remain in wells for the specified duration i e processing is delayed l Duration enter values to stop processing the plate for up to 1 hour 59 minutes 59 seconds n Soak is not the same as submerge the tips You must define a prime step or use the AutoPrime feature to s...

Page 139: ...delay which can be manually overridden when necessary l or for more precise control a timer to launch a protocol step This timer activated delay lets you apply a uniform processing time to each plate in a multi plate run Delay Type Indefinite to delay the procedure until the operator manually restarts the run A message box will open with a Continue button Any Prompt text will be displayed on scree...

Page 140: ...r every plate to ensure for example that reagent remains in every plate for a minimum or uniform time period This is especially beneficial when designing a multi step BioStack protocol that is run with varying numbers of plates in the stack If the timer is not used in this scenario assay performance could be significantly different in a run of 30 plates versus a 10 plate run n Important if the tim...

Page 141: ...uch longer than the duration of the Peri pump dispense operation Nested Delays The timer activated delay can be nested within another time controlled series of protocol steps This may be necessary to specify multiple processing start times for a complex series of steps including loops and BioStack steps This example uses a BioStack rack of plates The process requires that each plate in the rack be...

Page 142: ...stomize the Predefined Protocols below 1 Touch a protocol on the Home or Maintenance screen to select it 2 Press Start to run it LHC Users Only Customize the Predefined Protocols BioTek provides predefined protocols for maintenance routines and instrument qualification tests You can quickly customize the protocols for regular use The LHC keeps track of the last used COM port for an instrument type...

Page 143: ...Y_ RINSE_A B Two step protocol to flush tubing for both syringe pumps A B 40 mL per syringe Similar protocols with one step for each individual syringe are also provided S DAY_RINSE_A or_B P UL_CASS_ RINSE Where matches the cassette type in use and RAD refers to RAD technology cassettes Simple one step protocol to flush Peri pump tubing P2 protocols are for the secondary Peri pump W OVERNIGHT_ LOO...

Page 144: ...ol for 96 well wash manifold QC_96_EVAC_TEST Evacuation efficiency test protocol for 96 well wash manifold QC_SA 1536_ DISP_TEST Dispense precision test protocol for 32 tube Syringe A manifold QC_SB 1536_ DISP_TEST Dispense precision test protocol for 32 tube Syringe B manifold QC_P 1536_DISP_ TEST Dispense precision test protocol for Peri pump 1 µL cassette and 1536 well plate P2 protocols are fo...

Page 145: ...sition the manifold tubes for dispensing and aspirating to Corning Costar flat bottomed and round bottomed wells W COSTAR_ ROUND W NUNC_384 Standard wash protocols modified to best position the manifold tubes for dispensing and aspirating to Nunc flat bottomed wells round bottomed wells and 384 well plates W NUNC_FLAT W NUNC_ ROUND RAD Technology Peri pump Cassettes P_RAD 96 100 µL well Peri pump ...

Page 146: ...control them with the LHC l Design protocols that integrate BioStack controls with MultiFlo FX steps LHC protocols must contain a BioStack loop l In the LHC select Help Tutorials click Sections in the toolbar for a drop down menu select Controlling the Bio Stack with LHC It only takes a couple minutes to complete this interactive demo It is a great way to learn about the special BioStack features ...

Page 147: ...he BioStack s gripper before turning it on Robotics integrators CAD drawings of the physical dimensions of the MultiFlo FX are available upon request Contact BioTek customer service Technical Note Only one of the two communication ports COM port on the instrument can be used at a time They cannot be used simultaneously You can use USB to connect the MultiFlo FX to the computer or the RS232 serial ...

Page 148: ...tack A Restack step must be outside the BioStack step container 4 When all the protocol steps are defined click Validate to make sure the protocol is correctly designed 5 Save the protocol 6 At runtime except when using plates with lids you can define the Number of plates to process in the Run Info section of the run screen enter a specific number or select the Entire input stack for processing Ho...

Page 149: ...ly available when using the BioStack and plates without lids And the Nth Plate step must be put inside a BioStack step container n To precisely time the events in a BioStack run also consider the Delay option Timer Activated Delay on page 118 About the Nth Plate Read About the Nth Plate step if you haven t already done so See Also Advanced Plate Sequencing with the BioStack for more details and to...

Page 150: ... Nth Plate start and end tags 5 Click Validate to make sure the protocol syntax is correct Add a Delay step to the protocol Nth Plate steps to allow time during processing to put a special plate like a priming plate on the instrument or to do another manual task About the Nth Plate Step n The Nth Plate feature is only available when using the BioStack and plates without lids And the Nth Plate step...

Page 151: ... to determine which plates will be handled in a special way Any steps that precede an Nth Plate block will be performed on all plates including the Nth designated plates Multiple Nth Plate blocks can be defined in a protocol to sequentially perform a series of varying actions on a specified grouping of plates Advanced Plate Sequencing with the BioStack Read About the Nth Plate Step on the previous...

Page 152: ...3 300 µL 100 µL B 4 100 µL 200 µL 5 200 µL 200 µL 6 300 µL 200 µL C 7 100 µL 300 µL 8 200 µL 300 µL 9 300 µL 300 µL Use an Nth Plate step to perform maintenance Certain fluids like BSA can clog the liquid handler s dispense tubes so flushing the tubes after dispensing to a certain number of plates is recommended for optimal performance When using the BioStack you can use the Nth Plate step to add ...

Page 153: ...quired for an assay Don t forget to define the Number of plates to process for all the BioStack loops in the protocol Enter the number of plates at runtime or when defining the protocol In this example we create groups of like processed plates The stack will contain in this sequence 5 plates with 5 cycles 5 plates with 4 cycles 5 plates with 3 cycles and 5 plates with 2 cycles The plates in the Bi...

Page 154: ...ons Examples Nth Plate To generate mixed groups of dilutions in the stack of plates as depicted in the table below three Nth Plate steps are used The entire stack can be processed this way 1st Nth Plate Every 3rd plate starting with 1 2nd Nth Plate Every 3rd plate starting with 2 3rd Nth Plate Every 3rd plate starting with 3 Plate Volume 1 100 µL 2 200 µL 3 300 µL 4 100 µL 5 200 µL 6 300 µL 7 100 ...

Page 155: ...ultiFlo FX can process several plate types The default parameters for wash and dispense steps represent the optimal positioning of the hardware for the plate type And the aspirate and dispense heights and horizontal positions can be adjusted when necessary for special situations and to optimize assay performance l Review the Plate Types Table on page 9 for a listing of supported plates and their g...

Page 156: ...her the Syringe or Peri pump i e run multiple protocols on a plate when the assay requires using more than one device To achieve the best dispense performance when processing 1536 well flanged plates the MultiFlo FX s default protocol settings specify a low dispense height to position the dispense tubes as close as possible to the wells When the dispense arm holds multiple dispense manifolds the m...

Page 157: ...ions For some plates multiple tubes can dispense to a single well Conversely dispense tubes can and sometimes must be removed from the supply vessel or from the cassette to prevent them from missing the wells Testing at BioTek found the following capabilities Plate Type Columns Rows Tubes per Well Tubes removed 6 Well 3 x 2 3 or 4 4 and 5 or 0 12 Well 4 x 3 2 3 and 6 24 Well 6 x 4 2 0 48 Well 8 x ...

Page 158: ...luster Tubes PN 4410 4411 Special 24 Vial Rack To dispense to the mini tubes the special plate carrier must be installed to accommodate the extra height You can also use the carrier for other oversized vessels that may not otherwise be addressed by the dispensers by increasing the Plate Clearance setting The maximum height supported with the combination of special carrier and Plate Clearance is ab...

Page 159: ...e dispensers to fill the mini tubes n Note Position adjustments in the Y axis are prohibited with the special plate carrier for mini tubes Dispensing to Mini Tubes Unlike most dispensing with the MultiFlo FX the fluid stream does not hit the center of the well or tube when dispensing to the mini tubes using the special plate carrier This image illustrates where the fluid stream is positioned depen...

Page 160: ...With this feature you can dispense to odd columns in one dispense step change fluids or use another dispenser and dispense to even columns in another dispense step Rows 8 channel dispensers offer another way to control the dispense pattern to high density plates See 8 Channel Dispense Patterns 32 Tube Syringe Dispenser Manifolds One 32 tube dispense manifold addresses one column at a time When bot...

Page 161: ... Well Plate Rows You can skip one of the two rows sections When an 8 channel dispenser addresses a 384 well plate it first dispenses to odd numbered rows and then dispenses to even numbered rows With this feature you can dispense to odd rows in one dispense step change fluids or use another dispenser and dispense to even rows in another dispense step In addition you can combine a selection of colu...

Page 162: ...o rows 1 5 9 13 17 21 and 25 The next section of rows dispensed to includes 2 6 10 14 18 22 26 and 30 And so on creating 4 sections of rows Dispense Pattern with Both Syringes When Both Syringe dispensers are selected to fill a plate the manifolds move in this pattern MultiFlo FX Multi Mode Dispenser ...

Page 163: ...re dispense or tip prime to correct for evaporation and other minor fluid loss to normalize the tips Overfill washing which has proven effective for ELISA assays is a popular feature fluid is aspirated simultaneously to dispensing to prevent spillage by positioning the aspirate tubes at the tops of the wells About the hardware The wash manifolds are identified by the plate types they support e g 9...

Page 164: ...ing fluid removal from the well This option offered as part of the dispense step is called Delay Start of Vacuum Adjust the Aspirate Travel Rate and Aspirate Height when defining the aspirate step select one of the specially designed travel rates that minimize turbulence in the wells Increase the aspirate height to leave more residual fluid in the wells to protect the cell layer Also consider usin...

Page 165: ...side of the wells to reduce turbulence and change the aspirate height to increase the amount of residual fluid left in the well to protect the cell layer See Cell Wash Strategies below Be sure to check the parameters for Final Aspiration Cell Wash Strategies To give you a starting point for optimizing your own assays here are some recommendations for improving your cell wash assays Repositioning t...

Page 166: ...iration or Vacuum On Volume Control Touch Screen Edit the Wash or Dispense Step Advanced Options During regular plate washing aspiration and dispensing occurs simultaneously This allows overflow dispensing because the fluid is aspirated before overflowing the plate Use this control to begin aspiration only after the specified volume is dispensed For cell wash assays specify at least 10 µL well For...

Page 167: ...ersons with pacemakers implants should avoid direct contact Keep all magnetic media watches and sensitive electronic devices away from the magnets Credit cards tapes and disks can be erased in the presence of a magnetic field Bodily harm pinching of hands and skin can result if magnets are not handled correctly Maintain distance between two or more magnets Take advantage of the sample magnetic bea...

Page 168: ...gnetic bead assays Refer to the BioStack Operator s Manual for detailed instructions of the alignment procedure To help get you started 1 Place the magnet in the carrier and a microplate on top of it 2 Launch the BioStack Alignment Utility LHC Touch screen Tools BioStack Utilities Alignment Utility Instrument Next BioStack 3 HOME the BioStack and Begin Realignment 4 Lower the claw until a 0 050 1 ...

Page 169: ...t 6CW for the aspirate travel rate The CW travel rates are designed to minimize disruption to cell layers on the bottom of the well The same principle applies to magnetic bead assays l Adjust the Aspirate height increase the aspirate height setting Z axis which will increase the residual fluid in the wells but also preserve the beads Good results were obtained when keeping the aspirate height arou...

Page 170: ...rate Settings Adjust the Y Position to align the aspirate tubes near the well walls if available Increase the Aspirate Height Z axis leaving more residual volume in the wells 384 well Flat Magnet PN 7103017 The magnetic force approx 4300 Gauss is distributed in a vertical pattern column wise across the plate Adjust the X Position to align the aspirate tubes away from the center of the well near th...

Page 171: ... beads to the bottoms and edges of the wells Every well is circled by 4 magnets Aspirate from the center of the well the default position when using this magnet How to determine the Magnet Adapter Height Offset Use calipers for the best results or another measuring device 1 Put a magnet in the adapter and the type of plate you will be using in place on top of it 2 Measure the distance from the bot...

Page 172: ...nd Quick Wash without adjustments when the magnet is used It applies the specified height value as an offset to all relevant steps Custom or non standard microplates and special adapters for labware may benefit from this setting too If a vessel s height has been its only limitation to processing with the MultiFlo FX i e the vessel s geometry permits the manifold tips to successfully address its we...

Page 173: ...he Syringe Dispenser or Strip Washer to dispense magnetic beads you must remove the magnets that normally hold the dispense manifolds on the instrument Two thumbscrews are shipped with the manifolds to replace the magnets 1 Use the 1 16th hex wrench PN 48713 to loosen the set screw on top of the manifold 2 Take advantage of the magnet s attraction to the hex wrench to remove it from the manifold 3...

Page 174: ...g is not available and RAD 8 to 1 bulk dispensing is not recommended when using the Quick menu you must create a protocol for these features Dispense Cassettes n Important It is imperative that the cassette type setting onboard the instrument match the installed cassette l Install the Dispense Cassette on page 56 l Change the cassette type setting on page 76 l Dispense Cassette Diagram on page 54 ...

Page 175: ...tension tip geometry etc The variation between volumes of fractional aliquots is significantly higher than between dispenses done with full aliquots Thus the best possible performance and reproducibility from dispense to dispense is done in full aliquot increments That is specifying a 1 µL dispense volume when using the 1 µL cassette 5 µL dispense volume when using the 5 µL cassette and 10 µL disp...

Page 176: ...ispense cassette Important information about dispense cassettes When not in use BioTek recommends releasing the tension on the cassette This practice extends its life preserving the tubing s integrity To release the tension 1 Open the Pump Cover 2 Pull out the Cassette Rest 3 Release the spring loaded latch that holds in place the Tube Tensioner attached to the pump s stainless steel plate It will...

Page 177: ... are o 1 µL cassette 1 50 µL or 0 5 µL o 5 µL cassette 5 2500 µL o 10 µL cassette 10 3000 µL Note When using the RAD 8 to 1 bulk dispensing cassette lower the dispense height to ensure fluid hits the wells i e select Advance Options and change the Z axis Select the Flow Rate Cassette Type 1 μL 5 μL 10 μL μL sec tube Low 50 120 140 Medium 60 140 160 High 64 160 180 RAD technology Single channel cas...

Page 178: ... Peri pump primes into the priming trough immediately before dispensing to the plate Pre dispense is recommended for most applications It normalizes the tips to correct for evaporation for example to ensure precise fluid distribution See Protocol Parameters Tables on page 103 for details about the remaining parameters RAD Tip Tracking When using the 8 to 1 bulk dispensing cassette you can further ...

Page 179: ...e will collide with both the RAD priming cup and the Syringe dispenser insert if they are left in the priming trough RAD Well Map RAD technology s single channel dispenser is ideal for partially filling a plate It lets you pick the individual wells to dispense to hit picking Green wells are dispensed to white wells are skipped Quick Dispense Protocol Dispense Step Press Well Map Create Edit the Pr...

Page 180: ... of a message screen interrupting the operation l Automatically Set the cassette type this option changes the MultiFlo FX s setting without a confirmation This option is for advanced users only n No action is taken when the cassette type setting matches the protocol s required cassette type n RAD technology cassettes add a layer of complexity to this feature Error code 6029 is displayed when a mis...

Page 181: ... the offsets will be provided Contact BioTek TAC for assistance if you have questions about dispense performance at 0 5 µL 8 to 1 Tube RAD Cassette Chute height represents the distance between the bottom of the 8 tube tips to the bottom of the chute tip The default value reflects the chute provided with the bulk dispensing cassette Only change this value if you remove or replace the chute Set the ...

Page 182: ...to accurately deliver buffer reagents and other fluids See Syringe Dispenser Autoclavable vs Non autoclavable on page 167 to learn the difference between these two options Prime the Tubing l How to Prime the Syringe dispenser below l Recommended prime volumes for the Syringe dispensers on page 101 l See also Syringe Dispenser and Strip Washer Maintenance on page 201 How to Prime the Syringe dispen...

Page 183: ... protocols for the syringe dispensers e g S DAY_ RINSE fully prime the system pumping 40 mL through the tubing and pump The approximate dead volume for each Syringe dispenser system is 12 mL Generally three times the dead volume completely primes the system When using precious fluids e g expensive reagents you can change the prime parameters reduce the volume or number of cycles specified in the p...

Page 184: ...h inserts with sufficient fluid to cover the tubes e g 6000 µL And be sure to use the priming trough inserts when the option is enabled Learn more about soaking the dispense tubes Set the Submerge Duration and fill the Submerge checkbox to enable this option n Remove the Peri pump cassette or its priming trough insert when soaking the Syringe dispenser or washer manifolds 5 Optionally adjust the P...

Page 185: ...Plates Use Both Syringe dispensers when processing 1536F plates 2 Enter the Volume to dispense per well The valid values are 5 30 000 µL depending on the plate type 3 Define the Flow Rate The rate at which the fluid is dispensed Options range from 1 5 1 is the slowest 5 is the fastest The valid rate is volume dependent Lower rates are recommended for viscous fluids Rates are volume and plate type ...

Page 186: ...Rate SB LB 1 56 125 2 58 150 3 60 162 4 62 174 5 64 187 The default rate is 3 Low density plate manifolds Like the standard manifolds described above for these plate type specific manifolds the flow rate is defined as µL sec well and the minimum volumes are cumulative i e any rate can be used to dispense Rate 5 s minimum Manifold Flow Minimum Volume Rate 1 Rate 2 Rate 3 Rate 4 Rate 5 6 Well Flow 5...

Page 187: ... the tubing s check valves perform more slowly Delaying the syringe pump sufficiently to allow the specified amount of fluid to pass through the check valves before being pumped into the syringe has been shown to improve dispense accuracy Begin by setting the delay to 500 msec Experiment with different settings to determine the optimal value for your fluid Change the Syringe Dispenser Manifold Cha...

Page 188: ...send this setting to the instrument Syringe Dispenser Settings Syringe Dispenser Assembly After installing the Syringe dispenser unit update the instrument s settings The MultiFlo FX automatically detects which type of pump is installed Select the manifold installed l Non Autoclavable the black plastic casing of the non autoclavable syringes are easy to distinguish from the glass and stainless ste...

Page 189: ...ation Data Calibration data is critical to achieving expected performance and it is unique to each individual syringe unit BioTek prints the values on the bottom of the unit so you have a permanent record of the values When installing the Syringe dispenser enter the values printed on the unit Syringe Dispenser Autoclavable vs Non autoclavable Two types of Syringe dispensers are available autoclava...

Page 190: ...view the Help topic LHC Protocols Explained to learn the distinctions To change the onboard settings LHC Touch screen 1 Select Tools Instrument Utilities 2 Choose the applicable tab and modify settings as desired 3 Click Send after changing a setting to update the instrument 1 Press Instrument Options for washer settings 2 Explore the other tabs for more settings n When using the LHC to change ins...

Page 191: ...urrently installed carrier o STD Standard carrier o MTUBE Mini tube carrier Change the Plate Clearance Setting Touch screen The Plate Clearance setting adds the specified value to the travel height for the selected plate type i e it adds this number to the Plate Height value cited in the Plate Types Table The manifold rises to this height to move from one column to the next and whenever reposition...

Page 192: ...efresh the list of protocols onboard the Instrument by clicking the Settings link 3 Highlight one or more protocols in a display box Hold the Ctrl or Shift key to simultaneously select multiple files 4 Optionally at the top left corner of the screen choose to Disable Editing of transferred protocols to lock the protocols from editing or deleting when they are onboard the instrument 5 Click the app...

Page 193: ...them touch or transfer all protocols simultaneously Protocols transferred to the MultiFlo FX can be run from the Home screen and if not locked modified from the Protocol Definition screen Locked files you can unlock a protocol by replacing it using the Protocol Transfer control Upload the protocol to a USB stick make sure Lock files is disabled and re transfer the protocol It will overwrite the ex...

Page 194: ... or USB to copy files from it to the MultiFlo FX 3 Optionally apply the Lock files option to prevent users from changing the protocols 4 Select files individually by highlighting them touch or transfer all files simultaneously Files transferred to the MultiFlo FX can be attached to a protocol and if not locked modified from the Plate Lid Definition screen MultiFlo FX Multi Mode Dispenser ...

Page 195: ...190 Clean the Bottles 190 Clean the Plate Carrier 191 Clean the exterior surfaces 191 Peri pump Dispenser Maintenance 192 Flush the Dispense Cassette 192 Unclog the Dispense Tips 194 RAD Single tube Cassette Unclog the Tip 196 RAD 8 to 1 Cassette Unclog the Tips 197 Record the Number of Plates Processed 199 RAD 8 to 1 Cassette Replace Chute 200 Syringe Dispenser and Strip Washer Maintenance 201 Cl...

Page 196: ... must be based on the risk and performance factors of your assays Develop a maintenance schedule for your MultiFlo FX based on the characteristics of the fluids used and the activity level Here are some guidelines for each component Washer o If the washer will be idle for several hours or days at a time soak the tubes to keep them in a wetted state Enable AutoPrime if the washer is idle for more t...

Page 197: ...Clean the Dispense Aspirate Tubes on page 202 Recommended Maintenance Schedule The schedule recommends preventive maintenance tasks the frequency with which each task should be performed and the predefined onboard Maintenance program that should be run if applicable See Recommended Maintenance Schedule on the next page n It is important to note that the risk and performance factors associated with...

Page 198: ...assette ü ü Record number of plates processed with cassette ü Syringe Dispenser Run S DAY_RIN SE_A B ü ü Run AutoPrime ü Components Remove protein residuals and fungi growth if necessary ü ü ü Check empty waste bottles ü ü Clean bottles ü ü Clean plate carrier system ü ü Clean washer manifold ü ü Clean aspirate and dispense tubes ü ü Clean exterior surfaces ü Clean Syringe dispenser manifold ü ü C...

Page 199: ...te external surfaces ü ü Run W DECONTAMINATE ü ü Run S DECONTAMINATE ü ü Prepare for Storage or Shipment Run W LONG_SHUTDOWN ü Run S LONG_SHUTDOWN ü Replace Repair Components Recalibrate Peri pump cassette As Needed Replace Peri pump dispense tips chute As Needed Replace Peri pump cassette tubing As Needed Replace Syringe dispenser check valves As Needed BioTek Instruments Inc ...

Page 200: ... dispense manifolds require more diligence to keep them clog free Run these protocols and enable AutoPrime to satisfy the daily maintenance requirements l W DAY_RINSE l S DAY_RINSE_A B l P UL_CASS_RINSE represents the cassette type P2 protocols are for the secondary Peri pump Make sure the supply bottles contain sufficient rinse solution and that the waste bottles are empty before running the prot...

Page 201: ...t of your daily maintenance routine About AutoPrime AutoPrime automatically primes the tubing whenever the instrument is idle for a specified time Keeping the tubes wet prevents clogging and mitigates fluid evaporation at the tips AutoPrime s submerge feature lets you soak the tubes for extended periods which is an effective maintenance option The downtime interval is defined for all components bu...

Page 202: ...LHC Touch screen 1 Select Tools Instrument Utilities AutoPrime tab 2 Specify the idle time interval that will trigger an AutoPrime up to 24 hours 3 Enable and define the parameters for each device Remember to set a Submerge Duration to employ this option 4 Click Send to transfer the settings to the instrument 1 Select Instrument at the Home screen and select AutoPrime 2 Specify the idle time inter...

Page 203: ...d it will be moved into the submerge position when the Syringe tips are submerged Either remove the cassette altogether when soaking the Syringe tips or remove the prime trough inserts for the Peri pump to prevent cross contamination of fluids or unintended wicking of fluid into the cassette n Keep in mind that any interaction with the MultiFlo FX will reset the interval clock And AutoPrime only r...

Page 204: ...AutoPrime Interval enter the downtime interval that will trigger an AutoPrime for all devices 3 Fill the checkbox for the dispenser and then press its link 4 Set the Volume and Rate The default settings are 400 µL at rate 3 Note The Volume dispensed is actually double the input value The Syringe dispenser always performs two prime cycles during AutoPrime doubling the volume For example when 400 µL...

Page 205: ...washer and press its link 4 Set the Volume and Rate The default values are 400 µL at rate 3 x2 Two dispense cycles are performed 5 You can Submerge the manifold tubes in the fluid after the prime if desired Set Duration in Hours and Minutes After the dispense tubes have been primed the manifold moves down into the priming trough that is filled with the dispensed solution The tubes are allowed to s...

Page 206: ...move either the cassette or the Peri pump priming trough from the instrument before performing this procedure 5 Run a System Self Test on page 218 to empty the washer s priming trough when restarting the instrument n Note the washer aspirates fluid during the prime step but it does not fully evacuate the priming trough insert 32 tube Syringe Dispenser When using the 32 tube dispense manifold espec...

Page 207: ...trough inserts to submerge the tips n Note the aspirate manifold does not completely evacuate the priming trough insert n Overnight Maintenance You can turn off the instrument after the tips are submerged to leave them soaking in the priming trough for an extended duration 3 Click Run Save the program with a memorable name for future use AutoPrime Enable AutoPrime to ensure the tubing is soaked du...

Page 208: ...you define the prime routine Two priming trough inserts are supplied one for each manifold The troughs can be used to preserve precious fluids and to soak the manifold tubes n Note the washer aspirates fluid during the prime step but it does not fully evacuate the priming trough insert n Be sure to use the prime trough inserts to submerge the tips in fluid Create a Prime protocol As an alternative...

Page 209: ...rime only runs when the main menu quick menu or run completion message is displayed n When AutoPrime is running you can press STOP on the keypad to stop it It will run again the next time the downtime interval occurs Removing Protein Residuals and Fungi Growth Important Solutions containing proteins such as bovine serum albumin BSA will compromise the MultiFlo FX s performance over time unless a s...

Page 210: ...AY_RINSE protocol three times with deionized water if you plan to use the device immediately n N Normal solution which contains 1 gram equivalent weight gEW of solute per liter of solution The gram equivalent weight is equal to the molecular weight expressed as grams divided by the valency of the solute Alternatively use an Enzyme Active Detergent 1 Mix an enzyme active detergent according to the ...

Page 211: ...these components n Water Daily maintenance is the key to keeping the liquid handler performing to specifications In the maintenance procedures provided in this manual the requirement to use distilled dH2O or deionized DI water can be met by numerous water purification methods including MilliQ A minimum water purity of 2mOhm is expected Important l Do not immerse the instrument spray it with liquid...

Page 212: ...s l Autoclaving the cassette typically increases the tubes capacity Expect the cassette to dispense more fluid than expected immediately after sterilizing or disinfecting the tubing When the cassette is completely dry dispense volumes return to normal l Autoclaving the dispense cassette does not diminish its expected life Clean the Bottles l Clean and rinse the supply bottles with deionized water ...

Page 213: ... isopropyl alcohol or ethanol Clean the priming trough too 4 If detergent was used wipe the components with a cloth moistened with water Use a clean dry cloth to dry the components 5 Reinstall the carrier l Place it on the transport rail so the slot on its base fits into the Y axis Carrier Arm l Clip the bottom bracket onto the transport rail align the little bracket on the left rail to hold the c...

Page 214: ...mber of plates processed with a cassette This is necessary to determine when the cassette has reached its expected lifetime and is due for replacement or recalibration Replacement Tubing Kits a refurbishment service and new cassettes are available from BioTek Instruments Monthly maintenance requires overall cleaning of the dispenser and its accessories and verifying performance to determine if the...

Page 215: ...rst prime the tubing with a buffered saline solution to remove any traces of water in the tubing then prime with the dispense fluid 2 Prime the tubing until fluid flows into the priming trough and all visible air bubbles have been removed At the end of the day Purge the tubing to reclaim the dispense fluid then Prime the tubing to flush it clean 1 Purge the cassette until the tubing appears empty ...

Page 216: ...ek ships a 10 cc plastic syringe with special tubing and fitting for use unclogging tips Installation instructions recommend storing it in the pouch on the back of the instrument The remedy involves removing the dispense tip and flushing it with water Depending on the type of clog soaking the tip holder in hot water with mild detergent is recommended This task may be easier if you use the cassette...

Page 217: ...ble the cassette l Push the straight end of the tip into the bottom of the tube the tapered end of the dispense tip is exposed l Reinsert the tube into the Tip Holder Seat the flared edges of the tip into the molded slots l Replace the Tip Holder cover with its two screws The etched BioTek label identifies the top of the cover except for 1536 cassettes steel cover plate RAD Technology Cassettes Ga...

Page 218: ...nstructions to flush the tip using the 10 cc syringe 4 When finished reassemble the tip holder l Put the straight end of the tip into the bottom of the tube leaving the tapered end exposed l 1 µL tubing put the little block in the space provided for it l All tubing slide the flared top of the tip into its molded slot Only the end of the tip protrudes from the holder l Replace the bottom plate it o...

Page 219: ... FX to remove the three screws on the 8 to 1 cassette s cover plate 2 Remove the plate and set it aside 3 Remove the two screws that hold the chute on the cassette 4 Remove the chute and set it aside This frees the tips and tubing from the cassette 5 Pull the tubing off the tips and follow the instructions to use the 10 cc syringe to unclog tips See Unclog the Dispense Tips on page 194 6 When fini...

Page 220: ...post Try to make the tubes flush with the tip head Use a little isopropyl alcohol on the tubing to help it slide onto the post Note Components fit back together in only one way l Attach the tip head with tubing to the cassette head by aligning the pins on the tip head with their sockets on the cassette l Reattach the chute aligning it on the same pins and securing it with its two screws Tighten ea...

Page 221: ...Processed To determine when a tubing cassette has reached the end of its expected lifetime make a habit of counting and recording the approximate number of plates and volume dispensed per cassette Create a form similar to the example table below or estimate your usage of the cassette and project a date for replacement or recalibration Cassette Expected Lifetime Cassette Types Cassette Life Total V...

Page 222: ...Vol Total Cassette Vol 10 10 08 26 384 5 µL 49920 µL 50 mL 10 11 08 33 96 10 µL 31680 µL 82 mL RAD 8 to 1 Cassette Replace Chute As needed you can replace the chute on the 8 tube RAD bulk dispensing cassette A spare chute is included in the cassette package Use the Philips screwdriver shipped with the MultiFlo FX to remove the chute and replace it with another To reattach the chute align it on the...

Page 223: ...articles behind like other materials Clean the Bottles and Tubes l Clean the dispense and rinse bottles and supply tubes with deionized water before the first use before each refill and if they have been idle for any length of time l Accumulated algae fungi or mold may require decontamination Syringe Dispenser Maintenance Tasks l Autoclave the Syringe Head on page 204 l Clean the Dispense Tubes on...

Page 224: ...spense Tubes below if necessary Clean the Dispense Aspirate Tubes n Do NOT autoclave the dual 8 tube and 32 tube dispense manifolds n Do NOT autoclave the washer dispense or aspirate manifolds n Note The autoclavable 16 tube and non autoclavable 8 tube manifolds have removable dispense tubes We do not recommend routinely removing these tubes In the case of a particularly difficult problem with any...

Page 225: ...and 16 tube manifold stylus PN 2872304 o 32 tube manifold 10 cc syringe and tubing Let water flow through the open channel while you probe or flush each tube forcing any particles to be washed away 4 Rinse the manifold with deionized or distilled water Check to see if water comes out of all of the dispense tubes except when working with the 32 tube SB manifold Reinsert the plugs into the ends of t...

Page 226: ...n valve indicating flow direction 3 Flush with hot water 4 Replace the valve and the tubing Replace Syringe Dispenser Check Valve on page 211 if necessary Autoclave the Syringe Head Certain models of the Syringe dispenser are not autoclavable Be sure your dispenser is described as autoclavable before proceeding See Syringe Dispenser Autoclavable vs Non autoclavable on page 167 1 Use the supplied 3...

Page 227: ...6 With the flat side of the shaft facing up slide the syringe piston shaft into the piston holder until it stops 7 Use the 3 32 2 39 mm hex wrench to tighten the setscrew 8 Push the syringe head over the piston until it is flush with the unit and use the hex wrench to attach the two mounting screws Decontamination Any laboratory instrument that has been used for research or clinical analysis is co...

Page 228: ...leaning solution to run into the interior of the instrument If this happens contact the BioTek TAC Do not soak the keypad n Wear prophylactic gloves when handling contaminated instruments Gloved hands should be considered contaminated at all times keep gloved hands away from eyes mouth nose and ears Eating and drinking while decontaminating instruments is not advised n Mucous membranes are conside...

Page 229: ...rd Empty the waste bottle 2 Unload the Peri pump cassette and the prime trough inserts and remove the Syringe dispenser manifold and tubing if applicable 3 Autoclave the cassette and other autoclavable components 4 Prepare an aqueous solution of 0 5 sodium hypochlorite NaClO or bleach As an alternative 70 isopropyl alcohol or 70 ethanol may be used if the effects of bleach are a concern n Isopropy...

Page 230: ...nd dispenser B When storing or shipping the instrument the LONG_SHUTDOWN procedure page 208 primes and soaks the instrument and ends by pushing air through the system The parameters can be edited for optimum cleaning For example consider using ethanol instead of air to complete the decontamination process n Two supply bottles are required for this procedure one for disinfectant and one for rinse 1...

Page 231: ...n this case prepare a 1 20 dilution Household bleach is typically 5 NaClO in this case prepare a 1 10 dilution 1 Turn the instrument off and disconnect the power cord 2 Unload the Peri pump cassette and the prime trough inserts if applicable Clean and store them separately 3 Empty the waste bottle 4 Prepare an aqueous solution of 0 5 sodium hypochlorite NaClO or bleach 5 Fill one supply bottle wit...

Page 232: ...me components of the MultiFlo FX must be replaced periodically to maintain specified performance levels Peri pump Components l Replace Peri pump Dispense Cassette Tubing below l Recalibrate the Peri pump Dispense Cassette on the facing page Syringe Dispenser and Strip Washer Components l Clean or Replace the Check Valves on page 204 l Calibrate the Backlash for Syringe Dispenser on page 212 Replac...

Page 233: ... on the MultiFlo FX Operator s Manual USB stick titled Calibration Jig Instructions 7171009 current Rev pdf Important After calibrating the cassette BioTek recommends performing the Peri Pump Dispense Precision and Accuracy Tests defined in the operator s manual and IQOQPQ procedure to verify the cassette meets specifications Replace Syringe Dispenser Check Valve You can order replacement check va...

Page 234: ...lity of 0 001 g resolution l Supply bottle with deionized water Setup While calibrating and testing try to maintain a steady liquid level in the supply bottle keeping it half full Start with more fluid to allow for priming Connect the supply bottle to the Syringe under test Make sure the supply bottle is at the same level as the dispenser Create a Dispense Protocol Create a protocol that dispenses...

Page 235: ...klash setting use 0033 for autoclavable units use 0031 for non autoclavable units l Subtract the Actual Weight from the Expected Weight and divide by the number of columns dispensed to l Divide the result by 0 0033 and round to the nearest whole number Example 7 680 7 550 0 130 0 130 24 0 00542 0 00542 0 0033 1 64 rounded to 2 The backlash setting needs to be adjusted by 2 7 Adjust the backlash as...

Page 236: ...214 Chapter 4 Maintenance MultiFlo FX Multi Mode Dispenser ...

Page 237: ...rview 216 Qualification Schedule 217 System Self Test Verify Information 218 Liquid Testing the MultiFlo FX Multi Mode Dispenser 219 Peri pump Dispense Precision and Accuracy Tests 221 Perform the Peri Pump Precision and Accuracy Tests 224 Syringe Dispenser Liquid Tests 235 Perform the Syringe Dispense Precision Accuracy Test 236 Washer Qualification Liquid Tests 250 ...

Page 238: ...sher Qualification Tests l Dispense Precision and Accuracy Test on page 253 l Evacuation Efficiency Test on page 254 Peri pump and Syringe Dispensers l Dispense Precision is a measure of the variability of volumes dispensed from tube to tube across the manifold or tip holder The optical density of the solution in a well is proportional to the total volume of the solution in the well When the Coeff...

Page 239: ...tion Qualification Operational Qualification and Performance Qualification IQ OQ PQ and preventative maintenance PM Extensive Checklists and Logbooks are included for recording results Contact your local dealer for more information Tests IQ OQ PQ Initially Annually Monthly Unpacking Installation and Setup ü System Self Test and Checksum Test ü ü ü Shake Test ü Washer Evacuation Efficiency Test ü ü...

Page 240: ...s an error code displays If this happens find the error code in the MultiFlo FX Operator s Manual to determine its cause If the problem is something you can fix turn off the instrument fix the problem and then turn the instrument back on and re run the test If the problem is not something you can fix contact BioTek s Technical Assistance Center Record Onboard Software Record the software part numb...

Page 241: ... Dispense Precision and Accuracy Test For each Peri pump installed perform two tests one at the volume that matches the cassette type and another that best represents the cassette type and dispense volume most common to your applications l RAD Technology Cassettes For the single tube cassettes perform the two tests as recommended for the standard cassettes except limit the dispense to only one row...

Page 242: ...tests using the recommended solutions Plate Reading l If you are using one of BioTek s keypad based readers such as the ELx800 or ELx808 ensure that the reader is not running in Rapid mode To check the setting select UTIL à READ and cycle through the options until READ IN RAPID MODE appears Set it to NO l The absorbance of blue dye solutions should be measured at 630 450 or 405 nm The BioTek blue ...

Page 243: ...lume is 80 µL well n Note For IQ OQ PQ purposes you can add 1 0 additional tolerance to the Precision CV to accommodate various test solutions off peak wavelengths reader errors and pipette errors Peri pump Precision and Accuracy Testing Methodology Tare an empty plate on a balance Use the Peri pump to dispense a quantity of fluid with a known dye concentration to the wells Weigh the plate to obta...

Page 244: ...L cassettes when dispensing 10 aliquots 2 1 2 turns of pump and the 10 µL cassettes when dispensing a single aliquot 1 4 turn of the pump into each well of the plate It dispenses 10 µL into each well using the 10 µL Solution and requires an additional 100 µL of deionized or distilled water to raise the fluid level for optimal reading l 50 µL Test Confirms the performance of the 5 µL cassettes when...

Page 245: ...ng at 630 and 450 or 405 nm l BioTek blue dye solution PN 7773001 or equivalent to create the Peri pump Dispense Precision and Accuracy Test Solutions below n See Important Recommendations for All Liquid Tests on page 220 Peri pump Dispenser Precision and Accuracy Test Solutions Unique concentrations of the test fluid are described here each one corresponds to a specific dispense volume Prepare th...

Page 246: ...d them on the operator s manual USB stick in the Qualification chapter PDF Procedure 1 Install the cassette to be tested 2 Turn on the MultiFlo FX and make sure the cassette type setting is correct 3 Turn on the balance 4 Fill a beaker or other vessel with the test solution 5 Define a Protocol and save it for reuse Set the parameters based on the desired test volume n A predefined protocol for the...

Page 247: ...ll Map to dispense to one row only Then perform the remaining steps in this procedure on that row only Remember to calculate the results based on 12 wells rather than 96 5 Place a clean new microplate on the balance and tare the balance 6 Put the Tube Organizer into the test fluid vessel and Prime the tubing until any large air bubbles are removed 7 Run the dispense protocol 8 Place the plate on t...

Page 248: ...not clogged follow instructions to Unclog the Dispense Tips on page 194 If that doesn t work recalibrate the cassette and repeat the test s If your tests continue to fail contact BioTek s Technical Assistance Center TAC Documenting Test Results Dispense Precision Accuracy Test Worksheets are provided below We recommend you make copies of the appropriate pages and use them to record your calculatio...

Page 249: ...se Accuracy Test Total Dispense Actual Weight grams Expected Weight mL well x number of wells dispensed grams Accuracy Error Actual Weight Expected Weight Expected Weight x 100 Accuracy Error must be 10 0 c Pass c Fail Visual verification that no well varies considerably from the others c Pass c Fail Cassette Serial Number Tests Performed By Date Reviewed Approved By Date MultiFlo FX Qualification...

Page 250: ...nse Accuracy Test Total Dispense Actual Weight grams Expected Weight mL well x number of wells dispensed grams Accuracy Error Actual Weight Expected Weight Expected Weight x 100 Accuracy Error must be 5 0 c Pass c Fail Visual verification that no well varies considerably from the others c Pass c Fail Cassette Serial Num ber Tests Performed By Date Reviewed Approved By Date MultiFlo FX Qualificatio...

Page 251: ...Test Total Dispense Actual Weight grams Expected Weight mL well x number of wells dispensed grams Accuracy Error Actual Weight Expected Weight Expected Weight x 100 Accuracy Error must be 5 0 c Pass c Fail Visual verification that no well varies considerably from the others c Pass c Fail Cassette Serial Num ber Tests Performed By Date Reviewed Approved By Date MultiFlo FX Qualification Procedures ...

Page 252: ...y Test Total Dispense Actual Weight grams Expected Weight mL well x number of wells dispensed grams Accuracy Error Actual Weight Expected Weight Expected Weight x 100 Accuracy Error must be 4 0 c Pass c Fail Visual verification that no well varies considerably from the others c Pass c Fail Cassette Serial Num ber Tests Performed By Date Reviewed Approved By Date MultiFlo FX Qualification Procedure...

Page 253: ...nse Accuracy Test Total Dispense Actual Weight grams Expected Weight mL well x number of wells dispensed grams Accuracy Error Actual Weight Expected Weight Expected Weight x 100 Accuracy Error must be 2 0 c Pass c Fail Visual verification that no well varies considerably from the others c Pass c Fail Cassette Serial Num ber Tests Performed By Date Reviewed Approved By Date MultiFlo FX Qualificatio...

Page 254: ...nse Accuracy Test Total Dispense Actual Weight grams Expected Weight mL well x number of wells dispensed grams Accuracy Error Actual Weight Expected Weight Expected Weight x 100 Accuracy Error must be 4 0 c Pass c Fail Visual verification that no well varies considerably from the others c Pass c Fail Cassette Serial Num ber Tests Performed By Date Reviewed Approved By Date MultiFlo FX Qualificatio...

Page 255: ...racy Test Total Dispense Actual Weight grams Expected Weight mL well x number of wells dispensed grams Accuracy Error Actual Weight Expected Weight Expected Weight x 100 Accuracy Error must be 2 0 c Pass c Fail Visual verification that no well varies considerably from the others c Pass c Fail Cassette Serial Num ber Tests Performed By Date Reviewed Approved By Date MultiFlo FX Qualification Proced...

Page 256: ...Accuracy Test Total Dispense Actual Weight grams Expected Weight mL well x number of wells dispensed grams Accuracy Error Actual Weight Expected Weight Expected Weight x 100 Accuracy Error must be 4 0 c Pass c Fail Visual verification that no well varies considerably from the others c Pass c Fail Cassette Serial Num ber Tests Performed By Date Reviewed Approved By Date MultiFlo FX Qualification Pr...

Page 257: ...ell 5 20 µL well 2 96 well plate 3 CV 160 µL well 1 25 160 µL well 1 8 16 tube manifold 6 CV 40 µL well 5 40 µL well 3 1536 well plate 12 CV 6 µL well 5 6 µL well 32 tube manifold Low Density Plate Specific Manifolds Dispense Precision Refer to the values below for dispense precision when measured in full plates with the specified volume of deionized water with 0 1 Tween 20 and FD C 1 blue dye sol...

Page 258: ...and 405 or 450 nm l The test solutions Syringe Dispenser Test Solutions below n See also Important Recommendations for All Liquid Tests on page 220 Syringe Dispenser Test Solutions 20 µL Solution o Mix 10 mL of BioTek s blue dye solution with 100 mL of deionized or distilled water to create a dilution of the concentrate Mix 160 mL of deionized or distilled water with 20 mL of the diluted concentra...

Page 259: ...e using the applicable plate type and dye solutions Plate Type Specific Manifolds Perform one test for each syringe using the applicable plate type and the 80 µL solution Standard Syringe Manifolds Low Density Plate Type Specific Manifolds 384 well for the 16 tube manifolds unless only 96 well plates are used in your lab 1536 well for the 32 tube manifolds and 96 well for the dual 8 tube manifold ...

Page 260: ...6 Well 12 Well 24 Well 48 Well Volume µL well 5560 2240 1120 560 Flow Rate 5 5 5 5 Z axis Dispense Height 465 460 452 460 Pre dispense 2 cycles 320 220 160 320 2 Save the protocol 4 Place a clean empty microplate on the balance and tare the balance 5 Place the microplate on the carrier and run the protocol created in step 3 or predefined 6 Place the plate on the balance and record the Total Dispen...

Page 261: ...d Weight Expected Weight x 100 Subtract the expected dispense weight see below from the Actual Total Dispense Weight from step 5 and divide the result by the expected weight Multiply the result by 100 The Expected Dispense Weight is the volume dispensed per well in mL multiplied by the number of wells dispensed For example if 40 μL is dispensed to 96 wells the expected weight is 0 040 x 96 3 84 gr...

Page 262: ...ith the stylus re prime the manifold and repeat the test s If your tests continue to fail contact BioTek s Technical Assistance Center Documenting Test Results Dispense Precision Accuracy Test Worksheets are provided on the MultiFlo FX Operator s Manual USB flash drive in the Qualification chapter PDF We recommend you make copies of the appropriate pages and use them to record your calculations an...

Page 263: ...L Dispense Accuracy Test Total Dispense Actual Weight grams Expected Weight mL well x number of wells dispensed grams Accuracy Error Actual Weight Expected Weight Expected Weight x 100 Accuracy Error must be 5 0 c Pass c Fail Visual verification that no well varies considerably from the others c Pass c Fail Serial Number Tests Performed By Date Reviewed Approved By Date MultiFlo FX Qualification P...

Page 264: ...µL Dispense Accuracy Test Total Dispense Actual Weight grams Expected Weight mL well x number of wells dispensed grams Accuracy Error Actual Weight Expected Weight Expected Weight x 100 Accuracy Error must be 1 25 c Pass c Fail Visual verification that no well varies considerably from the others c Pass c Fail Serial Number Tests Performed By Date Reviewed Approved By Date MultiFlo FX Qualification...

Page 265: ...µL Dispense Accuracy Test Total Dispense Actual Weight grams Expected Weight mL well x number of wells dispensed grams Accuracy Error Actual Weight Expected Weight Expected Weight x 100 Accuracy Error must be 5 0 c Pass c Fail Visual verification that no well varies considerably from the others c Pass c Fail Serial Number Tests Performed By Date Reviewed Approved By Date MultiFlo FX Qualification ...

Page 266: ...µL Dispense Accuracy Test Total Dispense Actual Weight grams Expected Weight mL well x number of wells dispensed grams Accuracy Error Actual Weight Expected Weight Expected Weight x 100 Accuracy Error must be 1 25 c Pass c Fail Visual verification that no well varies considerably from the others c Pass c Fail Serial Number Tests Performed By Date Reviewed Approved By Date MultiFlo FX Qualification...

Page 267: ...µL Dispense Accuracy Test Total Dispense Actual Weight grams Expected Weight mL well x number of wells dispensed grams Accuracy Error Actual Weight Expected Weight Expected Weight x 100 Accuracy Error must be 5 0 c Pass c Fail Visual verification that no well varies considerably from the others c Pass c Fail Serial Number Tests Performed By Date Reviewed Approved By Date MultiFlo FX Qualification ...

Page 268: ...d Deviation Mean OD x 100 CV 6 0 q Pass q Fail Dispense Accuracy Test Total Dispense Actual Weight grams Expected Weight µL well x number of wells dispensed grams Accuracy Error Actual Weight Expected Weight Expected Weight x 100 Accuracy Error must be 1 0 c Pass c Fail Visual verification that no well varies considerably from the others c Pass c Fail Date Test Performed By MultiFlo FX Qualificati...

Page 269: ...rd Deviation Mean OD x 100 CV 4 0 q Pass q Fail Dispense Accuracy Test Total Dispense Actual Weight grams Expected Weight µL well x number of wells dispensed grams Accuracy Error Actual Weight Expected Weight Expected Weight x 100 Accuracy Error must be 1 0 c Pass c Fail Visual verification that no well varies considerably from the others c Pass c Fail Date Test Performed By MultiFlo FX Qualificat...

Page 270: ...rd Deviation Mean OD x 100 CV 4 0 q Pass q Fail Dispense Accuracy Test Total Dispense Actual Weight grams Expected Weight µL well x number of wells dispensed grams Accuracy Error Actual Weight Expected Weight Expected Weight x 100 Accuracy Error must be 1 0 c Pass c Fail Visual verification that no well varies considerably from the others c Pass c Fail Date Test Performed By MultiFlo FX Qualificat...

Page 271: ...rd Deviation Mean OD x 100 CV 4 0 q Pass q Fail Dispense Accuracy Test Total Dispense Actual Weight grams Expected Weight µL well x number of wells dispensed grams Accuracy Error Actual Weight Expected Weight Expected Weight x 100 Accuracy Error must be 1 0 c Pass c Fail Visual verification that no well varies considerably from the others c Pass c Fail Date Test Performed By MultiFlo FX Qualificat...

Page 272: ...avelengths reader errors and pipette errors Accuracy Error is calculated for the dispense volume specified in the respective test procedure in adherence to the published specifications Evacuation Efficiency Test The Evacuation Efficiency test measures the residual volume mean residual weight per well after aspiration is performed The lower the residual per well the better the evacuation efficiency...

Page 273: ...valent l Precision balance with minimum capacity of 100 g and readability of 0 001 g resolution l Pipettes and graduated beakers l Microplate absorbance reader capable of dual wavelength reading at 630 450 nm l Liquid Test Worksheets at the end of this chapter for recordingdata and results l Deionized or distilled water l Test solutions described below n These volumes are sufficient for performing...

Page 274: ...eionized or distilled water with 20 mL of Solution 2 Create Washer Qualification Protocols 1 Select Maintenance at the Home screen and then Define 2 Press Create 3 Press the Name field and assign a unique name e g 6 Well Dispense Test 4 Plate Type select the appropriate plate type for the test and press OK 5 Add steps with the recommended parameters shown below l Add a W Dispense step for the Disp...

Page 275: ...o perform the Evacuation Efficiency Test n Prerequisite for plate types other than 96 Well Create the protocols needed to test your manifold See Create Washer Qualification Protocols on the previous page One time only Predefined protocols are provided with the instrument qualification package PN 1260521 1 Fill a supply bottle with 2 liters of deionized or distilled water and run the W DAY_RINSE pr...

Page 276: ... greater than 3 0 one or more dispense tubes may need to be cleaned Remove the manifold and use the stylus to clean the dispense tube s giving lower than average absorbance readings When finished re prime the washer and retry the test 9 When finished prime with deionized water to flush out the dye solution Evacuation Efficiency Test n Save the plate When this test is complete use the plate to perf...

Page 277: ...the plate and weigh it immediately because evaporation will affect the results This is the Total Residual Weight in grams 6 Visually inspect the plate and note if any wells appear to have considerably more liquid than others 7 Use the Evacuation Efficiency Test Worksheet to perform data reduction o Divide the Total Residual Weight by number of wells to find the Mean Residual Weight o The Mean Resi...

Page 278: ... the standard Evacuation Efficiency Test fails twice when using a 96 well plate This test will confirm which aspirate tube s may be clogged or if the plate s alignment or position is the problem For a ll other plate types a close examination of the residual volume in each well will identify which aspirate tube s are clogged 1 If you have not already done so repeat steps 2 through 7 of the standard...

Page 279: ...ing the problem o If the problem appears to be related to particular wells clean the associated aspiration tubes remove the manifold and thoroughly clean the tubes with the stylus and repeat the test o If the problem appears to be related to a particular region edge or corner of the plate review the alignment and flatness of the plate on the carrier o For additional suggestions See Troubleshooting...

Page 280: ...258 Chapter 5 Qualification MultiFlo FX Multi Mode Dispenser ...

Page 281: ...n Standard Deviation Mean OD x 100 CV 3 0 q Pass q Fail Dispense Accuracy Test Total Dispense Actual Weight grams Expected Weight mL well x number of wells dispensed grams Accuracy Error Actual Weight Expected Weight Expected Weight x 100 Accuracy Error must be 3 0 c Pass c Fail Visual verification that no well varies considerably from the others c Pass c Fail Date Test Performed By MultiFlo FX Qu...

Page 282: ...otal Residual Weight 96 g Mean Residual Weight 0 002 g q Pass q Fail Evacuation Diagnostics Test check here q if not performed Mean OD for the plate Sum of all wells 96 Residual Factor Mean OD Mean Residual Weight Calculate the Residual Weight for each well well OD Residual Factor Every Residual Weight per well 0 002 g q Yes q No Date Test Performed By MultiFlo FX Qualification Procedures BioTek I...

Page 283: ...Standard Deviation Mean OD x 100 CV 4 0 q Pass q Fail Dispense Accuracy Test Total Dispense Actual Weight grams Expected Weight mL well x number of wells dispensed grams Accuracy Error Actual Weight Expected Weight Expected Weight x 100 Accuracy Error must be 3 0 c Pass c Fail Visual verification that no well varies considerably from the others c Pass c Fail Date Test Performed By MultiFlo FX Qual...

Page 284: ...tion Mean OD x 100 CV 6 0 q Pass q Fail Dispense Accuracy Test Total Dispense Actual Weight grams Expected Weight µL well x number of wells dispensed grams Accuracy Error Actual Weight Expected Weight Expected Weight x 100 Accuracy Error must be 3 0 c Pass c Fail Visual verification that no well varies considerably from the others c Pass c Fail Date Test Performed By MultiFlo FX Qualification Proc...

Page 285: ...ndard Test Total Residual Weight g Verification that wells are consistent in appearance q Pass q Fail Mean Residual Weight Total Residual Weight 6 g Mean Residual Weight 0 6 g q Pass q Fail Date Test Performed By MultiFlo FX Qualification Procedures BioTek Instruments Inc ...

Page 286: ...ation Mean OD x 100 CV 4 0 q Pass q Fail Dispense Accuracy Test Total Dispense Actual Weight grams Expected Weight µL well x number of wells dispensed grams Accuracy Error Actual Weight Expected Weight Expected Weight x 100 Accuracy Error must be 3 0 c Pass c Fail Visual verification that no well varies considerably from the others c Pass c Fail Date Test Performed By MultiFlo FX Qualification Pro...

Page 287: ...dard Test Total Residual Weight g Verification that wells are consistent in appearance q Pass q Fail Mean Residual Weight Total Residual Weight 12 g Mean Residual Weight 0 15 g q Pass q Fail Date Test Performed By MultiFlo FX Qualification Procedures BioTek Instruments Inc ...

Page 288: ...ation Mean OD x 100 CV 4 0 q Pass q Fail Dispense Accuracy Test Total Dispense Actual Weight grams Expected Weight µL well x number of wells dispensed grams Accuracy Error Actual Weight Expected Weight Expected Weight x 100 Accuracy Error must be 3 0 c Pass c Fail Visual verification that no well varies considerably from the others c Pass c Fail Date Test Performed By MultiFlo FX Qualification Pro...

Page 289: ...dard Test Total Residual Weight g Verification that wells are consistent in appearance q Pass q Fail Mean Residual Weight Total Residual Weight 24 g Mean Residual Weight 0 05 g q Pass q Fail Date Test Performed By MultiFlo FX Qualification Procedures BioTek Instruments Inc ...

Page 290: ...ation Mean OD x 100 CV 4 0 q Pass q Fail Dispense Accuracy Test Total Dispense Actual Weight grams Expected Weight µL well x number of wells dispensed grams Accuracy Error Actual Weight Expected Weight Expected Weight x 100 Accuracy Error must be 3 0 c Pass c Fail Visual verification that no well varies considerably from the others c Pass c Fail Date Test Performed By MultiFlo FX Qualification Pro...

Page 291: ...dard Test Total Residual Weight g Verification that wells are consistent in appearance q Pass q Fail Mean Residual Weight Total Residual Weight 48 g Mean Residual Weight 0 025 g q Pass q Fail Date Test Performed By MultiFlo FX Qualification Procedures BioTek Instruments Inc ...

Page 292: ......

Page 293: ...Chapter 6 Troubleshooting This chapter provides guidelines for error recovery and troubleshooting performance problems Troubleshooting 272 ...

Page 294: ...an error message that is displayed on the computer screen or keypad The most common error for new MultiFlo FX users is easily fixed 306 Peri pump Pump Cover is open Close the pump cover and re run protocol To run the Peri pump its pump cover must be closed protecting both the pump and the operator 401 Carrier Y motor failed positional verify If the plate carrier is not installed correctly this ins...

Page 295: ...le Cause What To Do Syringe or manifold position error Syringe or manifold is being obstructed Remove obstruction Motor sensor or electrical problem Turn instrument off wait at least 15 seconds turn it back on and run a Quick Dispense routine e g Prime If the problem persists contact BioTek TAC Syringe piston not seated all the way to the bottom before fastening set screw Reinstall the syringe hea...

Page 296: ...t tube to the top port of the syringe pump Check valve flow direction is incorrect Compare the flow direction of the check valves See Syringe Dispenser Check Valves on page 70 Check valves are stuck closed Clean or Replace the Check Valves on page 204 No fluid Fill bottles with appropriate fluid System not primed Run S DAY_RINSE once or twice for one or both syringes Highly viscous fluid Apply a P...

Page 297: ...iston Reinstall the syringe head as described in Autoclave the Syringe Head on page 204 32 tube dispense manifolds are not dispensing accurately Calibrate the Backlash for Syringe Dispenser on page 212 Highly viscous fluid Apply a Pump Delay to improve performance when dispensing highly viscous fluid i e edit the dispense or prime step to set a 500 msec delay Dripping dispense tubes Dispense tubin...

Page 298: ...Carrier Movement Problem Possible Cause What To Do Dispense tubes not entering well correctly Microplate not properly seated or strips not level Reseat microplate carrier or the plate or strips in holder Make sure the carrier is clean Horizontal dispense position does not align the tubes in the wells Change the X axis horizontal Position in the protocol Dispense tube s bent Push the supplied stylu...

Page 299: ... the Dispense Height Uneven dispensing Make sure all cassette components are properly seated in their respective positions Tips are clogged See Unclog the Dispense Tips on page 194 Recalibrate the Peri pump Dispense Cassette on page 211 Replace Peri pump Dispense Cassette Tubing on page 210 Dispenser skipping columns Check define the plate map i e See also Advanced Dispense Options for Peri pump T...

Page 300: ...ke sure the carrier is clean Try a different microplate or strip holder If the problem is unresolved the carrier may have to be realigned Contact BioTek TAC Aspirate tubes position is too wide for a movement Change the horizontal X or Y axis aspirate position in the protocol Aspirate tubes bent Contact BioTek TAC Cell layer disruption Fluid sloshing in wells disturbs cells Slow down the plate carr...

Page 301: ...mize protocols to improve evacuation on page 101 Aspiration tubes not properly positioned horizontally in wells If none of the tubes are bent try adjusting the horizontal aspirate position X Y axis in the protocol Microplate not level in carrier or strips not level in holder Reseat microplate in carrier or strips in holder Make sure the carrier is clean Try a different microplate or strip holder I...

Page 302: ... actual flow direction Check valves are stuck closed Clean or replace the check valves See Clean or Replace the Check Valves on page 204 No wash or rinse fluid Fill bottles with appropriate fluid Ensure bottles are clean and do not contain particles or organic material Unable to dispense fluid System not primed Large air pockets in tubing Run W DAY_RINSE multiple times Insufficient suction clogged...

Page 303: ... it is not turning on contact BioTek TAC If the vacuum pump turns on remove the vacuum tubing from the back of the instrument and put your finger over the port If there is no vacuum contact BioTek TAC Uneven dispensing of fluid wells not filled Clogged dispense tubes on the washer manifold Clean the Washer Manifolds Manifold or tubing not adequately primed Run W DAY_RINSE once or twice Dispense fl...

Page 304: ...ir leaks in the waste tubing and bottle Use a slower Aspiration Travel Rate Enable Continuous Vacuum During Protocol Instrument Washer Uneven not level surface Make sure the washer sits on a perfectly level surface Fluid leaking from underneath the washer module Defective tubing connector or inlet tubing Contact BioTek TAC Leaking valve or syringe seal Contact BioTek TAC Fluid leaking from externa...

Page 305: ...Appendix A Error Codes A listing of potential error codes and possible solutions for resolving them System Error Codes 284 MultiFlo FX Software Error Codes 294 ...

Page 306: ...sing mild detergent and hot water 70 isopropyl alcohol or ethanol Restart the instrument If the error occurs again contact BioTek TAC 211 221 Carrier Y motor didn t find home opto sensor transition Carrier Y motor didn t find autocal jig opto sensor transition Run self test If error reoccurs contact BioTek TAC 212 222 Dispense head motor didn t find home opto sensor transition Dispense head motor ...

Page 307: ...or didn t find autocal jig optical sensor transition Service Only Contact BioTek TAC 300 Carrier X motor interlock safety switch open Service Only Contact BioTek TAC 301 Carrier Y motor interlock safety switch open Service Only Contact BioTek TAC 302 Dispense head motor interlock safety switch open Service Only Contact BioTek TAC 303 Secondary Peri pump motor interlock safety switch open Service O...

Page 308: ...rify Service Only Contact BioTek TAC 406 Peri pump motor failed positional verify Service Only Contact BioTek TAC 407 Strip washer syringe motor failed positional verify Service Only Contact BioTek TAC 408 Aspirate head motor failed positional verify Service Only Contact BioTek TAC 409 RAD Y axis motor failed positional verify Service Only Contact BioTek TAC 600 609 Specified motor currently in us...

Page 309: ... size error not 256 Contact BioTek TAC 1005 Invalid processor signature not 1280 1281 2560 2561 Contact BioTek TAC 1006 Bootcode memory exceeded Contact BioTek TAC 1007 Invalid slave port Contact BioTek TAC 1008 Invalid response from slave Contact BioTek TAC 1009 Invalid processor detected Contact BioTek TAC 1010 Checksum error downloading basecode Contact BioTek TAC 1250 Internal RAM test error o...

Page 310: ... Syringe prime volume 1310 Invalid Syringe manifold 1350 Peri pump PP invalid dispense volume Contact BioTek TAC 1351 PP invalid cassette type Change the cassette type to match the protocol requirement and rerun the protocol 1352 PP invalid pre dispense volume Contact BioTek TAC 1353 Multiple required cassettes Make sure every Peri pump step in the protocol calls for the same cassette type A confl...

Page 311: ... Review the protocol parameters and instrument settings to identify a correction 1407 Invalid step type The protocol may have been created for a different instrument it is not compatible with this instrument 1408 Invalid plate geometry Contact BioTek TAC 1409 Invalid plate carrier type Contact BioTek TAC 1412 Tip clearance error Contact BioTek TAC 1413 AutoPrime in progress Contact BioTek TAC 1414...

Page 312: ...e current hardware 1513 Plate type manifold conflict Change the Plate Type to one supported by the manifold Or check the Instrument Settings and make sure they match the physical hardware Get settings from instrument 1600 160C The onboard storage space allocated for this function has been used up Use the LHC Manage Memory control to reallocate space 160D Not a valid step Contact BioTek TAC 2400 Pa...

Page 313: ...ype in byMsgStructure header field Contact BioTek TAC 8106 Invalid destination in byMsgDestination header field Contact BioTek TAC 8107 Request object received not supported by instrument Contact BioTek TAC 8108 Message Body size exceeds max limit Contact BioTek TAC 8109 Max number of requests currently running and cannot run the latest request Contact BioTek TAC 810A No request running when respo...

Page 314: ...ly level error Service Only Contact BioTek TAC A305 42v Secondary Peri pump motor power supply level error Service Only Contact BioTek TAC A400 Malloc failed Service Only Contact BioTek TAC A500 Multiple tasks attempted to use display simultaneously Service Only Contact BioTek TAC A600 Serial EEPROM access error Service Only Contact BioTek TAC A700 Motor truncation error X axis Carrier X axis moto...

Page 315: ...C A706 Motor truncation error internal primary Peri pump motor steps are being truncated Service Only Contact BioTek TAC A707 Motor truncation error strip washer syringe motor steps are being truncated Service Only Contact BioTek TAC A708 Motor truncation error strip washer aspirate head motor steps are being truncated Service Only Contact BioTek TAC AA30 AA38 Motor profile errors Service Only Con...

Page 316: ...elected for the specified plate type Modify the protocol to select a row 6005 Invalid row selection value must be 0 or 1 Contact BioTek TAC 6006 This instrument can only process 96 well plates The protocol may have been created for another instrument change the plate type or select another protocol 6007 This instrument can only process 1536 well plates 6008 The 8 tube Syringe Manifold can only be ...

Page 317: ...zontal dispense position 6020 Invalid dispense height 6021 Invalid plate clear height 6022 Invalid column selection value must be 0 or 1 6023 Invalid protocol step type 6024 The Definition String contains invalid data 6025 Manifold conflict between protocol requirements and instrument configuration Change the Washer Manifold or change the Instrument Setting 6026 Valve module conflict between proto...

Page 318: ...rted by the washer dispenser Or click the Instrument Settings link and make sure they match the physical hardware Get settings from instrument 6034 The 32 tube Syringe Manifold is required for 1536 well plates 6035 The 16 tube Syringe Manifold is required for 96 and 384 well plates 6036 The 128 tube Washer Manifold is required for 1536 well plates 6037 The 128 tube Washer Manifold can only be used...

Page 319: ...t l Check cables plug in only one communication cable at a time USB or serial l Try running a different protocol If error reoccurs contact BioTek TAC 6050 Invalid message header 6051 Invalid message object 6052 Invalid message body size 6053 Serial message timeout 6054 Port handle error 6055 Read timeout value is invalid 6056 Unauthorized to open the COM port Make sure the COM port setting is corr...

Page 320: ...ected software error that cannot be fixed without BioTek support 6071 Invalid number of syringe prime cycles 6072 Invalid syringe Aspirate Delay value 6073 Invalid X axis offset value 6074 Invalid Y axis offset value Service Only Contact BioTek TAC These codes indicate an unexpected software error that cannot be fixed without BioTek support 6075 Invalid Z axis offset value 6080 Invalid Peri pump p...

Page 321: ...ysically install the Peri pump and or make sure the instrument settings reflect the current state 6160 Strip washer required to perform this step but not installed Make sure the protocol you are trying to run is supported by your instrument s devices 6161 6162 Strip washer manifold and plate type are incompatible Strip washer doesn t support this plate type Make sure the specified plate type match...

Page 322: ...specified plate type matches the installed manifold and the instrument s manifold setting correctly identifies the installed manifold 6171 The installed RAD cassette does not support this plate type Either use a supported plate type correct the instrument s cassette type setting or edit the protocol MultiFlo FX Multi Mode Dispenser ...

Page 323: ...tack errors 272 BioTek s Customer Resource Center 26 Bottle holder 60 Bottom Wash 114 BSA 187 Buffer Switching 4 C Calibration Data 167 212 Carrier setting 169 Cassette Calibration 210 Diagram 54 RAD technology 58 Replacement tubing 13 Shorten tubing 61 Cassette Life 19 22 199 Cassette Types 19 Cell Wash 111 142 aspirate rate 112 Strategy to preserve cells 143 Center holder 54 Change Cassette type...

Page 324: ...uid Leaking 276 282 G General Settings 41 73 H H2O 175 189 Half microliter dispensing 155 Hardware 18 Hardware change detected 96 Highly viscous fluids 162 Hit pick 157 Horizontal aspirate position 278 How it works 153 How To Change cassette setting 76 Change the plate carrier setting 169 Change the plate clearance setting 170 Humidity limits 18 I Incubate 117 118 Incubate a plate at room tem pera...

Page 325: ... diagram 9 Plate Map 157 165 Plate Type 9 supported 18 Plates special handling 134 Plates with lids 126 Port link 120 Power Supply 18 Pre Dispense 113 156 Preserve expensive reagents 61 99 162 Prevent fluid loss during wash 144 Prime Peri pump recommendations 99 Syringe recommendations 101 tips 113 Prime Trough Inserts 59 186 Priming cups 59 Prompt 117 Protein Preventing clogging 187 Protocol disp...

Page 326: ...mended prime volumes 101 Syringe Dispenser Settings 166 Syringe Manifold Change 75 165 Syringe Prime Step 161 Syringe Settings 75 165 System Error Codes 284 T Test solutions 251 Test Solutions Syringe 236 Tests which tests to perform 219 Theory of operation 153 Timer 118 Tip guard 54 Tip holder 54 Tip priming 113 Transfer protocols 170 Troubleshooting 272 Instrument Error Codes 284 LHC Error Codes...

Page 327: ...05 Washer Fixing problems 278 Protocol parameters listing 108 Washer Dispense step 113 Washer Flow Rates 111 Washer Prime step 183 Washer Manifold AutoPrime 183 Water 175 189 Well map 157 BioTek Instruments Inc ...

Page 328: ... only 8 Polycarbonate W Plate carrier P Plate positioning dog peristaltic pump cover latch 9 Polyethylene S W Supply Bottle 10 Polypropylene S W Outlet fitting fittings in bottles inline fittings S W Bottle cap and fittings check valves organic solvent tubing set P Molded dispense tips 11 Polystyrene Assay plates P Peristaltic pump cover Materials listed as 1 16 in Table 1 are cross referenced as ...

Page 329: ...1 chute S W Manifold manifold plugs priming trough insert 15 Silicone P Tubing S W Tubing manifold o rings check valve o rings 16 Ultem polyetherimide P Center Holder Tube Tensioner Tip Holder S W Check valves Note Proper use of vaporized hydrogen peroxide also known as hydrogen peroxide vapor HPV and by the trademarked name VHP in non condensing mode is an approved method of decontaminating the i...

Page 330: ...roxide 10 B A A D A A A A A A A A A C A A Isopropyl Alcohol 70 A A A A A A A A B A A A B A A A Methyl Alcohol 70 A A A A A A A B A A ND A A A A A Methylene Chloride B C A A N D A D D D B D A D A D D Phosphoric Acid 10 D C ND D A A A A A A B A B A C A Propylene Glycol B B ND ND N D ND ND B A A A A C A A ND Sodium Chlorate B C ND ND N D A A A N D A ND A A A C ND Sodium Hydroxide 20 B D A B B A A A A...

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