Biotek Synergy HT Operator'S Manual Download Page 57

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Chapter 3

Getting Started

This chapter describes some of the Synergy™ HT’s key components
and provides an introduction to using Gen5™ or KC4™ to control

the instrument.

Key Components .................................................................. 36

Power Switch, Carrier Eject Button, Microplate Carrier........... 36

Lamp Assembly and Filter Wheel Access.............................. 37

Excitation and Emission Filter Wheels ................................. 38

Installing the Time-Resolved Fluorescence Cartridge ............. 41

Configuring the System for Luminescence Measurements ...... 42

The External Dispense Module ........................................... 43

Gen5™ Software .................................................................. 46

Viewing/Updating the Filter and Wavelengths Tables............. 46

Creating Protocols and Experiments.................................... 47

Controlling the Dispense Module ........................................ 50

KC4™ Software.................................................................... 52

Viewing/Updating the Filter and Wavelengths Tables............. 52

Creating Protocols............................................................ 53

Reading Plates ................................................................ 55

Controlling the Dispense Module ........................................ 56

Recommendations for Achieving Optimum Performance ............. 59

Summary of Contents for Synergy HT

Page 1: ...Operator s Manual Multi Mode Microplate Reader Synergy HT ...

Page 2: ......

Page 3: ...Synergy HT Multi Mode Microplate Reader Operator s Guide November 2008 2008 Part Number 7091000 Revision I BioTek Instruments Inc ...

Page 4: ...ion of BioTek Instruments Inc Trademarks BioTek is a registered trademark and Synergy HT Gen5 KC4 and BioStack are trademarks of BioTek Instruments Inc All other trademarks are the property of their respective holders Restrictions and Liabilities Information in this document is subject to change and does not represent a commitment by BioTek Instruments Inc Changes made to the information in this d...

Page 5: ...ort Service 6 Technical Assistance Center TAC 6 Returning Instruments for Service Repair 6 Contacting BioTek for Applications Support 6 Chapter 2 Installation 7 1 Unpack and Inspect the Reader 8 2 Remove the Shipping Panel 10 3 Remove the Microplate Carrier Shipping Screw 12 4 Install the Fluorescence Lamp Assembly 13 5 Select an Appropriate Location 14 6 Connect the Power Supply 15 7 Unpack and I...

Page 6: ...ls and Experiments 47 Controlling the Dispense Module 50 KC4 Software 52 Viewing Updating the Filter and Wavelengths Tables 52 Creating Protocols 53 Reading Plates 55 Controlling the Dispense Module 56 Recommendations for Achieving Optimum Performance 59 Chapter 4 Instrument Qualification 61 Overview 62 IQ OQ PQ 62 Recommended Qualification Schedule 64 System Test 65 Description 65 Procedure 66 Ab...

Page 7: ...mmended Maintenance Schedule 124 Overview 124 Dispense Module 124 Schedule 126 Warnings Precautions 126 Cleaning Exposed Surfaces 127 Inspecting Cleaning Excitation and Emission Filters 128 Flushing Purging the Fluid Path 129 Running a Dispense Protocol Optional 130 Emptying Cleaning the Tip Priming Trough 132 Cleaning the Priming Plate 132 Cleaning the Internal Components 133 Required Materials 1...

Page 8: ...Lines 158 Rinse the Fluid Lines 159 Clean the Internal Tubing and Injector Heads 159 Clean the Tip Prime Trough and Priming Plate 160 Alternate Procedure for Models with Injectors 161 Appendix B Computer Control 163 Appendix C Error Codes 165 Appendix D Specifications 207 Appendix E Instrument Dimensions for Robotic Interface 215 ...

Page 9: ...et www biotek com Phone 888 451 5171 toll free in the U S 802 655 4740 outside the U S Fax 802 655 7941 E Mail customercare biotek com Service TAC Phone 800 242 4685 toll free in the U S 802 655 4740 outside the U S Fax 802 654 0638 E Mail tac biotek com European Coordination Center Authorized European Representative BioTek Instruments GmbH Kocherwaldstrasse 34 D 74177 Bad Friedrichshall Germany I...

Page 10: ...bance reading speed information p 1 5 Clarified fluorescence specifications p 1 8 and 1 9 Added specifications for injector model p 1 10 Chapter 2 Clarified description of external and internal components of the injector model and updated drawings Moved procedure for replacing the lamp assembly to Ch 6 Chapter 5 Added note regarding the availability of the Installation Operational Performance IQ O...

Page 11: ... Added preventive maintenance procedure for periodic cleaning of the top bottom fluorescence optical probes and the absorbance read channel optical path Appendix A Updated drawing of tip priming trough and priming plate G 8 2005 Updated Warranty information Moved Specifications from Chapter 1 to Appendix D Corrected operating temperature 18 40 C and injector accuracy 1 µl at 5 50 µl to match publi...

Page 12: ...d Added introductory information for new Gen5 users Chapter 4 Instrument Qualification Added instructions for performing the System Test Absorbance Plate Test and Dispense Accuracy Precision Test Chapter 5 Preventive Maintenance Added instructions for creating the optional Dispense protocol in Gen5 I 11 2008 Throughout Changed product description from Multi Detection to Multi Mode Changed Bio Stac...

Page 13: ...section Corrected the formula for Accuracy Error Chapter 5 Preventive Maintenance Removed unnecessary Clean Supply Bottle section Modified the Running a Dispense Protocol procedure to include running the experiment and inspecting the plate Added a missing word to the Cleaning the Optical Probes section Appendix C Error Codes Corrected the range of error codes under Home Sensor Initial Find Errors ...

Page 14: ...w to avoid the problem Caution A Caution indicates potential damage to the instrument and tells you how to avoid the problem Note Bold text is primarily used for emphasis italic Topics that apply only to specific Synergy HT models are preceded by a notice in italics for example Applies only to Synergy HT models with injectors L This icon calls attention to important information ...

Page 15: ...Stack Microplate Stacker The intended use of this instrument is dependent on the instrument s labeling If there is an IVD label then the instrument may be used for clinical research and development or other non clinical purposes If there is no such label then the instrument may only be used for research and development or for other non clinical purposes Quality Control It is considered good labora...

Page 16: ... adapter to connect primary power to the external power supply Use of a two prong adapter disconnects the utility ground creating a severe shock hazard Always connect the power cord directly to a three prong receptacle with a functional ground Warning Internal Voltage Always turn off the power switch and unplug the power supply before cleaning the outer surface of the instrument Warning Liquids Av...

Page 17: ...instrument to the recommended diluted sodium hypochlorite solution bleach for more than 20 minutes Prolonged contact may damage the instrument surfaces Be certain to rinse and thoroughly wipe all surfaces Caution Power Supply Only use the power supply shipped with the instrument Operate this power supply within the range of line voltages listed on it Caution Shipping Panel and Carrier Shipping Scr...

Page 18: ...tatic Discharge EN 61000 4 3 Radiated EM Fields EN 61000 4 4 Electrical Fast Transient Burst EN 61000 4 5 Surge Immunity EN 61000 4 6 Conducted Disturbances EN 61000 4 11 Voltage Dips Short Interruptions and Variations Directive 73 23 EEC Low Voltage Safety The system has been type tested by an independent testing laboratory and was found to meet the requirements of EC Directive 73 23 EEC for Low ...

Page 19: ...l area is likely to cause interference in which case the user will be required to correct the interference at his own expense Canadian Department of Communications Class A This digital apparatus does not exceed Class A limits for radio emissions from digital apparatus set out in the Radio Interference Regulations of the Canadian Department of Communications Le present appareil numerique n met pas ...

Page 20: ...courant alternatif Gleich und Wechselstrom Corriente continua y corriente alterna Corrente continua e corrente alternata Earth ground terminal Borne de terre Erde Betriebserde Borne de tierra Terra di funzionamento Protective conductor terminal Borne de terre de protection Schutzleiteranschluss Borne de tierra de protección Terra di protezione On Supply Marche alimentation Ein Verbindung mit dem N...

Page 21: ... ed intrappolarsi Warning hot surface Attention surface chaude Vorsicht heiße Oberfläche Precaución superficie caliente Attenzione superficie calda Separate collection for electrical and electronic equipment Les équipements électriques et électroniques font l objet d une collecte sélective Getrennte Sammlung von Elektro und Elektronikgeräten Recogida selectiva de aparatos eléctricos y electrónicos...

Page 22: ...xx Preface BioTek Instruments ...

Page 23: ...ntains information on contacting BioTek Instruments Inc for product support and service Synergy HT Multi Mode Microplate Reader 2 Features 3 Package Contents 4 Optional Accessories 5 Product Support Service 6 Technical Assistance Center TAC 6 Returning Instruments for Service Repair 6 Contacting BioTek for Applications Support 6 ...

Page 24: ...on flash lamp and a monochromator for wavelength selection The use of a xenon flash lamp allows for both UV and visible light absorbance measurements The monochromator provides wavelength selection from 200 to 999 nm in 1 nm increments The Synergy HT has a 4 Zone temperature control from 4 C over ambient to 50 C that ensures superior temperature uniformity necessary for kinetic assays Internal pla...

Page 25: ...ow noise PMT Excitation 300 to 650 nm 200 to 700 nm with T models Emission 300 to 700 nm Optional red extended PMT Excitation 300 to 650 nm 200 to 800 nm with T models Emission 300 to 800 nm Absorbance N range of 200 to 999 nm Absorbance OD range from 0 000 to 4 000 OD Low Medium High and Variable plate shaking speeds with adjustable durations All models read 6 12 24 48 96 and 384 well microplates...

Page 26: ...M Driver Software PN 7090204 Wrench PN 48576 Fluorescence lamp assembly PN 7080501 Note The part number for a replacement lamp is 7080500 Filter plugs 2 PN 7082073 also referred to as dummy filters Plastic storage bag and Velcro strips Time Resolved Fluorescence cartridge assembly PN 7090523 T models only Models with injectors SIAFRTD SIAFRTD CUSTOM an external dispense module PN 7090568 with the ...

Page 27: ... PN 7270512 Additional Fluorescence Filters contact BioTek for part numbers and availability Absorbance Liquid Test Solutions BioTek Wetting Agent Solution PN 7773002 BioTek QC Check Solution No 1 PN 7120779 25 ml or PN 7120782 125 ml BioTek Sodium Fluorescein Powder PN 98155 Models with injectors Dispense Module Liquid Test Solutions BioTek Green Test Dye Solution PN 7773003 BioTek Blue Test Dye ...

Page 28: ...rgy HT by selecting System Reader Control For troubleshooting assistance or instruments needing repair the specific steps that produce your problem and any error codes displayed in Gen5 KC4 see also Appendix C Error Codes Returning Instruments for Service Repair If you need to return an instrument to BioTek for service or repair please contact the TAC for a Return Materials Authorization RMA numbe...

Page 29: ...g Panel 11 3 Remove the Microplate Carrier Shipping Screw 13 4 Install the Fluorescence Lamp Assembly 14 5 Select an Appropriate Location 15 6 Connect the Power Supply 16 7 Unpack and Inspect the Dispense Module 17 8 Install the Dispense Module 20 9 Connect the Host Computer 24 10 Install the Software on the Host Computer 24 11 Turn on the Reader 25 12 Establish Communication 26 13 Run a System Te...

Page 30: ... the product s you have purchased Register online through BioTek s Customer Resource Center at www biotek com or by contacting BioTek Customer Care Once registered you can log into the Customer Resource Center and Register your product warranty Manage your equipment inventory Access documentation on your products Download user manuals and software Check the status of your instrument s service Trac...

Page 31: ...ct the packaging reader and accessories for shipping damage If the reader is damaged notify the carrier and your BioTek representative Keep the shipping boxes and the packaging materials for the carrier s inspection BioTek will arrange for repair or replacement of your reader immediately before the shipping related claim is settled Perform these steps to unpack and inspect the reader and accessori...

Page 32: ... and grasp the handles Carefully lift the reader out of the box and place it on a level surface Remove the protective plastic bag 4 Place all packing material back into the shipping box for reuse if the reader needs to be shipped again See Package Contents in Chapter 1 for assistance with identifying the contents of the accessories box Accessories box Inner shipping box Vertical supports Figure 1 ...

Page 33: ...ing panel to the bottom of the reader See Figure 2 on the next page 3 Carefully set the reader upright 4 Locate the supplied plastic tool storage pocket Place the screws and washers inside the bag Use the supplied Velcro strips to attach the pocket to the back of the reader for storage Do not block any air vents See Figure 3 on the next page 5 Place the panel back into the inner shipping box for s...

Page 34: ...12 Chapter 2 Installation BioTek Instruments Inc Figure 2 Removing the shipping panel Figure 3 Storage pocket on the rear of the instrument wrench carrier shipping screw and warning tag shown ...

Page 35: ...e reader 2 Using the supplied wrench remove the carrier shipping screw with its o ring and warning tag 3 Place the wrench screw o ring and tag in the plastic tool storage bag that you attached to the back of the reader Figure 4 Removing the microplate carrier shipping screw L Important Replace the microplate carrier shipping screw before repackaging the Synergy HT for shipment Please contact BioTe...

Page 36: ...embly 1 Locate the lamp assembly in the accessories box The lamp is attached to a metal bracket that also holds a condenser lens and a heat absorber Two cables are attached to the back of the lamp 2 Open the hinged door on the front of the reader by pressing on its lower left and right corners The lamp compartment is on the far left 3 Orient the lamp assembly as shown below Slide the assembly all ...

Page 37: ...ive humidity range for this reader is from 10 to 85 non condensing Excessive ambient light Bright sunlight or strong incandescent light may affect the reader s optics and readings reducing its linear performance range Dust Readings may be affected by extraneous particles such as dust in the microplate wells A clean work area is necessary to ensure accurate readings Note If you will be installing B...

Page 38: ... of a two prong adapter disconnects the utility ground creating a severe shock hazard Always connect the power cord directly to an appropriate receptacle with a functional ground Perform these steps to connect the power supply 1 Connect the power cord to the external power supply 2 Locate the power inlet on the rear of the reader 3 Plug the rounded end of the power supply s line cord into the powe...

Page 39: ...rocess inspect the packaging module and accessories for shipping damage If the reader is damaged notify the carrier and your BioTek representative Keep the shipping boxes and the packaging materials for the carrier s inspection BioTek will arrange for repair or replacement of your reader immediately before the shipping related claim is settled Perform these steps to unpack and inspect the dispense...

Page 40: ...ing box 3 Open the accessories box Remove and identify its contents see Figure 9 on the next page 2 inlet tubes packaged in plastic cylinders 4 outlet tubes packaged in plastic bags PN 7082120 2 syringes packaged in boxes 1 priming plate 2 reagent bottles 1 injector tip priming trough small plastic cup 1 plastic tool storage bag with Velcro strips 2 metal thumbscrews 1 stylus wire packaged in a sm...

Page 41: ...ule 19 Synergy HT Operator s Manual Figure 9 Unpacking the dispense module s accessories Accessories box Bottom foam end cap with cutouts Syringes 2 Dispense module cover Outlet tubes 4 Inlet tubes 2 Top foam end cap Dispense module cable ...

Page 42: ... 4 Place the nylon screws and the plastic fitting covers in the plastic tool storage bag Use the supplied Velcro strips to attach the bag to the rear panel of the dispense module 5 Remove the two inlet tubes from their protective plastic canisters 6 Identify the two circular syringe valves on the dispense module Each is labeled with a left pointing arrow See Figure 12 on the next page When install...

Page 43: ...odule 21 Synergy HT Operator s Manual Figures 10 and 11 Possible locations for the dispense module to the left or on top of the instrument Figure 12 Initial setup of the dispense module Inlet tubes Outlet tubes Syringe valves ...

Page 44: ...ertically with the threaded end at the top and the knurled steel end at the bottom Screw the threaded end of the syringe into the bottom of the syringe valve Finger tighten only Carefully pull down the knurled steel end of the syringe until it is resting inside the hole in the bracket Pass a metal thumbscrew up through this hole and thread it into the bottom of the syringe Hold the syringe from ro...

Page 45: ...e Dispenser Port on the rear panel of the Synergy HT Figure 14 Dispense module connected to the reader rear view 15 Locate the injector tip cleaning stylus packaged in a small plastic cylinder Attach the cylinder to the back of the dispense module for storage One end of the cable connected to the port on the side of the dispense module The other end connected to the reader s Dispenser Port ...

Page 46: ...ropriate port on the reader see photo below and the other end to the appropriate port on the host computer Figure 15 RS 232 serial and USB ports on the rear panel injector model shown 10 Install the Software on the Host Computer The Synergy HT is controlled by BioTek s Gen5 or KC4 software running on a host computer There is a certain sequence of events that must be followed to ensure that the sof...

Page 47: ... s Manual 11 Turn on the Reader Locate the power switch on the front panel and turn on the Synergy HT The reader will automatically initiate a System Test and eject the microplate carrier Figure 16 Carrier eject button top and power ON OFF switch ...

Page 48: ... connected If using the USB cable the information can be found via the Windows Control Panel under Ports in the Hardware Device Manager area of System Properties e g USB Serial Port COM5 7 Click the Test Comm button Gen5 will attempt to communicate with the reader If the communication attempt is successful return to Gen5 s main screen If the communication attempt is not successful try the followin...

Page 49: ...Ports in the Hardware Device Manager area of System Properties e g USB Serial Port COM5 6 Click the Current Reader button KC4 will attempt to communicate with the reader If a message appears stating that the software wavelength table does not match the reader filter table this means that communication has been established and KC4 just needs to update its wavelength table Click Yes If the communica...

Page 50: ...ear Scroll down toward the bottom the text should read SYSTEM TEST PASS You may wish to print the report and store it with your Installation records The software stores system test information in its database you can retrieve it at any time If an error code is returned turn to Appendix C Error Codes and look up the code If the problem is something you can fix do so now and run another System Test ...

Page 51: ...trough in the left rear pocket of the carrier Place the priming plate on the carrier Figure 17 Installing the tip priming trough and priming plate on the microplate carrier 2 Fill the two reagent bottles with distilled or deionized water Place the bottles in their holders and place the holders directly in front of the syringes Insert the inlet tubes into the bottles The dispense module s setup sho...

Page 52: ... tubing and dispense into the priming plate Examine the fittings no leaks should be detected If leaks are detected tighten all fittings and repeat the prime If leaks are still detected contact BioTek s Technical Assistance Center 6 When the prime finishes set Volume to 2000 µl and click Purge to clear the fluid lines 7 Set Dispenser to 2 and repeat steps 5 and 6 8 When finished remove and empty th...

Page 53: ...t this chapter If you suspect that problems occurred during shipment if you received the reader back from BioTek following service or repair and or if regulatory requirements dictate that Operational Performance Qualification is necessary turn to Chapter 4 Instrument Qualification now to learn about BioTek s recommended OQ PQ procedures for the Synergy HT Note An Installation Operational Performan...

Page 54: ... guidance Replace the microplate carrier shipping screw and the shipping panel before repackaging the reader for shipment Please contact BioTek if you have misplaced either of these items If you need to ship the Synergy HT and or the dispense module to BioTek for service or repair be sure to use the original packaging materials Other forms of commercially available packaging are not recommended an...

Page 55: ...d in the plastic bag attached to the back of the module Set the module aside for the moment 4 If you have not already done so retract the microplate carrier and then turn off and unplug the reader 5 Remove the lamp assembly and pack it in bubble wrap see p 14 6 Replace the microplate carrier shipping screw see p 13 7 Tip the reader onto its back feet Attach the shipping panel to the bottom of the ...

Page 56: ...ear plastic fitting covers to the fittings of the outlet tubes Place the tubes in a plastic bag 4 Place the dispense module inside the inner shipping box Slide the cardboard shipping insert down around the module Pack the reagent bottle holders in bubble wrap and place them on top of the module Seal the box with tape 5 Locate the original accessories shipping box and foam end caps Place the bottom...

Page 57: ...n Filter Wheels 38 Installing the Time Resolved Fluorescence Cartridge 41 Configuring the System for Luminescence Measurements 42 The External Dispense Module 43 Gen5 Software 46 Viewing Updating the Filter and Wavelengths Tables 46 Creating Protocols and Experiments 47 Controlling the Dispense Module 50 KC4 Software 52 Viewing Updating the Filter and Wavelengths Tables 52 Creating Protocols 53 Re...

Page 58: ...te securely in place The microplate loading door helps to ensure a light impermeable measurement chamber When a plate read is initiated the carrier slides into the measurement chamber and then moves in the X and Y axes to align each microwell with the top or bottom fluorescence probe or bottom absorbance probe as specified in the Gen5 procedure or KC4 protocol When the read is complete the plate c...

Page 59: ...nce one for absorbance and time resolved fluorescence Standard Fluorescence The 20 watt tungsten halogen lamp s life is rated at an average of 1000 hours and it is user replaceable The intensity of the bulb will slowly drop over time until the instrument s run time self check detects a low lamp current signal and Gen5 or KC4 displays an error message The lamp PN 7080500 should be replaced at this ...

Page 60: ...T Each filter wheel is labeled EX or EM and can contain up to four filters and or black plugs A filter can be used in either wheel but it must be oriented properly as described below Each filter and plug is held securely in place with a C clip filter retainer Note Each filter has its wavelength and bandpass values printed on its side with an arrow to indicate the proper direction of light through ...

Page 61: ...s follows EX Excitation EM Emission TR Time Resolved Cartridge see page 41 Filter direction within a filter wheel is important and the direction differs depending on the filter wheel There is a diagram on the inside of the front panel door indicating this Each filter is marked with an arrow indicating the proper direction of light Refer to the figures on the previous page for proper filter orienta...

Page 62: ...our finger covered with the lens paper gently push against the filter and its C clip retainer until they pop out To replace a filter or plug 1 Hold the metal bracket with the filter wheel facing up 2 Properly orient the filter or plug see page 38 and then drop it into the desired filter wheel location 3 Using your fingers squeeze the sides of the C clip filter retainer and then insert it into the ...

Page 63: ... of the hinged door on the front of the instrument Observe the two thumbscrews within the compartment The left thumbscrew holds the Excitation filter wheel in place See the figure on page 37 3 Remove the left thumbscrew and slide the filter wheel s supporting metal bracket straight out of the compartment 4 Slide the TR cartridge into the compartment and replace the thumbscrew Close the front door ...

Page 64: ...he Emission filter wheel and examine its contents ensure that there is an empty location If you made any changes to either filter wheel you must update Gen5 s or KC4 s filter table Select PLUG to indicate the presence of a plug and HOLE to indicate an empty location Click Send Values or Filters to download the information to the reader Updating Gen5 s filter table for complete instructions see pag...

Page 65: ...les d Inlet tubes transport fluid from the supply vessels to the syringes These tubes are short pieces of opaque PTFE Teflon tubing connected to stainless steel probes on one end and threaded fittings on the other end e Three way valves switch the syringe flow from the inlet tubes to the outlet tubes f Outlet tubes transport fluid from the syringes into the instrument through the tubing ports on t...

Page 66: ...instrument to the two injectors As shown below both injectors are positioned directly above the bottom fluorescence optical probe Syringe 1 Syringe 2 Bottom probe Figure 24 Close up view of the injectors inside the instrument Note The tubing and injectors should be cleaned at least quarterly See Chapter 5 Preventive Maintenance for more information ...

Page 67: ...ctivities are controlled via Gen5 see page 50 or KC4 see page 56 Both types of primes require a fluid reservoir to be present on the microplate carrier The priming plate is about the same size as a standard microplate and is placed on the microplate carrier for a Prime operation to prime the dispense system with fluid The tip priming trough is a small removable priming cup located in the left rear...

Page 68: ...To view this table in Gen5 select System Reader Configuration highlight the Synergy reader and click View Modify Click Setup and then click the Fluorescence Luminescence tab Regarding the Absorbance Wavelengths table The Synergy HT performs absorbance reads in the range of 200 to 999 nm Click the Absorbance tab to specify and calibrate 6 wavelengths to be made available as default selections withi...

Page 69: ...the data retrieved from the reader At a minimum a protocol must specify the Procedure for the assay you wish to run After creating a protocol create an Experiment that references the protocol You ll run the experiment to read plates and analyze the data Figure 26 Defining the Procedure within a Gen5 Protocol Figure 27 An Experiment containing measurement data based on a pre defined protocol ...

Page 70: ...ding the plate and more Click the Validate button to verify that the reader supports the defined steps and then click OK Tips Add a Dispense step to define the volume and rate at which fluid will be dispensed and from which dispenser Add a Read step to specify the detection method and filter sets or wavelength values enable time resolved fluorescence and set the Top Probe Vertical Offset value To ...

Page 71: ...how to create a simple Experiment and then read a plate in Gen5 See Gen5 s Help system for complete instructions To create an Experiment and read a plate using Gen5 1 Select File New Experiment 2 Select the desired protocol and click OK 3 Highlight a plate in the menu tree and select Plate Read The Plate Reading dialog will appear 4 Click READ The door will open and the carrier will extend if it i...

Page 72: ...e button The syringe drive will move to its home position and its sensors will be verified Upon successful completion the Initialized field should show Yes Prime Utility Before running an experiment with a Dispense step the dispense module and its associated tubing must be primed with the fluid to be used Gen5 provides a special utility for this task To prime the dispense module 1 Fill the supply ...

Page 73: ...unctions for maintenance and calibration purposes If a syringe needs to be installed or replaced it must first be moved to its Maintenance Position To do this using Gen5 1 In Gen5 select System Reader Control Synergy Com and click the Dispenser tab 2 Select the appropriate Dispenser number 1 or 2 associated with the syringe 3 Click Maintenance The syringe plunger will move to its furthest from hom...

Page 74: ...C4 by selecting System Reader and clicking the Filters Wavelengths button Click the Fluorescence Luminescence tab to view the Excitation and Emission filter wheel information retrieved from the Synergy Regarding the Absorbance Wavelengths table The Synergy HT performs absorbance reads in the range of 200 to 999 nm Click the Absorbance tab to specify and calibrate 6 wave lengths to be made availabl...

Page 75: ...h to run Figure 29 KC4 s Reading parameters dialog software version 3 4 The instructions on the following pages briefly describe how to create protocols in KC4 Be sure to refer to KC4 s Help system and User s Guide for complete instructions Note KC4 software versions 3 4 and greater offer a Multi Mode option within the Reading Parameters dialog This option allows you to define more than one detect...

Page 76: ...nd give the protocol an identifying name To create a Fluorescence or Luminescence non Dispense protocol in KC4 1 Select Data New Plate If prompted to select a protocol select Empty Protocol and click OK If not prompted select Protocol New 2 Select Protocol Reading The Reading parameters dialog will appear 3 Set Detection Method to Fluorescence or Luminescence 4 Select a Reading Type of Endpoint or...

Page 77: ...button The Read Dispense dialog will appear Select a Read Mode of Well or Plate Click the Dispense Shake and Read buttons to add steps to the protocol When finished click OK to return to the Reading parameters dialog 10 Click OK to verify the parameters and return to the main screen 11 Select Protocol Save As and give the protocol an identifying name Reading Plates To read a plate using KC4 1 Sele...

Page 78: ...e syringe drive will move to its home position and its sensors will be verified Upon successful completion the Status should show Initialized 4 Repeat for the other syringe Dispenser 2 Prime Utility Before running a Read Dispense protocol the dispense module and its associated tubing must be primed with the fluid to be used KC4 provides a special utility for this task To prime the dispense module ...

Page 79: ... and must be periodically emptied and cleaned by the user If the Tip Priming Trough overflows you should clean the microplate carrier and possibly the internal surface beneath the carrier See Chapter 5 Preventive Maintenance for instructions See also Appendix A for decontamination instructions KC4 must be told when the trough has been emptied To empty the tip priming trough 1 If the microplate car...

Page 80: ...utton and then click the appropriate Dispenser tab 1 or 2 2 Click Move Syringe to maintenance position The syringe plunger will move to its furthest from home position The syringe can then be disconnected form the drive bracket and unscrewed from the valve See Install Dispense Module Components in Chapter 2 Installation for information on installing removing the syringes 3 Click OK to close the di...

Page 81: ...uracy especially with U and V bottom polyvinyl microplates Check for this by reading an empty microplate Dual wavelength readings can eliminate this problem or bring the variation in density readings to within acceptable limits for most measurements Inaccuracy in pipetting has a large effect on measurements especially if smaller volumes of liquid are used For best results use at least 100 l per we...

Page 82: ...geways Flushing the tubing at the end of each day letting the DI water soak them overnight and then purging the lines at the beginning of each day ensures optimal performance of the dispense system See Chapter 5 Preventive Maintenance for more information For models with injectors When dispensing volumes less than or equal to 20 L well we recommend specifying a tip prime volume that is equal to th...

Page 83: ...ngoing performance of the Synergy HT and the external dispense module if used Overview 62 IQ OQ PQ 62 Recommended Qualification Schedule 64 System Test 65 Absorbance Plate Test 70 Absorbance Liquid Tests 80 Fluorescence Tests 89 Dispense Module Tests 102 Dispense Accuracy Precision Tests Worksheet 121 ...

Page 84: ...n Operational Qualification Performance Qualification and Preventive Maintenance procedures Microsoft Excel spreadsheets are provided for performing the calculations and checklists data sheets and logbooks are provided for recording results Contact your local BioTek dealer for more information IQ OQ PQ Installation Qualification confirms that the reader and its components have been supplied as ord...

Page 85: ...with those from the routine Performance Qualification tests and previous OQ tests to monitor for trends The successful completion of the OQ procedure in combination with results that are comparable to previous PQ and OQ tests confirms that the equipment is operating according to specification initially and over time Performance Qualification confirms that the reader consistently meets the requirem...

Page 86: ...quid Test 2 9 9 Liquid Test 3 9 9 Fluorescence Tests Corners Test 9 9 Sensitivity Linearity Tests 9 9 Tests for Injector Models Injector System Test 9 Dispense Accuracy and Precision Tests 9 9 Regarding Liquid Tests 1 and 2 If you have an Absorbance Test Plate run Liquid Test 1 If you do not have an Absorbance Test Plate run Liquid Test 2 Liquid Test 3 is optional it is provided for sites requirin...

Page 87: ...st report generated via Gen5 or KC4 The report also contains the reader s serial number and the part number and version number of the basecode software When the instrument is turned on the System Test automatically runs but no report is generated To run the test and generate a report you must use Gen5 or KC4 The absorbance measurement system is checked using the six wavelengths specified in the re...

Page 88: ...tain the error code that was generated by the failure 5 When the test is complete a dialog will appear requesting additional information Enter the information if desired and then click OK 6 The test report will appear Scroll down toward the bottom of the report it will show either SYSTEM TEST PASS or SYSTEM TEST FAIL ERROR error code DETECTED 7 Print the report if desired A sample test report is s...

Page 89: ...90 1 counts PASS 750V noise 66 counts 500V measurement 4 6 counts 500V noise 3 counts Lambda 200 Gain 1 80 Resets 1 Channel Ref 1 Air 14235 39595 Dark 9869 9889 Delta 4366 29706 Lambda 340 Gain 1 95 Resets 2 Channel Ref 1 Air 12954 39951 Dark 9871 9885 Delta 3083 30066 Lambda 405 Gain 1 61 Resets 2 Channel Ref 1 Air 12755 38902 Dark 9871 9883 Delta 2884 29019 Lambda 550 Gain 1 63 Resets 1 Channel ...

Page 90: ...wer Right Corner x 1044 y 5888 Upper Right Corner x 1056 y 356 Delta 1 9732 9720 12 Delta 2 1056 1044 12 Delta 3 356 360 4 Delta 4 5888 5896 8 PROBE BOTTOM Upper Left Corner x 9724 y 1844 Lower Left Corner x 9716 y 7376 Lower Right Corner x 1040 y 7380 Upper Right Corner x 1040 y 1852 Delta 1 9724 9716 8 Delta 2 1040 1040 0 Delta 3 1852 1844 8 Delta 4 7380 7376 4 PROBE ABSORB Upper Left Corner x 1...

Page 91: ...rt In the U S In Europe BioTek Instruments Inc BioTek Instruments GmbH Tel 800 242 4685 Tel 49 0 7136 9680 Fax 802 655 3399 Fax 49 0 7136 968 111 All Others Tel 802 655 4040 Fax 802 655 3399 email TAC biotek com Product support center www biotek com service Figure 30 3 Sample output for the System Test Sheet 3 of 3 The format varies depending on the software used ...

Page 92: ... being clipped and the reader may be out of alignment Accuracy Linearity The Test Plate contains neutral density glass filters of known OD values at several wavelengths Actual measurements are compared against the expected values provided in the Test Plate s Standards Certificate Since there are several filters with differing OD values the accuracy across a range of ODs can be established Once it ...

Page 93: ...Tek autoreader The following calibration data has been recorded by a N I S T traceable spectrophotometer WAVELENGTH nm Well 405nm 450nm 490nm 550nm 620nm 630nm 690nm 750nm C1 0 147 0 140 0 135 0 130 0 136 0 136 0 127 0 134 E2 0 618 0 575 0 574 0 568 0 573 0 568 0 485 0 434 G3 1 133 1 052 1 051 1 040 1 050 1 040 0 881 0 783 H6 1 701 1 578 1 577 1 560 1 575 1 560 1 323 1 179 F5 2 279 2 024 1 976 1 9...

Page 94: ...h combination If you need to change the wavelength values click the Wavelength List button Click Gen5 s Help button for assistance 7 Select the number of Peak Wavelength tests to run 1 to 4 based on the number of peak wavelength values provided on the Peak Wavelength Certificate 8 Enter the Expected Peak value s from the 2 4 nm Spectral Bandpass column of the Peak Wavelength Certificate Select 1 4...

Page 95: ... for your use of the reader 5 Optional Enter any Comments 6 Click Start Test 7 Place the Test Plate in the microplate carrier so that well A1 is in the left rear corner of the carrier as you are facing the carrier 8 Click OK to run the test 9 When the test completes the results report will appear Scroll down through the report every result should show PASS See page 78 for information on results an...

Page 96: ... Certificate Note If the certificate specifies multiple peak wavelength values you will need to run an independent test using spectral scan reads KC4 supports the entry of only one Peak Wavelength value and it must be in the range of 580 to 590 nm See the special instructions on the next page 7 Review all of the values you entered and then click OK to save the data The information you just entered...

Page 97: ...st was performed Performing Peak Absorbance Tests Using Spectral Scan Reads To create and run a KC4 spectral scan protocol to perform an independent Peak Absorbance Test Note Create one protocol per Peak Wavelength value to be tested 1 Open the Reading Parameters dialog and select the Spectrum Reading Type and the Absorbance Detection Method 2 Set the Start and Stop wavelength values to define the...

Page 98: ... 618 1 133 1 701 2 279 2 945 Min Limit 0 124 0 586 1 090 1 647 2 168 2 807 Max Limit 0 170 0 650 1 176 1 755 2 390 3 083 Read 1 0 144 0 615 1 128 1 696 2 284 2 908 Result PASS PASS PASS PASS PASS PASS Repeatability Results Wells C1 E2 G3 H6 F5 D4 Read 1 0 144 0 615 1 128 1 696 2 284 2 908 Min Limit 0 138 0 604 1 112 1 674 2 210 2 816 Max Limit 0 150 0 626 1 144 1 718 2 358 3 000 Read 2 0 144 0 615...

Page 99: ...PASS PASS Reviewed Approved By _______________________________________ Date ________________ For Technical Support In the U S In Europe BioTek Instruments Inc BioTek Instruments GmbH Tel 800 242 4685 Tel 49 0 7136 9680 Fax 802 655 3399 Fax 49 0 7136 968 111 All Others Tel 802 655 4040 Fax 802 655 3399 email TAC biotek com Product support center www biotek com service Figure 32 2 Sample output for ...

Page 100: ...e Test Plate is within its calibration certification period The calibration sticker is affixed directly to the plate If it is out of date contact BioTek to schedule a recertification Check the microplate carrier to ensure it is clear of debris Alignment This portion of the test measures the alignment of the microplate carrier with the optical path A reading greater than 0 015 OD represents an out ...

Page 101: ...r of debris Linearity of the optical density readings is confirmed by default if the optical density readings are accurate To further verify this perform a regression analysis on the Test Plate OD values in a program such as Microsoft Excel as follows 1 Launch Excel 2 Create a spreadsheet and label one column Assigned and the next column Observed 3 Enter the Assigned OD data for each glass filter ...

Page 102: ...than those attainable with the Absorbance Test Plate This test is optional because the reader has good front end linearity throughout its wavelength range For Liquid Tests 1 and 2 the tester is instructed to prepare the stock dye solution described on the next page The purpose of the formulation is to create a solution that absorbs light at 2 0 OD full strength when dispensed at 200 L in a flat bo...

Page 103: ...ion A Required Materials Deionized water FD C Yellow No 5 dye powder typically 90 pure Tween 20 polyoxyethylene 20 sorbitan monolaurate or BioTek wetting agent PN 7773002 Precision balance with capacity of 100 g minimum and readability of 0 001 g Weigh boat 1 liter volumetric flask Procedure 1 Weigh out 0 092 g of FD C yellow No 5 dye powder into a weigh boat 2 Rinse the contents into a 1 liter vo...

Page 104: ...tion before reading the plate 4 Using Gen5 or KC4 read the microplate five times at 405 nm using the Normal read mode single wavelength no blanking Save the data after each read Normal plate position 5 Without delay rotate the microplate 180 degrees so that well A1 is in the H12 position Read the plate five more times saving the data after each read Turnaround plate position 6 Print out the ten se...

Page 105: ...0 05 solution of deionized water and Tween 20 Prepare the Dilutions Refer to the table below and Create a percentage dilution series beginning with 100 of the original concentrated stock solution A or B in the first tube 90 of the original solution in the second tube 80 in the third tube all the way to 10 in the tenth tube Dilute using the 0 05 solution of deionized water and Tween 20 Table 2 Test...

Page 106: ...he five sets of Delta OD data or export them to an Excel spreadsheet Data Reduction for Linearity 3 Calculate the mean absorbance for each well and average the means for each concentration 4 Perform a regression analysis on the data to determine if there is adequate linearity For example using Microsoft Excel In a spreadsheet create two columns labeled X and Y Reference Table 2 on the previous pag...

Page 107: ...05 OD from 2 000 OD to 3 000 OD Alignment Test 1 Using the plate prepared for the Linearity Test on the previous page conduct a Turnaround test by reading the plate five times with the A1 well in the H12 position Save the data after each read This test results in values for the four corner wells that can be used to determine alignment 2 Calculate the means of the wells A1 and H1 in the Normal plat...

Page 108: ... have an optical density of approximately 0 700 to 1 000 This value is not critical but it should be within this range If low adjust up by adding β NADH powder until the solution is at least at the lower end of this range Do not adjust if slightly high Buffer Solution Sigma PBS Solution Deionized water Phosphate Buffered Saline PBS pH 7 2 7 6 Sigma tablets P4417 or equivalent β NADH Powder β Nicot...

Page 109: ...fter each read 5 Print out the five sets of raw data or export them to an Excel spreadsheet Data Reduction for Repeatability 6 For each well calculate the mean and standard deviation of the five readings 7 For each mean calculated in step 6 calculate the allowed deviation using the repeatability specification for a 96 well plate of 1 0 0 005 OD Mean x 0 010 0 005 8 For each well compare the standa...

Page 110: ...s Enter 0 50 0 75 and 1 00 and label the column as the X values Select Tools Data Analysis Regression to open the Regression dialog Set Input Y Range to reference the above mentioned Y values and set Input X Range to reference the above mentioned X values Click OK to perform the analysis the results of which will be output in a separate sheet Note If the Data Analysis command is not available on t...

Page 111: ... distinguishable from the mean of wells with pure diluent The Linearity Test verifies that the system is linear that is signal changes proportionally with changes in concentration Proving that the system is linear allows the Sensitivity Test to be run on two points instead of using serial dilutions L Important The tests presented in this section require specific microplates solutions and EX EM fil...

Page 112: ...odium Fluorescein Powder 1 mg vial BioTek PN 98155 Bottom optics A clean Hellma Quartz 96 well titration plate Mfr 730 009 QG or equivalent such as the 96 well glass bottom Greiner SensoPlate Mfr 655892 Top optics A new clean 96 well solid black microplate such as Corning Costar Mfr 3915 The Hellma Quartz plate described above can also be used Excitation filter 485 20 nm and Emission filter 528 20...

Page 113: ...ium Borate solution does not require further preparation proceed to step 2 If you are using PBS prepare the solution now Optional but recommended Using a 0 45 micron filter filter 200 mL of deionized or distilled water Follow the manufacturer s instructions on the PBS packaging to create 200 mL dissolving the necessary amount of PBS into the filtered water Stir the solution preferably using a stir...

Page 114: ... FI_T prt protocol 5 Save and or print the measurement data 6 Calculate and analyze the results as described below Results Analysis Corners Test 1 Calculate the Mean of the twelve wells containing the 3 3 nM SF test solution A1 A3 A10 A12 H1 H3 H10 H12 2 Calculate the Standard Deviation of the same twelve wells 3 Calculate the CV Standard Deviation Mean 100 The CV must be less than 3 0 to pass Sen...

Page 115: ... the Excitation Emission filters clean Are they in the proper locations and in the proper orientation in the filter wheels Are you using new clean plates We suggest you re run the test with a new clean microplate For the bottom optics test if the base of a plate is touched clean the entire base with alcohol 95 ethanol and then wipe with a lint free cloth Before placing a plate in the instrument bl...

Page 116: ...he 1 nM SF solution into wells C1 F1 Discard the tips Pipette 150 µL of the 1 nM SF solution into wells C2 F2 Mix the wells using the pipette Do not discard the tips Aspirate 150 µL from wells C2 F2 and dispense into wells C3 F3 Mix the wells using the pipette Do not discard the tips Aspirate 150 µL from wells C3 F3 and dispense into wells C4 F4 Mix the wells using the pipette Do not discard the t...

Page 117: ...ad Wells Corners Read step All wells Sensitivity Read step Wells C1 F12 Filters EX 485 20 nm EM 528 20 nm Optics Position Bottom Sensitivity Corners Read step 80 Sensitivity Linearity Read step 100 Delay After Plate Movement 350 msec Measurements Per Data Point 40 Protocol Name Synergy HT FI_T prt Parameter Setting Plate Type Costar 96 black opaque Mfr 3915 Read Wells Corners Read step All wells S...

Page 118: ...tom Sensitivity Filter Set 1 Corners 80 Filter Set 2 Sensitivity Linearity 100 Nb Samples Per Well Both filter sets 40 Delay Before Sampling Both filter sets 350 msec Plate Type Greiner SensoPlate Mfr 655892 Note You may need to add this Plate Format in KC4 see the dimensions on the next page Read Wells A1 H12 Protocol Name Synergy HT FI_T prt Parameter Setting Reading Type Endpoint Detection Meth...

Page 119: ...Plate Format Dimensions To create a plate format in KC4 for the Greiner SensoPlate select System Plate Formats click New and add the information shown below Once the plate format is created you can reference it in a protocol s Reading Parameters dialog ...

Page 120: ...oil Various beakers graduated cylinders and pipettes Optional but recommended 0 45 micron filter Gen5 or KC4 Synergy HT FI_MUB prt protocol described on page 101 Test Solutions 1 Prepare the buffer CBB solution Optional but recommended Using a 0 45 micron filter filter 200 mL of deionized or distilled water Open and dissolve the contents of 2 CBB capsules do not dissolve the outer gelatin capsule ...

Page 121: ... Refer to the Pipette Map on page 100 and pipette the solutions into a clean 96 well solid black plate 4 Read the plate using the Synergy HT FI_MUB prt protocol 5 Save and or print the measurement data 6 Calculate and analyze the results as described below Results Analysis Sensitivity Test 1 Calculate the Mean and Standard Deviation for the buffer wells C10 F12 2 Calculate the Mean for the 17 6 ng...

Page 122: ...mL 100 nM MUB solution into wells C1 F1 Discard the tips Pipette 150 µL of the 17 6 ng mL solution into wells C2 F2 Mix the wells using the pipette Do not discard the tips Aspirate 150 µL from wells C2 F2 and dispense it into wells C3 F3 Mix the wells using the pipette Do not discard the tips Aspirate 150 µL from wells C3 F3 and dispense it into wells C4 F4 Mix the wells using the pipette Do not d...

Page 123: ...Setting Plate Type Costar 96 black opaque 3915 Read Wells Wells C1 to F12 Detection Method Fluorescence Read Type Endpoint Filters EX 360 40 nm EM 460 40 nm Optics Position Top Sensitivity 90 Delay After Plate Movement 350 msec Measurements Per Data Point 40 Top Probe Vertical Offset 1 00 mm KC4 USERS Protocol Name Synergy HT FI_MUB prt Parameter Setting Reading Type Endpoint Detection Method Fluo...

Page 124: ...tion and one 96 well microplate per injector to test the three different volumes The balance is tared with the empty plate and then the 80 µL dispense is performed for columns 1 4 The fluid is weighed and the balance is tared again with the plate on the balance This process is repeated for the 20 µL and 5 µL dispenses It is assumed that the solutions used are at room temperature A precision balanc...

Page 125: ...ier in this chapter Shaker if the absorbance reader does not support shaking Precision balance with capacity of 100 g minimum and readability of 0 001 g 50 200 l hand pipette and disposable tips Deionized water Supply bottles 250 mL beaker New 96 well flat bottom microplates BioTek s Green Test Dye Solution PN 7773003 undiluted or one of the alternate test solutions provided on the next page 100 m...

Page 126: ...Dye Solution PN 7773003 prepare a green test dye solution using one of the following methods Using BioTek s Blue and Yellow Concentrate Dye Solutions Ingredient Quantity Concentrate Blue Dye Solution PN 7773001 125 mL 4 0 mL QC Yellow Solution PN 7120782 125 mL 5 0 mL Deionized water 90 0 mL Using FD C Blue and Yellow Dye Powder Ingredient Quantity per Liter FD C Blue No 1 0 200 grams FD C Yellow ...

Page 127: ...he Results Analysis calculations See the Gen5 Help System for information on the Power Export Builder Test Setup KC4 Using KC4 create three Dispense protocols for the Synergy HT If you are using a BioTek absorbance reader create two additional Read protocols See page 115 for instructions If you are not using a BioTek absorbance reader prepare your reader to perform two reads with the following cha...

Page 128: ...nsers with 2000 µL of the solution When finished remove the priming plate from the carrier 4 Create a new experiment based on the Synergy HT Dispense 1 prt file described on page 110 5 Place a new 96 well microplate on the balance and tare the balance 6 Place the plate on the microplate carrier Important Gen5 will provide prompts and instructions for processing the plates follow the steps carefull...

Page 129: ...15 second shake the 80 µL read at 405 750 nm and the 20 and 5 µL read at 630 750 nm 9 When processing is complete select File Save As and save the experiment using an identifying file name 10 Repeat steps 4 9 using the Synergy HT Dispenser 2 prt protocol 11 See page 109 for instructions for analyzing the results 12 When all tests are complete prime both dispensers with at least 5000 µL of deionize...

Page 130: ...eps carefully When each dispense protocol is finished you will weigh the plate record the weight and tare the balance 3 Place a new 96 well microplate on the balance and tare the balance 4 Place the plate on the carrier and run the 80 μl Dispense protocol created for the dispenser under test 5 Weigh the plate Record the 80 µL dispense weight and then tare the balance 6 Place the plate on the carri...

Page 131: ...e KC4 Read protocols as described on page 117 the CV is calculated automatically With the Data file open select the Delta OD data set and click the Statistics button For each volume dispensed 80 20 5 µL for each dispenser 1 2 Calculate the Standard Deviation of the 32 wells Calculate the Mean of the 32 wells Calculate the CV Standard Deviation Mean x 100 Calculate the Accuracy Error Actual Weight ...

Page 132: ...lecting System Reader Configuration and add configure the Synergy if it is not already there 2 Select File New Protocol A menu tree will appear To edit a protocol category double click its branch in the tree as shown below Figure 33 Editing the Procedure in a new Protocol 3 Perform the steps in the following three sections to define the Procedure customize the Plate Layout and add Data Reduction s...

Page 133: ...lta OD Read columns 5 12 at 630 750 nm calculate the Delta OD The detailed procedure is described on the next page To add a step to the procedure click the appropriate button on the left side of the Procedure dialog see Figure 33 on page 110 and define the required parameters The comments suggested for use with the Plate Out In steps are optional but they may be useful for the person running the t...

Page 134: ...the weight TARE the balance Place the plate back on the carrier Click OK to continue 5 Dispense Dispenser select 1 or 2 depending on the protocol Dispense to wells A9 H12 Tip prime before this dispense step 5 µL Dispense 5 µL at rate 225 µL sec 6 Plate Out In Suggested comment Weigh the plate 5 ul test RECORD the weight PIPETTE 150 ul well of DI water into all 12 columns Place the plate back on th...

Page 135: ...ght wells A9 to H12 7 Click OK to save the changes and close the dialog Tip After running the experiment view the Statistics for each Delta OD Data Set to view the calculations Add Data Reduction Steps Each Read step is performed using two wavelengths so you ll need to create two data reduction steps to calculate the Delta OD values 1 In the protocol open the Data Reduction dialog and click the Tr...

Page 136: ...he 20 and 5 µL Read step at 630 nm DS2 set to the 20 and 5 µL Read step at 750 nm New Data Set Name resembling Delta OD 20 and 5 ul_Disp Formula DS1 DS2 applied to wells A5 to H12 11 When you re finished the Data Reduction Steps list will show two Delta OD transformations 12 Click OK to close the Data Reduction dialog Select File Save As and give the protocol an identifying file name such as Syner...

Page 137: ...he following set of steps three times to create six protocols for the 80 20 and 5 μl tests for Dispenser 1 and Dispenser 2 1 If a Data file is open close it now Data Close 2 Select Protocol New and then Protocol Reading The Reading Parameters dialog opens 3 Select the Dispense box and set the Reading Type to Endpoint 4 Click the Read Dispense button The Read Dispense dialog opens see the screen sh...

Page 138: ...faults Click OK to return to the Main Menu 11 Select Protocol Save As and give the protocol an identifying name that includes the dispenser number and the dispense volume such as Dispenser 1_80 ul Keep the protocol open 12 Select Protocol Reading again and return to the Read Dispense dialog 13 Double click on the DISPENSE item as shown below to open the Dispense dialog and change the Dispenser to ...

Page 139: ...ngth 405 nm 630 nm Second or Ref Wavelength 750 nm 750 nm Note KC4 automatically creates a multi plate transformation to subtract the second wavelength from the first The name of the resulting data set is Delta OD 5 Select the appropriate Plate Type or set it to 96 WELL PLATE 6 Set First Well and Last Well as follows 80 μl Read 20 5 μl Read First Last Well A1 H4 A5 H12 7 If the reader supports sha...

Page 140: ...nt Qualification BioTek Instruments Inc Sample KC4 Reading Parameters Dialog for the 80 µL Read Protocol using the Synergy HTTR w Injectors Sample KC4 Reading Parameters Dialog for the 20 5 µL Read Protocol using the ELx808 ...

Page 141: ...Dispense Module Tests 119 Synergy HT Operator s Manual Sample KC4 Plate Layout for the 80 µL Read Protocol Sample KC4 Plate Layout for the 20 5 µL Read Protocol ...

Page 142: ...120 Chapter 4 Instrument Qualification BioTek Instruments Inc ...

Page 143: ...weight g 20 µL weight g 5 µL weight g Expected weight 2 5600 g Expected weight 0 6400 g Expected weight 0 1600 g Accuracy Error Accuracy Error Accuracy Error Must be 2 0 P F Must be 5 0 P F Must be 20 0 P F Standard Deviation Standard Deviation Standard Deviation Mean Mean Mean CV CV CV Must be 2 0 P F Must be 7 0 P F Must be 10 0 P F Reader Model Reader S N Tested By Reviewed Approved By Reading ...

Page 144: ...122 Chapter 4 Instrument Qualification BioTek Instruments Inc ...

Page 145: ...d Surfaces 127 Inspecting Cleaning Excitation and Emission Filters 128 Flushing Purging the Fluid Path 129 Running a Dispense Protocol Optional 130 Emptying Cleaning the Tip Priming Trough 132 Cleaning the Priming Plate 132 Cleaning the Internal Components 133 Required Materials 134 Removing the Reader s Shroud 135 Removing the Internal Tubes and Injector Heads 137 Cleaning the Internal Tubes and ...

Page 146: ...of each day ensures optimal performance of the dispense system Perform a visual inspection of the dispensing accuracy before conducting an assay that requires dispense to verify instrument performance It is important to keep the dispensing lines scrupulously clean at all times Take special care when using molecules active at very low concentrations e g enzymes inhibitors Remove any residual reagen...

Page 147: ...r storage Frequency Tasks for Synergy HT Models with Injectors Task Page Daily Quarterly As Needed Clean Exposed Surfaces 127 9 Inspect Clean Excitation and Emission Filters 128 9 Flush Purge the Fluid Path 129 9 Optional Run Dispense Protocol 130 9 Empty Clean Tip Prime Trough 132 9 Clean Priming Plate 132 9 Clean Internal Dispense Tubing and Injector Heads 140 9 Clean Optical Probes 141 9 Clean ...

Page 148: ...t spray it with liquid or use a wet cloth on it Do not allow water or other cleaning solution to run into the interior of the instrument If this happens contact BioTek s Technical Assistance Center L Important Do not apply lubricants to the microplate carrier or carrier track Lubrication on the carrier mechanism or components in the carrier compartment will attract dust and other particles which m...

Page 149: ...s 1 Turn off and unplug the instrument 2 Moisten a clean cotton cloth with water or with water and mild detergent Do not soak the cloth 3 Wipe the plate carrier and all exposed surfaces of the instrument 4 Wipe all exposed surfaces of the dispense module if used 5 If detergent was used wipe all surfaces with a cloth moistened with water 6 Use a clean dry cloth to dry all wet surfaces 7 Reassemble ...

Page 150: ...ecures the emission EM filter wheel Remove each thumbscrew and slide the filter wheel s supporting metal bracket straight out of the compartment Chapter 3 Getting Started contains illustrations for identifying the filter wheels and their unique characteristics This chapter also contains instructions for replacing filters if necessary 3 Inspect the glass filters for speckled surfaces or a halo effe...

Page 151: ...ergy Com KC4 Select System Reader Control 4 Click the Dispenser tab and select Dispenser 1 5 Set the Volume to 5000 L Keep the default prime Rate 6 Click Prime to start the process When the process is complete carefully remove the priming plate from the carrier and empty it 7 Repeat the process for Dispenser 2 Leave the water in the system overnight or until the instrument will be used again Purge...

Page 152: ... the Dispense Volume to 100 µL or an amount to match your assay protocol Select a Rate adjust the rate to support the dispensing volume Click OK to close the dialog and add the Dispense step to the list 3 Add another Dispense step with the same parameters selecting Dispenser 2 4 Add a quick Read step with the following parameters Gen5 requires that a Read step follow the Dispense step Define a par...

Page 153: ...ton 5 In the Read Dispense dialog set Read Mode to Plate 6 Click the Dispense button In the Dispense Step dialog Select Dispenser 1 Set Priming to Before the process and Volume to 5 L Leave Start Time set to 00 00 00 Set the Volume to 100 µL or an amount to match your assay protocol Select a Rate adjust the rate to support the dispensing volume Click OK to close the dialog and add the Dispense ste...

Page 154: ...he trough thoroughly and allow it to dry completely 4 Replace the trough in the microplate carrier When starting a Gen5 Experiment that includes dispensing Gen5 will prompt you to empty the tip prime trough Follow the instructions provided When starting a KC4 Read Dispense protocol KC4 may prompt you to empty the tip priming trough In this case KC4 will automatically open the System Reader Control...

Page 155: ...ce beneath the microplate carrier should be cleaned The procedures in this section should be performed in succession Start with Removing the Reader s Shroud and execute the procedures that meet your needs in the order in which they are presented Finish with Reassembling the Components We recommend running a System Test via Gen5 or KC4 before and after performing these cleaning procedures This will...

Page 156: ... the internal components Phillips head screwdriver 1 8 Allen wrench 3 32 Allen wrench For cleaning the internal dispense tubes and injector heads as well as for wiping the surface under the plate carrier Mild detergent Clean lint free cotton cloths Deionized or distilled water Stylus stored in a plastic cylinder affixed to the rear of the dispense module or reader PN 2872304 For cleaning the optic...

Page 157: ...e PC and the dispense module 1 Disconnect power and all cables Set the external dispense module aside 2 Place the Synergy HT on a work surface that allows you to easily access all sides of the instrument 3 Remove four black Phillips head screws one at the bottom rear corner on each side and two at the top center of the rear panel Note When reinstalling the shroud press down firmly on the top to ma...

Page 158: ...apter 5 Preventive Maintenance BioTek Instruments Inc 4 Stand facing the front of the instrument Grasp both sides of the shroud slide it toward you and pull it straight off the instrument Set the shroud aside ...

Page 159: ...e internal dispense tubes be sure to align the tubing ports with the injector heads as shown in this diagram Look for the SYRINGE 1 and SYRINGE 2 labels on the instrument s rear panel SYRINGE 1 SYRINGE 2 Injector heads 2 white for dispensing fluid into microwells Internal tubes 2 black for delivering fluid to the injector heads Tubing ports 2 for connecting the two internal dispense tubes to the t...

Page 160: ...and injector heads 1 Open the cable clamp to release the tubes 2 Locate the tubing ports on the reader s rear wall Turn each tube s thumbscrew counterclockwise and gently pull the tube from the port 3 Locate the injector heads Turn each tube s thumbscrew counterclockwise to disconnect the tube from the injector head Cable clamp ...

Page 161: ...l Components 139 Synergy HT Operator s Manual 4 Turn the injector heads counterclockwise and gently pull them out of their sockets Note Be sure to seat the injector tips securely when reinstalling See the photo on page 150 ...

Page 162: ...apy water to soften and dissolve any hardened particles 2 Flush each tube by holding it vertically under a stream of water from a faucet To clean the injector heads Do not remove the o ring from the injector head see photos below 1 Gently insert the stylus PN 2872304 into each injector head pipe to clear any blockages The stylus should be stored in a plastic cylinder affixed to the rear of the dis...

Page 163: ...ue to the Noise in the chamber from any spilled fluorescing compounds To access the optical probes the first step is to unplug the reader and remove its shroud cover If you haven t already done this turn to page 135 now for instructions Note For models without injectors the internal chamber and probes are not customer accessible Contact BioTek s Technical Assistance Center with any questions about...

Page 164: ... 38 Internal components to be removed adjusted for cleaning the optic probes Heater thermistor connectors 2 Top optical probe Thumbscrew to access ground wire Incubator housing Screw for lowering and raising the top optical probe Thumbscrews 2 black for holding the incubator housing in place Top probe hanger with two shoulder screws ...

Page 165: ...e page 140 follow these instructions to remove a few more components and then clean the optical probes 1 Disconnect the heater and thermistor wires To do this depress the small tab pictured below and separate the connectors 2 Remove the thumbscrew located in the left rear of the instrument and set it aside This exposes the ground wire Depress tab to separate the connectors ...

Page 166: ...5 Preventive Maintenance BioTek Instruments Inc 3 Lift the ground wire and move it off to the side 4 Locate the two black thumbscrews that hold the incubator housing in place Remove both of them and set them aside ...

Page 167: ...all the way to the bottom Rotate the screw not the ring around it 6 Gently lift the left side of the incubator housing and carefully slide it out Rotate counterclockwise Note When replacing the incubator housing the two forks on its right side should wrap around the holding screws The forks should not slide under the fixed foam housing ...

Page 168: ... Allen wrench to remove the top optical probe s holding screw 8 Gently pull the optical probe up and out of its socket to expose it for cleaning Soak the probe in alcohol for one minute maximum Wipe with lens cleaning tissue and set aside Clean with alcohol and lens tissue ...

Page 169: ...and set them aside 10 Drop the top probe hanger down and slide to the left to remove it Turn the hanger upside down to clean the absorbance lens see instructions on the next page Do not touch the lens with your fingers Inspect the block for spills or other contamination Carefully clean with mild detergent if necessary Shoulder screws Absorbance Lens Probe hanger bottom view ...

Page 170: ... pressure can push the lens out of its holder reinstallation must be performed by BioTek service personnel If the lens does fall out contact BioTek TAC 11 Use a cotton swab moistened with alcohol to gently clean the lens on the top probe hanger 12 Slide the microplate carrier out of the way Use a cotton swab moistened with alcohol to clean the lens on the instrument surface ...

Page 171: ...o at a minimum disconnect the incubator wires detach the ground wire lower the top optic probe hanger and remove the incubator housing steps 1 through 6 2 Manually slide the microplate carrier to the left to engage the support pin and then away from the center surface 3 Moisten do not soak a clean cotton cloth with alcohol water or with water and mild detergent Wipe all sides of the plate carrier ...

Page 172: ... further instructions and photos demonstrating the steps 1 Slide the microplate carrier back into the instrument p 149 2 Insert the two injector heads into their sockets in the top probe hanger Do not touch the absorbance lens with your fingers Ensure that the injector heads are properly seated in the hanger The knurled plastic should sit flush against the hanger surface as shown below 3 Attach th...

Page 173: ...thermistor wires p 143 Be sure to connect wires of the same color 9 Attach the two internal dispense tubes to the tubing ports taking care to align the correct port with the correct injector head p 138 10 Slide the two internal dispense tubes into the cable clamp and close the clamp p 138 11 Review the steps you just performed to make sure the components have been properly reassembled 12 Slide the...

Page 174: ...152 Chapter 5 Preventive Maintenance BioTek Instruments Inc ...

Page 175: ...rials 155 Procedure for Models without Injectors 156 Routine Procedure for Models With Injectors 157 Clean Exposed Surfaces 157 Decontaminate the Fluid Lines 158 Rinse the Fluid Lines 159 Clean the Internal Tubing and Injector Heads 159 Clean the Tip Priming Trough and Priming Plate 160 Alternate Procedure for Models with Injectors 161 ...

Page 176: ...endations of the Centers for Disease Control and Prevention CDC Neither BioTek nor the CDC assumes any liability for the adequacy of these solutions and methods Each laboratory must ensure that decontamination procedures are adequate for the Biohazard s they handle Wear prophylactic gloves when handling contaminated instruments Gloved hands should be considered contaminated at all times keep glove...

Page 177: ...lcohol as an alternative to bleach Deionized or distilled water Safety glasses Surgical mask Protective gloves Lab coat Biohazard trash bags 125 ml beakers Clean lint free cotton cloths Additional materials for models with injectors Phillips head screwdriver Small brush for cleaning the tip priming trough and priming plate Optional Mild detergent ...

Page 178: ...ach are a concern Be sure to check the percent NaClO of the bleach you are using this information is printed on the side of the bottle Commercial bleach is typically 10 0 NaClO if this is the case prepare a 1 20 dilution Household bleach is typically 5 0 NaClO if this is the case prepare a 1 10 dilution 3 Moisten a cloth with the bleach solution or alcohol Do not soak the cloth 4 Manually open the...

Page 179: ... Clean Exposed Surfaces 1 Turn off and unplug the instrument 2 Prepare an aqueous solution of 0 50 sodium hypochlorite bleach As an alternative 70 isopropyl alcohol may be used if the effects of bleach are a concern Be sure to check the percent NaClO of the bleach you are using this information is printed on the side of the bottle Commercial bleach is typically 10 0 NaClO if this is the case prepa...

Page 180: ...se module s outlet tubes from the instrument s rear panel Place the ends of the outlet tubes in an empty beaker and set the beaker on the work surface 4 Launch Gen5 or KC4 select System Reader Control and click the Dispenser tab 5 Select Dispenser 1 enter a Volume of 5000 µl and keep the default dispense Rate 6 Place the priming plate on the carrier it is not used but the reader requires its prese...

Page 181: ...e for 10 minutes and then run one prime cycle with 5000 µl This delay will allow any residual sodium hypochlorite to diffuse into the solution and be flushed out with the next prime 7 Empty the beaker containing the outlet tubes 8 Wipe all surfaces with deionized water 9 Discard the used gloves and cloths using a Biohazard trash bag and an approved Biohazard container Clean the Internal Tubing and...

Page 182: ...an the corners of the trough and plate 3 To decontaminate soak the trough and plate in a container of 0 5 sodium hypochlorite or 70 isopropyl alcohol for 20 to 30 minutes 4 If decontaminating in bleach solution remove the trough and plate and thoroughly rinse with DI water If decontaminating with alcohol remove the trough and plate and let them air dry 5 Discard the used gloves and cloths using a ...

Page 183: ...bes and Injector Heads 2 Prepare an aqueous solution of 0 50 sodium hypochlorite bleach As an alternative 70 isopropyl alcohol may be used if the effects of bleach are a concern Be sure to check the percent NaClO of the bleach you are using this information is printed on the side of the bottle Commercial bleach is typically 10 0 NaClO if this is the case prepare a 1 20 dilution Household bleach is...

Page 184: ... the bottom of the syringe drive Gently remove the syringe and store it in its original packaging see Chapter 2 Installation 7 Moisten a cloth with DI or distilled water and wipe all surfaces that have been cleaned with the bleach solution or alcohol 8 Rinse all tubing and the syringes with DI water 9 Use a clean dry cloth to dry all wet surfaces on the instrument and the Dispense module 10 Reasse...

Page 185: ...eypad or its own user interface BioTek s Gen5 and KC4 software packages support all Synergy HT models and are used for both instrument control and data reduction For information on serial protocol commands and or system integration please contact your BioTek sales representative or BioTek Instruments Inc ...

Page 186: ...164 Appendix B Computer Control BioTek Instruments Inc ...

Page 187: ...PC Error Codes 166 Fatal Errors 167 Non Fatal Errors 168 Home Sensor Initial Find Errors 0100 0307 168 Home Sensor Verification Errors 0400 0409 172 Saturation Errors 0500 0616 174 Absorbance Reader Noise Errors 0700 0A16 180 Internal Self Test Errors 0D00 2918 184 Other Errors 2A00 4000 196 Status String Format 202 ...

Page 188: ...tes a more serious error with the memory or processing In this case the instrument may be unable to respond turn the instrument off and on again You may be able to use the instrument after restarting it See Fatal Errors on the next page If an error code is displayed run a System Test for diagnostic purposes If you re using Gen5 select System Diagnostics Run System Test If you re using KC4 select S...

Page 189: ...n addition to being used by the motors is also used in microsecond timing for time resolved measurements A400 Failed code checksum test on power up A500 DR steps alloc free error assay number This error indicates old basecode recommend upgrading A502 24V power dropped below safe level A600 Data flash write timed out A700 Data flash read back did not match write A800 Code flash write timed out A900...

Page 190: ... Possible Causes User aborted read from Gen5 or KC4 User aborted from another serial interface 0200 X axis motor did not find the home opto sensor transition This error indicates that a motor was not able to move to its home position as registered by feedback from an optical sensor Note This error does not look for the mid or XY sensor See 2A01 for mid sensor See 2400 for XY sensor Probable Causes...

Page 191: ...id not home Probable Causes Filter wheel is not inserted into to the EX assembly Filter wheel is not moving due to an obstruction or because the filter is not clipped in Filter wheel is not moving because gear teeth on the filter wheel are binding with the gear teeth on the motor Defective or broken Hall Effect sensor Defective Motor Controller PCB or cable Motors failed due to the bearings 0203 E...

Page 192: ...e changer assembly is not able to move because a foreign object is preventing the movement 0208 Syringe 0 did not home also known as the first syringe drive Probable Causes Linear way is dirty or lack of lubrication is causing the bearings to jam The lead screw is loose on the motor shaft because the glue is no longer bonding it to the motor shaft Defective optical sensor Defective motor Motor Con...

Page 193: ...ax light to no light was never found Probable Causes Absorbance mode The rail nylon bushings or bearings are dirty causing the carrier to jam The Autocal jig is not in the carrier Order sorting filter wheel is jammed and is not able to turn to the open hole Fluorescence mode The rail nylon bushings or bearings are dirty The Autocal jig is not in the carrier or mirrors are facing the wrong directio...

Page 194: ...roperly The Z axis lead screw is no longer glued to the motor shaft The upper fiber bundle is hitting the top of the shroud due to not being properly tied down 0307 Probe changer motor did not find home This error code indicates old basecode Recommend upgrading Probable Causes An obstruction is preventing the motor from moving The motor and or motor drive circuit is defective The sensor and or sen...

Page 195: ... is loose Filter wheel is binding against the motor gear Motors failed due to the bearings 0405 Monochromator failed to find the zero order position White light or positional verify The order sorting filter wheel is homed and moved to the open hole position The monochromator is moved until the optical system detects saturation It is then moved to the point where there is no saturation and then mov...

Page 196: ... failed For certain older dispensers opto cable 7330506 may need to be replaced with 7120734 The linear way and bearings are dirty or lack sufficient grease The lead screw is loose on the motor shaft because the glue is no longer bonding it to the motor shaft The assembly is not able to move because a foreign object is preventing the movement Saturation Errors 0500 0616 These errors occur in sever...

Page 197: ...o the lamp power supply Scenario 4 fluorescence autocalibration The surface of the autocalibration jig is too reflective Return the jig to BioTek for a replacement jig The carrier did not move far enough to block the light beam Troubleshoot the carrier movement 0501 Light beam saturated light Air measurement channel reading reached 65535 or PMT Relative Fluorescing Units RFU reached FFFF 99999 Thi...

Page 198: ...The PMT is defective The connector from the PMT base to the analog board was not built correctly The ground shield is not grounding properly The PMT base is defective The PMT analog PCB is defective Scenario 2 Reference channel The monochromator mirror grating is damaged Absorbance analog PCB has intermittently failed Order sorting filter wheel is jammed not aligning the correct bandpass filter wi...

Page 199: ... home Flash bulb skipped a flash due to a defective lamp connection or power supply Order sorting filter wheel is jammed not aligning the through hole with the light path Monochromator is defective Absorbance analog PCB has intermittently failed Scenario 2 Reference channel The monochromator mirror grating is damaged Absorbance analog PCB has intermittently failed Missed flashes or an erratic flas...

Page 200: ...ntermittently failed Missed flashes or an erratic flash lamp Order sorting filter wheel is jammed not aligning the correct bandpass filter with the light path or the through hole is allowing white light to pass Voltage to the lamp has increased due to failure of the lamp power supply or the motor power supply PCB sent the wrong voltage request to the lamp power supply Scenario 2 Spectral scan read...

Page 201: ...ail if Reference signal 40000 and Gain 1 Note The order of the last two digits is Filter Channel This is not consistent with other error codes This error indicates that the absorbance reference channel gain for a specific wavelength during Time Resolved readings is out of the range necessary to ensure the lambda performance to specifications The second to last number is the lambda table position n...

Page 202: ...etrating the measurement chamber The bottom and top shrouds are part of the electrical shielding Verify that the shrouds are installed and properly fastened The coaxial cable ground between the reference channel and the absorbance analog PCB may be floating or disconnected There may be an ambient light leak Ensure the plate carrier door and the front hinged door are properly closed Analog PCB fail...

Page 203: ...ng the system test the background electronic signal that was detected is outside of acceptable limits at maximum gain when blocking the light Probable Causes If noise Max is 20000 The photodetector is too noisy and is defective Absorbance channel analog PCB is defective A faulty analog PCB or faulty internal grounding may cause internal electronic noise There may be an ambient light leak Ensure th...

Page 204: ...nnel photodetector is defective A faulty analog PCB or faulty internal grounding may cause internal electronic noise There may be an ambient light leak Ensure the plate carrier door and the front hinged door are properly closed Electrical noise may be penetrating the measurement chamber The bottom and top shrouds are part of the electrical shielding Verify that the shrouds are installed and proper...

Page 205: ...nd analog PCB is defective or disconnected Measurement channel photodetector is defective If failed 20000 Measurement channel photodetector is defective A faulty analog PCB or faulty internal grounding may cause internal electronic noise There may be an ambient light leak Make sure the plate carrier door and the front hinged door are properly closed Electrical noise may be penetrating the measurem...

Page 206: ...t below This error indicates the Air reading at the time of the plate read was 50 of the Air reading at the time of the optic test The last number is the lambda table position number Probable Causes Flash lamp has missed flashes or there is an erratic flash during the blank read Dirty optics or spilled substance on the optics Order sorting filter wheel is jammed not aligning the correct bandpass f...

Page 207: ... or 2 0 Probable Causes Lamp alignment or lamp power supply failure Defective reference channel analog PCB or absorbance channel analog PCB or the cable in between Order sorting filter wheel is jammed not aligning the correct bandpass filter with the light path and white light is passing through or the filter is degraded and is not passing enough light energy or is blocking the light Damaged optic...

Page 208: ...th the light path and white light is passing through or the filter is degraded and is not passing enough light energy or is blocking the light Damaged optic spray 0F11 0F16 Measurement channel correction outside limits See criteria in text below See Delta Air Dark on self test This error can indicate one of the following error scenarios During a system test the Delta was 8000 for one of the wavele...

Page 209: ...top probe absorbance Probable Causes The 7080400 PCB was changed and the Flash memory does not have the calibration values loaded Performing the autocalibration procedure will correct this 130x Motor not homed successfully This error indicates that the motor is not at home At the beginning of the motor_position function the basecode checks to see if the motor is homed If it is not homed an error i...

Page 210: ... zone 1 1 2 2 3 4 4 8 see table below for more information Error code Zone 1501 Zone 1 1502 Zone 2 1503 Zone 1 and 2 1504 Zone 3 1505 Zone 1 and 3 1506 Zone 2 and 3 1507 Zone 1 2 and 3 1508 Zone 4 1509 Zone 1 and 4 150A Zone 2 and 4 150B Zone 1 2 and 4 150C Zone 3 and 4 150D Zone 1 3 and 4 150E Zone 2 3 and 4 150F Zone 1 2 3 and 4 Probable Causes Thermistor is defective for that zone Heating pad i...

Page 211: ...1 1 2 2 3 4 4 8 see table below for more information Error code Zone 1511 Zone 1 1512 Zone 2 1513 Zone 1 and 2 1514 Zone 3 1515 Zone 1 and 3 1516 Zone 2 and 3 1517 Zone 1 2 and 3 1518 Zone 4 1519 Zone 1 and 4 151A Zone 2 and 4 151B Zone 1 2 and 4 151C Zone 3 and 4 151D Zone 1 3 and 4 151E Zone 2 3 and 4 151F Zone 1 2 3 and 4 Probable Causes Thermistor is defective for that zone Motor power supply ...

Page 212: ...netic interval not correct for selected options This error indicates one of the following scenarios Kinetic interval in the current assay is too short Kinetic interval for a fluorescence plate read is impossible for the given parameters Kinetic interval for an absorbance read the total time or kinetic interval 0 Kinetic interval for a fluorescence kinetic interval is too big 99999 to transmit Prob...

Page 213: ...citation and emission filter wheels designated by the protocol Equation used for overlapping If EM wavelength is EX if EX wavelength bandpass EM wavelength bandpass then fail else if EX wavelength is EM if EM wavelength bandpass EX wavelength bandpass then fail Note Only 2 filter sets can be sent when developing software not Gen5 or KC4 1F01 is for the first set 1F02 is for the second set 2100 Inv...

Page 214: ...serviced soon Probable Causes X axis rail is dirty The nylon slider bushings are worn and causing too much friction or dirt in the roller bearings is causing the bearings to jam Y axis rail is dirty The bearings are dirty and worn causing too much friction Defective or broken optical sensor Defective Motor Controller PCB Bent opto flag on carrier Carrier feet have been adjusted to the point where ...

Page 215: ...nging the width to 84320 will alleviate the error message and will not affect the results In KC4 v3 4 r16 the width was changed to 84320 and the 2600 error does not appear when running a protocol that specifies the Corning Costar 12 Well plate type 280x Motor currently in use This error indicates that the motor is not available for this model or already has a task assigned to it At the beginning o...

Page 216: ...age Reference Mtr This tests the voltage across a sense resistor in series of the 40 volts it is monitoring the current This test is performed during self test Probable Causes The motor power supply PCB is defective The sense resistor is damaged on the motor power supply PCB 2904 3 5V Flash Power Supply reference voltage is out of range 1399 1453 See self test Voltage Reference Min This tests the ...

Page 217: ...istor in series of the 8 00 volts it is monitoring the current This test is performed during self test Probable Causes The motor power supply PCB is defective The sense resistor is damaged on the motor power supply PCB 2911 2918 VRef instability This error indicates that one of the channels voltage reference feeding the A D converter is unstable i e A D max A D min 8 after 8 attempts See the table...

Page 218: ...air bubble the syringe must be in the opto sensor Note This error can apply to either Syringe 1 or Syringe 2 Probable Causes Linear way is dirty or needs lubrication Defective or broken optical sensor Syringe was not installed correctly or was not cleaned causing stress on syringe movement by not allowing the syringe to move to the home position The glue between the lead screw and the motor has se...

Page 219: ...nge to move freely The glue between the lead screw and the motor has separated 2B05 Number of microsteps requested for move too large This error indicates that during a dispense or move home function the number of steps to move was the maximum syringe travel 944 full steps Note This error can apply to either Syringe 1 or Syringe 2 Probable Causes Program or user error 2B0A Trough plate not in carr...

Page 220: ...ge needs to be calibrated Note This error can apply to either Syringe 1 or Syringe 2 Probable Causes Factory default data is set the syringe needs to be calibrated before use or calibration values need to be entered from the syringe drive Values entered are corrupted or incorrect 2D00 During a dispense read the reader missed the start of the well read This error indicates that during a dispense re...

Page 221: ... spacing not valid for dispensing Well spacing in any direction is 0 320 or 8 13 mm Probable Causes A 384 well plate was selected for a Dispense protocol 2D0E During a dispense read the reader missed the start of the plate read This error indicates that during a dispense read the reader was not able to start the read Probable Causes The processor was busy running other tasks A previous task took l...

Page 222: ...the Excitation filter slot when a non TR fluorescence dispense read is selected Probable Causes The Time Resolved cartridge is not installed in the EX position The Hall Effect sensor is defective Instrument does not have the TR functionality and the DIP switch setting 4 on the 7080400 PCB is open and the DIP switch setting 1 is open on the 7090410 PCB 3100 Plate read took longer than kinetic inter...

Page 223: ...B 0C 0D 0E 0F 10 11 12 13 14 15 16 17 18 B 19 1A 1B 1C 1D 1E 1F 20 21 22 23 24 25 26 27 28 29 2A 2B 2C 2D 2E 2F 30 C 31 32 33 34 35 36 37 38 39 3A 3B 3C 3D 3E 3F 40 41 42 43 44 45 46 47 48 D 49 4A 4B 4C 4D 4E 4F 50 51 52 53 54 55 56 57 58 59 5A 5B 5C 5D 5E 5F 60 E 61 62 63 64 65 66 67 68 69 6A 6B 6C 6D 6E 6F 70 71 72 73 74 75 76 77 78 F 79 7A 7B 7C 7D 7E 7F 80 81 82 83 84 85 86 87 88 89 8A 8B 8C 8...

Page 224: ...ailed positional verify SATURATION ERR 0500 A D signal saturated filter channel FILTER GAIN ERR 0600 filter gain out of range NOISE TEST ERR 0700 reader channel failed noise test OFFSET TEST ERR 0800 reader channel failed offset test DARK RANGE ERR 0900 read time channel filter dark out of range AIR RANGE ERR 0A00 read time channel filter air blank out WAVECAL DATA ERR 0D00 wavelength calibration ...

Page 225: ... D ready transition DOOR LOCK ERR 3300 motor required carrier outside when locked in PMT WELL ERR 4000 overload at well A01 P24 Test Type Codes lowest digit in returned error code FAIL VTEST HI 1 higher PMT voltage level incorrect FAIL VTEST LO 2 lower PMT voltage level incorrect FAIL WELL TEST 5 PMT saturation at well FAIL BKGRND TEST 8 PMT saturation during background overload test Vref Test Cha...

Page 226: ...t should be off SENSOR CLEAR COUNT 3 sensor clear count out of range FMEA TEST FAIL 4 clear count measured during aspirate operation deviated more than allowed range from initial value MOVE OVERRANGE 5 number of microsteps requested for move too large SYR NOT HOME 6 syringe logical position not at home INVALID POSITION 7 invalid position passed to position syringe function INVALID OPERATION 8 inva...

Page 227: ...issed its scheduled start time INVALID TIP PRIME 0F tip prime conflicts with tip prime rules NUM READ EVENTS 10 number of read events defined invalid READ START TIME 11 read event start time out of range NUM SAMPLES 12 number of samples out of range SAMPLING INTERVAL 13 sampling interval out of range NUM PLATE READS 14 number of plate mode kinetic reads out of range PLATE MODE READ TIME 15 plate m...

Page 228: ...column of the offending well can be extracted from the well code by applying the following formula examples use 8 x 12 96 well plate 1 Convert the ASCII hex string to a decimal equivalent Ex 057 indicates 57 hex yielding a well code of 87 decimal 2 Row well code columns in plate rounded up to a whole number Ex 87 12 7 25 indicating row 8 or H 3 Column well code row 1 columns in plate Ex 87 8 1 12 ...

Page 229: ...ions This appendix contains BioTek s published specifications for the Synergy HT Specifications 208 Microplates 208 Hardware and Environmental 208 Absorbance 209 Fluorescence 211 Incubation 213 Shake 213 Injector Model 213 ...

Page 230: ...y require an adapter All models read 6 12 24 48 96 and 384 well microplates Injector models dispense to 6 12 24 48 and 96 well microplates Hardware and Environmental Light Source Absorbance Xe flash light source 10 W max average power Life 1 billion flashes Fluorescence Tungsten quartz halogen 20 W Life 1000 hours Dimensions 16 D x 15 W x 10 H 40 6 cm x 38 cm x 25 4 cm Weight 38 lb 17 kg Environme...

Page 231: ...e 96 and 384 well plates Linearity 0 000 to 2 000 OD 1 0 Normal and Rapid modes 96 well plates 0 000 to 2 000 OD 2 0 Normal and Rapid modes 384 well plates 2 000 to 2 500 OD 3 0 Normal and Rapid modes 96 and 384 well plates 2 500 to 3 000 OD 3 0 Normal mode 96 well plates 0 000 to 1 000 OD 1 0 Sweep mode 96 and 384 well plates Repeatability 0 000 to 2 000 OD 1 0 0 005 OD Normal and Rapid modes 96 ...

Page 232: ...ch row past the optics channel without stopping collecting data with a single flash at each well as it goes by Read Timing The following read times are based on a single or dual wavelength measurement Actual reading times can vary depending upon the wavelength read mode and other reading parameters selected In Normal read mode see above the optical density of the solution affects timing as well En...

Page 233: ...ptical Probes The Synergy HT is configured with a variety of probe sizes 1 5 and 3 mm probes can be installed in either the top or bottom positions the 5 mm probe can only be installed in the bottom position Sensitivity 5 mm probe Sodium Fluorescein SF Bottom reading 10 pg ml solution of Sodium Fluorescein in PBS 150 µL per well signal to noise ratio greater than 2 0 Excitation 485 20 Emission 530...

Page 234: ...tation 360 40 Emission 460 40 Corning Costar black strips 1 5 mm probe Sodium Fluorescein SF Bottom reading 40 pg ml solution of Sodium Fluorescein in PBS 150 µL per well signal to noise ratio greater than 2 0 Excitation 485 20 Emission 530 25 Hellma 96 well quartz plate Propidium Iodide PI Bottom reading 250 ng ml solution of Propidium Iodide in PBS 50 µL per well signal to noise ratio greater th...

Page 235: ... specifications apply to Synergy HT models with injectors Plate Type Dispenses to standard 6 12 24 48 and 96 well microplates with 128 x 86 mm geometry Dispense Volume Range 5 1000 µL with a 5 20 µL tip prime Accuracy Dispensing deionized water with 0 1 Tween 20 at room temperature 1 µL at 5 50 µL 2 at 51 1000 µL Precision Dispensing a 200 µL solution of deionized water 0 1 Tween 20 and dye at roo...

Page 236: ...214 Appendix D Specifications BioTek Instruments Inc ...

Page 237: ...gure 40 shows the location of the microplate carrier in reference to the exterior surfaces of the Synergy HT and the mounting holes on the bottom The illustration should facilitate system setup with a robotic instrument such as the BioStack Microplate Stacker ...

Page 238: ...s for Robotic Interface If you have purchased the BioStack to operate with the Synergy HT special alignment hardware is included in the BioStack alignment kit for correct positioning of the Synergy HT with the BioStack Refer to the Installation chapter in the BioStack Operator s Manual for instructions ...

Page 239: ... 000 17 132 000 3 490 3 400 5 070 A1 000 500 5 626 000 362 10 612 000 555 15 345 Figure 40 Instrument dimensions mounting holes and carrier position TOP VIEW SIDE VIEW BOTTOM VIEW The two arrows point to special mounting holes for alignment caps for operation with the BioStack ...

Page 240: ...218 Appendix E Instrument Dimensions for Robotic Interface BioTek Instruments Inc ...

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