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102

| Chapter

Instrument Qualification Procedures

Pipette 150 μL of buffer solution into wells C2–F5. Discard the tips.

Pipette 150 μL of the

1 nM SF

solution into column 1.

Pipette 150 μL of the 1 nM SF solution into column 2. Mix the wells using the
pipette. Do not discard the tips.

Aspirate 150 μL from column 2, and dispense into column 3. Mix the wells using
the pipette. Do not discard the tips.

Aspirate 150 μL from column 3, and dispense into column 4. Mix the wells using
the pipette. Do not discard the tips.

Aspirate 150 μL from column 4, and dispense into column 5. Mix the wells using
the pipette. Do not discard the tips.

Aspirate 150 μL from column 5, and discard the tips.

Alternate/Supplemental Tests Using Methylumbelliferone (MUB)

As an alternative to using Sodium Fluorescein, Methylumbelliferone (“MUB”) can be used
to test the fluorescence system.

Required Materials

Manufacturer part numbers are subject to change over time.

Synergy LX User Manual
1501000N Revision A

BioTek Instruments, Inc.

Summary of Contents for Synergy LX

Page 1: ...gy LX Multi Mode Microplate Reader User Manual ERRATA NOTICE This document contains references to BioTek Please note that BioTek is now Agilent For more information go to www agilent com lifesciences biotek ...

Page 2: ... Manual ERRATA NOTICE This document contains references to BioTek Please note that BioTek is now Agilent For more information go to www agilent com lifesciences biotek Document part number 1501000N revision A BioTek Instruments Inc December 2021 ...

Page 3: ... Technical Support 6 Installation 7 Important Information 8 1 Unpack the Box and See What s Inside 9 2 Remove the Shipping Screw and Insert the Light Blocking Plug 10 3 Gen5 control only Prepare the Host Computer 12 4 Gen5 control only Connect the Host Computer and Reader 13 5 Install the Power Supply 13 6 Turn on the Reader and Run the Power Up System Test 15 7 Gen5 control only Start Gen5 and Te...

Page 4: ...reen 55 Inspect Clean Filter Cubes 56 Clean the Air Filter 57 Replace the Halogen Bulb Assembly 59 Decontamination 60 Instrument Qualification Process 62 System Test 63 Plate Shaker Test 63 Absorbance Testing 64 Fluorescence Testing 73 Luminescence Test 81 Instrument Qualification Procedures 85 Overview 86 IQ OQ PQ Description 86 Recommended Qualification Schedule 88 System Test 89 Absorbance Plat...

Page 5: ...12 Luminescence Specifications 112 Sample Reports 113 Sample System Report 114 Sample Absorbance Test Plate Report 116 Sample Assay Results 117 Error Conditions 119 Overview 120 Error Codes 120 Safety Information 146 Safety Notices 147 In This Book 174 4 Contents Synergy LX User Manual 1501000N Revision A BioTek Instruments Inc ...

Page 6: ...ermission of BioTek Instruments Inc 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 document will be incorporated in new editions of the publication No responsibility is assumed by BioTek for the use or reliability of software or equipment that is not supplied by Bio...

Page 7: ...ice www agilent com en support bio tac agilent com Instrument service and repair is available worldwide at one of our international service centers and in the field at your location UK Responsible Person UKRP Agilent LD UK Ltd 5500 Lakeside Cheadle Royal Business Park Cheadle Cheshire SK8 3GR 6 Contact Information Synergy LX User Manual 1501000N Revision A BioTek Instruments Inc ...

Page 8: ...aboratory samples according to instructions and specific recommendations included in the assay package insert for the test to be conducted Failure to conduct Quality Control checks could result in erroneous test data Warranty and Product Registration Review the warranty information that shipped with your product Register your product to ensure you receive important information updates about the pr...

Page 9: ... not proceed beyond a CAUTION notice until the indicated conditions are fully understood and met Warnings and Precautions Electrical Hazards Internal Voltage Always turn off the power switch and unplug the power supply before cleaning the outer surface of the instrument Power Rating The instrument s power supply or power cord must be connected to a power receptacle that provides voltage and curren...

Page 10: ...touchscreen Environmental Conditions Do not expose the instrument to temperature extremes For proper operation temperature near the instrument should remain within the range in the Specifications section of this document Performance may be adversely affected if temperatures fluctuate above or below this range Sodium Hypochlorite Do not expose any part of the instrument to the recommended diluted s...

Page 11: ... methylene chloride and hydrocarbons These will permanently damage the touchscreen Avoid fibrous materials such as paper towels which can scratch the touchscreen Dirt particles and cleaning agents will get trapped in the scratches Never spray solutions directly on the touchscreen Spare Parts Only approved spare parts should be used for maintenance The use of unapproved spare parts and accessories ...

Page 12: ... and PC based software in conjunction with their specific assay s This evaluation must include the confirmation that performance characteristics for the specific assay s are met Unspecified Use Failure to operate equipment according to the guidelines and safeguards specified in the product user documentation could result in a hazardous condition Use of labware other than described in this document...

Page 13: ...et des précautions consultez le mode d emploi Achtung lesen Sie die Gebrauchsanweisung für wichtige Vorsichtshinweise wie Warnungen und Sicherheitsvorkehrungen Attenzione consultare le istruzioni per l uso per importanti informazioni cautelative come avvertenze e precauzioni Precaución consulte las instrucciones de uso para obtener información importante como advertencias y precauciones Warning Bi...

Page 14: ...hgefahr Avvertenza rischio di pizzicamento Advertencia peligro de atrapamiento Warning Hot surface Waarschuwing heet oppervlak Avertissement surface chaude Warnung heiße Oberfläche Avvertenza superficie molto calda Advertencia superficie caliente Synergy LX User Manual 1501000N Revision A BioTek Instruments Inc ...

Page 15: ... déchets d équipement électrique et électronique DEEE ou aux dispositions locales Entsorgungshinweis Entsorgen Sie das Gerät gemäß der Richtlinie 2012 19 EU für Elektro und Elektronik Altgeräte WEEE bzw den Landesvorschriften Avviso per lo smaltimento smaltire lo strumento in base alla Direttiva 2012 19 EU sui rifiuti di apparecchiature elettriche ed elettroniche WEEE o le ordinanze locali Aviso d...

Page 16: ...spannung Marcatura CE Indica la conformità ai requisiti della Direttiva 2014 30 UE sulla Compatibilità Elettromagnetica e della Direttiva 2014 35 UE sulla Bassa Tensione Marcado CE indica el cumplimiento de los requisitos de la Directiva 2014 30 UE sobre compatibilidad electromagnética y la Directiva 2014 35 UE sobre baja tensión Date of manufacture Productiedatum Date de fabrication Herstellungsd...

Page 17: ...t pas de mutation dans des conditions de fonctionnement normales pendant 40 ans Dieses Produkt entspricht der Umweltschutz Nutzungsdauer gemäß der Definition im Electronic Industry Standard SJ T11364 2006 der Volksrepublik China Giftige oder gefährliche Stoffe werden unter normalen Betriebsbedingungen 40 Jahre lang nicht austreten oder mutieren Questo prodotto è conforme al periodo di utilizzo del...

Page 18: ...es applicables aux produits vendus en Grande Bretagne Die Kennzeichnung UK Conformity Assessed ist ein Zertifizierungszeichen das die Konformität mit den geltenden Anforderungen für in Großbritannien verkaufte Produkte anzeigt Il marchio UKCA conformità valutata del Regno Unito è un marchio di certificazione che indica la conformità ai requisiti applicabili per i prodotti venduti in Gran Bretagna ...

Page 19: ... les procédures d évaluation de la conformité EAC MED ist ein Zertifizierungszeichen zur Kennzeichnung von Produkten die allen Sicherheits und Qualitätsanforderungen der Eurasischen Zollunion entsprechen Das bedeutet dass die EAC MED gekennzeichneten Produkte alle Anforderungen der entsprechenden technischen Bestimmungen erfüllen und alle Konformitätsbewertungsverfahren bestanden haben EAC MED è u...

Page 20: ...formité RCM Exigences en matière d étiquetage ACMA Das Produkt entspricht den australischen Kommunikationsanforderungen EESS Kennzeichnung Regulatory Compliance Mark RCM ACMA Kennzeichnungsanforderungen Il prodotto è conforme ai requisiti Australian Communications Requirements EESS marchio di conformità alle normative Requisiti di etichettatura ACMA El producto cumple con los requisitos de comunic...

Page 21: ...zierungszeichen KC bezeichnet das koreanische Produktkonformitätszeichen für Sicherheit und EMV Funk SAR von elektrischen und elektronischen Geräten Die EMV Anforderungen gelten für Agilent Produkte Il marchio Korea Certification KC indica il marchio di conformità del prodotto Corea per la sicurezza e EMC Radio SAR di apparecchiature elettriche ed elettroniche I requisiti EMC vengono applicati ai ...

Page 22: ...ls 21 Humidity limitation Vochtigheidsbeperking Limitation d humidité Feuchtigkeitsbegrenzung Limitazione dell umidità Limitación de humedad Synergy LX User Manual 1501000N Revision A BioTek Instruments Inc ...

Page 23: ...oratory Use Part 1 General Requirements EN 61010 1 Safety Requirements for Electrical Equipment For Measurement Control and Laboratory Use Part 1 General Requirements EN 61010 2 010 Safety Requirements for Electrical Equipment For Measurement Control and Laboratory Use Part 2 010 Particular requirements for laboratory equipment for the heating of materials CAN CSA C22 2 No 61010 1 Safety Requireme...

Page 24: ...N IEC 61326 1 and 61326 2 6 ELECTRICAL EQUIPMENT FOR MEASUREMENT CONTROL AND LABORATORY USE PART 1 GENERAL REQUIREMENTS FOR NON IVD LISTED PRODUCTS Ingress Protection Code IP 20 Protected against solid foreign objects of 12 5 mm diameter and greater No protection against water Disposal Dispose of the instrument according to Directive 2012 19 EU on waste electrical and electronic equipment WEEE or ...

Page 25: ...hapter 1 Introduction This chapter introduces the Synergy LX and provides contact information for technical assistance Product Description 2 Package Contents 3 Optional Accessories 4 Technical Support 6 ...

Page 26: ...Take3 Trio and 60 72 and 96 well Terasaki plates Gen5 also offers extensive data analysis and reporting and exporting capabilities Part Number Absorbance Fluorescence Luminescence Touchscreen SLXA SN SLXF SN SLXFA SN SLXATS SN SLXFTS SN SLXFATS SN The models with fluorescence capability use a user replaceable halogen bulb as their light source models with absorbance capability use a xenon flash bu...

Page 27: ...by country USB cable 75108 Blank USB flash drive Touchscreen models 1560531 7 64 hex wrench for opening bulb access panel 48169 3 32 hex wrench for removing shipping screw 48570 LUM cube F models 1505003 Optional accessories per the sales order unless shipped separately Synergy LX User Manual 1501000N Revision A BioTek Instruments Inc ...

Page 28: ...0534N Take3 Micro Volume Plate TAKE3 Take3 Trio Micro Volume Plate TAKE3TRIO BioCell Adapter Plate 7270512 BioCell Quartz Vessel 7272051 Terasaki Adapter Plate 7330531 RP D10 Seiko thermal printer power cord for use with touchscreen models 02434 varies according to country of use Halogen replacement lamp kit fluorescence capable models 1500535 Filter cubes filters and mirrors Contact BioTek for pa...

Page 29: ...tinamide Adenine Dinucleotide reduced form 98233 or Sigma N6785 10VL Fluorescence Liquid Tests Test Kits Kit for FI tests using Sodium Fluorescein 7160013 Kit for FI tests using Methylumbelliferone 7160012 Individual Materials Sodium Fluorescein Powder 1 mg vial 98155 Methylumbelliferone 10 mg vial 98156 Carbonate Bicarbonate Buffer CBB capsules Sigma 3041 Phosphate Buffered Saline PBS tablets pH ...

Page 30: ... instruments needing repair the specific steps that produce your problem and any error codes displayed on the touchscreen or in Gen5 see also Appendix B Error Conditions l Gen5 users A text file of the diagnostic history of the instrument select System Diagnostics History then select the appropriate file and click Export Running a system test when a problem occurs provides valuable information for...

Page 31: ...ee What s Inside 9 2 Remove the Shipping Screw and Insert the Light Blocking Plug 10 3 Gen5 control only Prepare the Host Computer 12 4 Gen5 control only Connect the Host Computer and Reader 13 5 Install the Power Supply 13 6 Turn on the Reader and Run the Power Up System Test 15 7 Gen5 control only Start Gen5 and Test Communication 15 Operational Performance Qualification 16 Repackaging and Shipp...

Page 32: ...ng materials Be sure to use packaging materials supplied by the manufacturer when shipping the reader Using other forms of commercially available packaging or failing to follow the repackaging instructions may void your warranty l During the unpacking process inspect the packaging reader and accessories for shipping damage If the reader is damaged notify the carrier and your BioTek representative ...

Page 33: ...l items that you ordered 2 Remove the accessories tray and set it aside Lift the reader out of the box and set it on a stable level surface Remove the plastic bag and remove the desiccant pack 3 Save the box bag foam top tray and corner cubes and accessories tray in case you need to send the reader to BioTek for service Synergy LX User Manual 1501000N Revision A BioTek Instruments Inc ...

Page 34: ...be removed before operating the instrument and reinstalled before repackaging the instrument for shipment 1 From the back of the reader Remove the shipping screw At the front of the reader 2 Open the upper access door Insert the shipping screw into its storage location Synergy LX User Manual 1501000N Revision A BioTek Instruments Inc ...

Page 35: ...ht Blocking Plug 11 3 Below this storage location remove the rubber light blocking plug At the back of the reader Insert the plug into the hole left open during step 1 Twist to seat Synergy LX User Manual 1501000N Revision A BioTek Instruments Inc ...

Page 36: ... installed and configured Please follow the instructions provided with Gen5 to install the software and USB driver Gen5 software versions 3 11 and higher require Windows 10 You must have administrator privileges to install Gen5 Log in to Windows as Administrator or consult your IT department for assistance Synergy LX User Manual 1501000N Revision A BioTek Instruments Inc ...

Page 37: ...Grounding Never use a plug adapter to connect primary power to the instrument Use of an adapter disconnects the utility ground creating a severe shock hazard Always connect the system power cord directly to an appropriate receptacle with a functional ground Power Supply Use only the power supply shipped with the instrument and operate it within the range of line voltages listed on it 1 Locate the ...

Page 38: ...14 Chapter 2 Installation Power cord left and power supply right are supplied Synergy LX User Manual 1501000N Revision A BioTek Instruments Inc ...

Page 39: ... is something you can fix do so now and run another system test If the problem is something you cannot fix or if the test continues to fail contact Technical Support Optional Press the lighted blue button on the front of the reader to store the microplate carrier in its chamber 7 Gen5 control only Start Gen5 and Test Communication 1 Start Gen5 2 If prompted to add a reader click Yes Otherwise sele...

Page 40: ...lification Your Synergy LX was fully tested at BioTek prior to shipment and should operate properly following the successful completion of the installation and setup procedures described in this chapter If you suspect that problems occurred during shipment if you received the reader back from BioTek following service or repair or if regulatory requirements dictate that Operational Performance Qual...

Page 41: ...tion instructions l Ensure the microplate carrier is empty Spilled fluids can contaminate the optics and damage the instrument l Install the shipping hardware see next section l The instrument s packaging design is subject to change If the instructions in this document do not apply to the packaging materials you are using contact Technical Support for guidance l Be sure to use packaging materials ...

Page 42: ...benchtop so you can easily access both the front and back panels 2 Remove the light blocking plug from the reader s back panel 3 Open the reader s front access door and o Insert the light blocking plug into its storage location o Using the supplied 3 32 hex wrench remove the shipping screw from its storage location Set it aside for a moment 4 If the microplate carrier is inside the reader press th...

Page 43: ...le holding the front end 2 or 3 inches above the benchtop insert the shipping screw inside the hole on the reader s back panel Without pushing against the carrier twist the screw clockwise until it catches the hole in the carrier block 9 Use the supplied 3 32 hex wrench to firmly seat the shipping screw Test this by tipping up the back of the reader If the carrier does not move the reader is ready...

Page 44: ...r 2 Tape the reader s front access door shut then place the reader inside the original plastic bag and carefully lower the reader onto the four foam cubes in the shipping container 3 Place the foam top tray on top of the reader in the shipping container 4 Place the accessory tray in the box and then place the accessories into the tray as shown in the figure 5 Close the top of the box and secure it...

Page 45: ...Repackaging and Shipping Instructions 21 Synergy LX User Manual 1501000N Revision A BioTek Instruments Inc ...

Page 46: ...22 Chapter 2 Installation Synergy LX User Manual 1501000N Revision A BioTek Instruments Inc ...

Page 47: ...nergy LX It also contains recommendations for optimum performance External Components 24 Filter Cubes 26 Operate the Reader Using the Touchscreen 34 Run a Take3 Session Using the Touchscreen 45 Getting Started with Gen5 Software 48 Recommendations for Optimum Performance 50 ...

Page 48: ... models 2 microplate carrier access door 3 power on off 4 carrier in out status LED 5 front compartment access door shown open with touchscreen behind it 1 USB port for printer 2 power inlet 3 USB port for host computer 4 shipping screw 5 fan filter access Synergy LX User Manual 1501000N Revision A BioTek Instruments Inc ...

Page 49: ...luorescence capable models 2 microplate carrier access door 3 power on off 4 carrier in out status LED 5 front compartment access door 1 power inlet 2 USB port for host computer 3 shipping screw 4 fan filter access Synergy LX User Manual 1501000N Revision A BioTek Instruments Inc ...

Page 50: ...ter cube installed to perform a fluorescence or luminescence read Configure a Filter Cube For models with fluorescence luminescence capability only Filter Cube F models The reader s internal filter cube table must exactly match the contents of the installed filter cube Gen5 users The Gen5 software filter cube table must exactly match the contents of the filter cube If you exchange the filter cube ...

Page 51: ...mpty 1505000 N A N A N A A filter cube must be configured in Gen5 or through the touchscreen as applicable prior to use Procedure If using Gen5 select System Optics Library Filter Cubes Click Help for guidance If using the touchscreen 1 From the Main Menu tap Instrument and tap the Filter Cubes tab 2 Enter a cube name It is recommended that you use the name on the filter cube s label 3 Toggle thro...

Page 52: ...en5 Optics Library or for models with a touchscreen the Filter Cubes tab on the Instrument Menu to identify and manage the contents of multiple filter cubes 1 Open the access door on the reader s front panel 2 Slide the filter cube into the filter cube port making sure the label on the filter cube handle is right side up See the photo below Synergy LX User Manual 1501000N Revision A BioTek Instrum...

Page 53: ...all See Configure a Filter Cube on page 26 Install Filters in an Empty Filter Cube Do not use a sharp instrument when removing or replacing a filter or C clip filter retainer Do not touch the filters or mirror with bare fingers Handle the mirror by its edges Synergy LX User Manual 1501000N Revision A BioTek Instruments Inc ...

Page 54: ...per l Linen or cloth gloves for handling the mirror Procedure 1 Remove the screw and EM filter holder 2 Using the following diagram install the apertures filters C clips and mirror Replace the EM filter holder when finished Synergy LX User Manual 1501000N Revision A BioTek Instruments Inc ...

Page 55: ...Filter Cubes 31 Synergy LX User Manual 1501000N Revision A BioTek Instruments Inc ...

Page 56: ...manual that ships in the printer s box for installation and setup instructions Load the Printer Paper 1 Ensure the printer is turned on 2 Press the gray tab on the right side of the printer to open the door 3 Place a roll of paper into the indented space on the back of the door with the leading edge of the paper coming from the top of the roll Ensure that the paper roll is installed so that the sh...

Page 57: ... not print check that the printer s power cable and USB cable are securely connected and run the test again l To print the results of a read tap Output in the top right corner of the results screen displayed when the read finishes l To print System Test results tap Print in the screen that appears when the system test finishes l To cleanly tear the paper from the printer pull upward from left to r...

Page 58: ...r check box or to activate a tab tap the item once l To return to the Main Menu from any other screen tap Home in the top left corner l For instructions on cleaning the touchscreen see page 55 l To preserve the life of the halogen lamp fluorescence turn it off when not needed by tapping in the Instrument Menu Main Menu The left side of the Main Menu lists the five protocols most recently created a...

Page 59: ...n Menu provides access to the following features l Quick Define and run an Absorbance Fluorescence or Luminescence read or perform a standalone shake Quick reads do not support shake or delay actions and they read the full plate See page 37 l Protocol Edit create and save delete and copy protocols Through this menu option protocols support shake and delay actions and at run time allow you to selec...

Page 60: ...any changes skip to the next topic Define Output Formats for Results Data on page 37 2 Tap in the Time field Tap the hour value and use the keypad to enter the correct time for both the hour and minutes then click OK 3 Tap the Date button 4 Tap month day or year and use the keypad to set the current date Tap Save when done 5 To set the Time Format Date Format Decimal Symbol and List Separator for ...

Page 61: ...value for each well after subtracting the average of the blank well s If you want to send results to a USB flash drive inserted in the reader on the Output tab toggle the USB Flash Drive button among the options l Report Generates a csv file containing the measurement values with Raw data Dual OD Blanked data as applicable This file can be opened in Excel or other spreadsheet software l Gen5 Input...

Page 62: ...e a Protocol The maximum number of uniquely named protocols that can be saved on the touchscreen at the same time is 60 excluding the predefined Take3 protocols 1 From the Main Menu tap Protocol select the desired detection method tab then tap Create 2 Use the onscreen keyboard to enter a name for the protocol then tap Save Note that the protocol name is limited to 18 characters 3 Define the proto...

Page 63: ... to change the view Primary and optional Secondary Tap the field to define the primary or secondary wavelength Enter a number within the valid range and tap Save Read Speed This field is set to Normal by default The other option is Sweep Refer to Appendix A to compare performance and timing specifications between the two options Activity Before Read Use the options here to include a delay up to 15...

Page 64: ... is initiated the reader performs an AutoScale routine see below to determine the optimum PMT gain setting for the wells to be measured Alternatively tap to change AutoScale to Use Value and then enter a specific gain number in the range of 25 255 If AutoScale is selected the reader measures the full plate or the columns selected at read time as fast as possible using the defined protocol paramete...

Page 65: ...e specific requirements The maximum allowed plate height for Synergy LX is 0 81 20 57 mm Dynamic Range The selection here 0 to 99 999 standard or 0 to 5 800 000 extended indicates the range over which the measured relative fluorescence units RFUs will be reported The greater the range the greater the separation between samples or standards Activity Before Read Use the options here to include a del...

Page 66: ...plate It is also stored with the results and will be included in the output to printer and or USB flash drive Integration Time By default the reader measures each well for 1 second collecting 50 samples which are then averaged The valid range is 0 02 1 sample to 99 90 seconds per well Increase the integration time for better precision decrease it to improve read speed Read Height This value repres...

Page 67: ...Note that only plate read results are saved System Test results are not saved they can only be exported to USB flash drive or printed The Synergy LX ships with several preprogrammed protocols Depending on the reader s model these protocols include Absorbance BCA and Bradford Fluorescence Red Blue and Green Luminescence Lum unfiltered and seven absorbance Take3 protocols 1 From the Main Menu tap a ...

Page 68: ...dit a Protocol 1 From the Main Menu tap Protocol 2 Tap the protocol that you want to modify then tap Edit 3 Make any desired changes then tap Save Delete a Protocol 1 From the Main Menu tap Protocol 2 Tap the protocol you want to delete then tap Delete Copy a Protocol Copying an existing protocol and then editing it is one way to create a new protocol 1 In the Main Menu tap the protocol you want t...

Page 69: ...t button to toggle through the results for each quadrant of the plate In the example below results for the lower right quadrant of the plate are displayed Run a Take3 Session Using the Touchscreen For models with absorbance capability only If you will set up and measure the Take3 plate using Gen5 refer to the user guide that shipped with the plate The Synergy LX ships with seven default Take3 prot...

Page 70: ...ely clean before running the alignment process Refer to the Take3 Take3 Trio User Guide for cleaning instructions If you remove or replace the glass slide perform the Pathlength Calibration procedure described in the Take3 Take3 Trio User Guide and then run the alignment procedure described below Update the Pathlength Values on the touchscreen with the new values generated through the Pathlength C...

Page 71: ...e the Take3 plate in the microplate carrier and tap Start l If the Take3 Blank Masking screen appears and reports that the CV Status is DIVERGING the CV of the blank wells is higher than the Blank threshold CV defined for the plate Invalid blank wells are highlighted in red Tap a well to mask exclude it from blank average and CV calculations When the remaining unmasked blank wells are valid the Ap...

Page 72: ...rotocol contains instructions for controlling the reader and optionally instructions for analyzing 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 Run the experiment to read plates and analyze the data To create a protocol 1 Optional but recommended Create...

Page 73: ...arameters l If the reader is connected and turned on Gen5 will communicate with the reader during this step to ensure that the protocol is valid l If the reader is not connected Gen5 will validate the syntax of the protocol for example the Read step is set between the Start and End Kinetic steps Later when you run the experiment Gen5 conducts another check and will present a message if any Gen5 pr...

Page 74: ...Automatic export to Microsoft Excel l Extensive data reduction options including cutoff and transformations l Ability to define standard wells and generates curves Recommendations for Optimum Performance General l Microplates should be clean and free from dust or bottom scratches Use new microplates from sealed packages Do not allow dust to settle on the surface of the solution use microplate cove...

Page 75: ...to help bring it within acceptable limits Use Tween 20 if possible or some other wetting agent to normalize the meniscus for absorbance measurements Some solutions develop menisci over a period of several minutes This effect varies with the brand of microplate and the solution composition As the center of the meniscus drops and shortens the light path the density readings change The meniscus shape...

Page 76: ...ons for cleaning and decontaminating the Synergy LX Maintenance Overview 53 Warnings and Precautions 54 Clean Exposed Surfaces 55 Clean the Touchscreen 55 Inspect Clean Filter Cubes 56 Clean the Air Filter 57 Replace the Halogen Bulb Assembly 59 Decontamination 60 ...

Page 77: ...ented in this schedule Task Page Frequency Clean External Surfaces page 55 As needed Inspect Clean Touchscreen page 55 As needed Inspect Clean the Filter Cubes page 56 As needed Clean the Air Filter page 57 As needed Replace the Halogen Bulb Assembly page 59 As needed Decontamination page 60 Before shipment or storage Required Materials l Mild detergent l Deionized or distilled water l Clean lint ...

Page 78: ...low water or other cleaning solution to run into the interior of the instrument If this happens contact Technical Support Do not soak the touchscreen Lubricants Do not apply lubricants to moving parts Lubricant on components in the carrier compartment will attract dust and other particles which may cause the instrument to produce an error Potential Biohazards Wear protective gloves when handling c...

Page 79: ...ater and mild detergent then thoroughly wring out the cloth so that liquid does not drip from it 3 Wipe the plate carrier the inside of the plate carrier door and front access door and all exposed surfaces of the instrument 4 If detergent was used wipe all surfaces with a cloth moistened with water 5 Use a clean dry lint free cloth to dry all wet surfaces If liquid is spilled inside the reader cal...

Page 80: ...t drip from it Do not soak the cloth 3 Wipe the touchscreen gently with the moist cloth 4 If detergent was used wipe the touchscreen with a cloth moistened with water 5 Dry the screen gently using another cloth Inspect Clean Filter Cubes For models with fluorescence luminescence capability only Filters should be inspected and cleaned at least every three months You ll need l Isopropyl ethyl or met...

Page 81: ...htly stroking its surface in one direction 4 Use a magnifying glass to inspect the surface for debris 5 When the filters are clean replace the filter cube and close the access door Clean the Air Filter 1 Using either a flat blade or 2 Phillips screwdriver remove the screw that secures the air filter cover on the rear of the reader Remove the cover and filter Synergy LX User Manual 1501000N Revisio...

Page 82: ...Rinse the air filter in water then allow it to dry completely 3 Replace the air filter and air filter cover on the reader and secure them in place with the screw Synergy LX User Manual 1501000N Revision A BioTek Instruments Inc ...

Page 83: ...ed 7 64 hex key unscrew the two screws that hold on the side door then remove the door and set it aside 3 Unplug the lamp s power plug and unscrew the thumbscrew that holds the lamp assembly in place Remove the lamp from the instrument 4 Slide the replacement lamp assembly into place using the lamp assembly s tabs as guides 5 Screw in the thumbscrew then finger tighten to secure the lamp Plug in t...

Page 84: ...strument Internal Voltage Always turn off the power switch and unplug the power supply before cleaning the outer surface of the instrument Liquids Do not immerse the instrument spray it with liquid or use a dripping wet cloth on it Do not allow water or other cleaning solution to run into the interior of the instrument If this happens contact Technical Support Touchscreen Avoid strong solvents suc...

Page 85: ... thoroughly wring it out so that liquid does not drip from it Do not soak the cloth 4 If the instrument is equipped with a touchscreen place the moistened cloth on the touchscreen and it let rest there for 15 minutes then remove it 5 Wipe the plate carrier and all exposed surfaces of the instrument including in the inside of the microplate carrier access door 6 Allow the instrument to dry for 20 m...

Page 86: ...the various test methods describes the materials and protocol parameters used to execute the tests explains how to analyze test results and provides troubleshooting tips in the event of a failure Instrument Qualification Procedures starting on page 85 contains the actual step by step test procedures System Test 63 Plate Shaker Test 63 Absorbance Testing 64 Fluorescence Testing 73 Luminescence Test...

Page 87: ...p the beeping If necessary initiate another system test to try to retrieve more information from the reader Refer to Error Conditions starting on page 119 for more information Refer to Sample Reports starting on page 113 to see a sample System Test Report for Synergy LX Plate Shaker Test This test verifies that the multi speed plate shaker is operating properly The test involves creating and runni...

Page 88: ... verify mechanical alignment Absorbance Test Plate PN 7260551 uses NIST traceable neutral density filters to confirm absorbance specifications in the UV range 340 nm Every test plate comes with a Test Plate Calibration Certificate containing a table with Absorbance OD Standards for each filter at each wavelength supported by the plate Before the Absorbance Plate Test can be performed the OD Standa...

Page 89: ...ished Once it is proven that the reader is accurate at these OD values the reader is also considered to be linear To further verify this you can perform a linear regression analysis on the test plate OD values in a program such as Microsoft Excel an R2 value of at least 0 9900 is expected l Repeatability This test ensures the instrument meets its repeatability specification by conducting repeated ...

Page 90: ...perly seated in the microplate carrier l Check the four alignment holes A1 A12 H1 H12 to ensure they are clear of debris l Check the microplate carrier to ensure it is clear of debris Accuracy Test l Check the neutral density filters to ensure they are clean positions C1 D4 E2 F5 G3 H6 If needed clean the filters with lens paper Do not remove any filters and do not use alcohol or other cleaning ag...

Page 91: ... test the instrument s alignment repeatability and accuracy by preparing a series of solutions of varying OD values as described on page 94 Absorbance Liquid Test 3 is an optional test offered for those sites that must have proof of linearity at 340 nm Alternatively the BioTek 340 nm Absorbance Test Plate may be used see page 64 This test is optional since the Synergy LX has good front end lineari...

Page 92: ...late Out In step between loops Text Please rotate the plate 180 degrees Synergy LX Abs Test 2 prt Parameter Setting Plate Type 96 WELL PLATE Two Read Steps Detection Method Absorbance Read Type Kinetic Kinetic loop one per Read step Set a Run Time Interval combination to read the plate five times with minimal delay Step labels First Read step Normal Second Read step Turnaround Read wells Full plat...

Page 93: ...n on page 92 All tests are conducted using the Normal read speed Absorbance Liquid Test 1 Accuracy Specification 1 0 0 010 OD from 0 000 to 2 000 OD 3 0 0 010 OD from 2 000 to 2 500 OD Repeatability Specification 1 0 0 005 OD from 0 000 to 2 000 OD 3 0 0 005 OD from 2 000 to 2 500 OD 1 The plate is read five times in the Normal position at 405 nm Calculate the Mean OD and Standard Deviation of tho...

Page 94: ...racy of 1 0 0 010 OD then the expected range for the Mean of the well in its Turnaround H12 position is 1 873 to 1 931 OD 1 902 x 0 010 0 010 0 029 1 902 0 029 1 873 1 902 0 029 1 931 Absorbance Liquid Test 2 Accuracy Specification 1 0 0 010 OD from 0 000 to 2 000 OD 3 0 0 010 OD from 2 000 to 2 500 OD Repeatability Specification 1 0 0 005 OD from 0 000 to 2 000 OD 3 0 0 005 OD from 2 000 to 2 500...

Page 95: ... l Compare the mean reading for well A1 to its mean reading when in the H12 position Next compare the mean values for the H1 well to the same well in the A12 position The difference in the values for any two corresponding wells should be within the Accuracy specification for 96 well plates If the four corner wells are within the accuracy range the reader is in alignment Example If the mean of well...

Page 96: ...ean OD values are the Y values The solution concentrations are the X values 1 00 0 75 0 50 Since it is somewhat difficult to achieve high pipetting accuracy when conducting linear dilutions an R2 value of at least 0 9900 is considered adequate Troubleshooting If an absorbance liquid test fails try the following If a test continues to fail contact Technical Support l Check the microwells and plate ...

Page 97: ...e with the test plate The protocols include embedded Microsoft Excel spreadsheets to automatically calculate results and determine pass fail The protocols and their spreadsheets were fully validated in accordance with BioTek Instruments Product Validation policies and procedures The package also contains a user guide that describes the test methods helps you get started with using the plate and pr...

Page 98: ...f using serial dilutions The tests presented in this section require specific microplates solutions wavelengths mirrors and filters Your laboratory may require a deviation from some of these tests For example you may wish to use a different fluorescing solution or microplate If deviation from the tests as presented in this section is required the following steps should be taken the first time each...

Page 99: ... Filter Sets Filter set Green Gain Auto Scale to High Wells D7 50000 Read Speed Normal Delay after plate movement 350 msec Measurements per data point 10 Dynamic Range Standard Read Height 8 75 mm Read Step 2 Kinetic Kinetic loop Run Time 0 01 35 Interval 0 00 06 Reads 16 Step Label Sensitivity Read Buffer Read Wells C9 E9 Filter Sets Filter set Green Gain Auto Use first filter set gain from FIRST...

Page 100: ...Label Linearity Read Read Wells C1 F5 Filter Sets Filter set Green Gain Auto Scale to High Wells C1 50000 Read Speed Normal Delay after plate movement 350 msec Measurements per data point 10 Dynamic Range Standard Read Height 7 75 mm Synergy LX_FI_T_MUB prt Parameter Default Setting Plate Type Costar 96 well black opaque 3915 Detection Method Fluorescence Three Read Steps Read Step 1 Kinetic Kinet...

Page 101: ...Read Buffer Read Well C9 D9 E9 Filter Sets Filter Set Blue Gain Auto Use first filter set gain from FIRST Read Step Read Speed Normal Delay after plate movement 350 msec Measurements per data point 10 Dynamic Range Standard Read Height 8 25 mm Read Step 3 Endpoint Step Label Linearity Read Read Well C1 F5 Filter Sets Filter Set Blue Gain Auto Scale to High Wells C1 80000 Read Speed Normal Delay af...

Page 102: ...ate the Mean for the 16 reads of the SF or MUB Concentration well D7 4 Calculate the Signal to Noise Ratio SNR using the Mean SF or MUB Concentration Buffer Media STD with its corresponding Buffer Mean SF or MUB Mean Buffer Mean 3 Buffer STD 5 Calculate the Detection Limit Sodium Fluorescein Using the known concentration value of SF and the calculated SNR 1000 SNR Filter Cube To pass the Detection...

Page 103: ...100 Mean of the 100 nM wells 50 Mean of the 50 nM wells 25 Mean of the 25 nM wells 12 5 Mean of the 12 5 nM wells 6 25 Mean of the 6 25 nM wells 3 Calculate the R2 value it must be 0 9500 to pass Troubleshooting If any tests fail please try the following suggestions If the test s continue to fail print the results and contact Technical Support l Are the solutions fresh Discard the plate and any op...

Page 104: ...he value is less than 30 o The stock solution dilution concentrations may be too high Try creating fresh solutions dilutions and rerun the test using a new clean plate o If all of the tests are passing but the Gain value is low a PMT in your reader may just be very sensitive Contact Technical Support to confirm that this may be the case If the value is greater than 200 o The stock solution dilutio...

Page 105: ...a known correlation of 35 photons per attomole of ATP By using the NIST data provided with the Harta plate in photons s a conversion factor of 0 02884 attomole photon is applied to determine an ATP concentration and subsequent limit of detection for the instrument under test Plate Layout Results Analysis The Luminescence Test procedure is described on page 106 1 Determine if the plate s battery is...

Page 106: ...is equipped with the red shifted PMT the detection limit must be 500 amol to pass If you do not know which PMT is installed 49984 low noise PMT 49721 red shifted PMT please contact Technical Support Gen5 Protocol Reading Parameters The information in this section represents the recommended reading parameters for the referenced Gen5 protocol s Synergy LX_LumTest_Harta prt Parameter Default Setting ...

Page 107: ...D1 G4 Filter Set LUM Gain 150 Integration Time 0 10 00 MM SS ss Delay After Plate Movement 350 msec Dynamic Range Standard Read Height 8 5 mm Read Step 3 Detection Method Luminescence Read Type Endpoint Read Wells A7 A8 Step Label Battery Check Filter Set LUM Gain 50 Integration Time 0 01 00 MM SS ss Delay After Plate Movement 350 msec Dynamic Range Extended Read Height 8 5 mm Synergy LX User Manu...

Page 108: ... Ensure that the reading is performed through a hole in the filter cube not through a glass filter l Verify that the filter cube settings in Gen5 match the filter cube installed in the reader l If the test continues to fail the optics block may need to be cleaned Contact Technical Support for instructions Synergy LX User Manual 1501000N Revision A BioTek Instruments Inc ...

Page 109: ... page 62 introduces the various test methods describes the materials and relevant protocol parameters used to execute the tests explains how to analyze test results and provides troubleshooting tips in the event of a failure Overview 86 IQ OQ PQ Description 86 Recommended Qualification Schedule 88 System Test 89 Absorbance Plate Test 90 Absorbance Liquid Tests 92 Fluorescence Plate Tests 98 Fluore...

Page 110: ...et your site s regulatory requirements A Product Qualification Package PN 1500534N for the Synergy LX is available for purchase The package contains complete procedures Gen5 protocols checklists and logbooks for performing Installation Qualification Operational Qualification Performance Qualification and Maintenance Contact your local BioTek dealer for more information IQ OQ PQ Description Install...

Page 111: ...arable 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 requirements of the tests performed at your laboratory l The recommended PQ procedure consists of performing the system test and a series of tests for the absorbance fluorescence and luminescence sys...

Page 112: ...ly Annually Monthly Quarterly All models Installation setup and configuration of the reader and if used host computer and Gen5 software ü System Test ü ü ü Models with absorbance capability Absorbance Plate Test ü ü Absorbance Liquid Test 1 or Liquid Test 2 ü ü Optional Absorbance Liquid Test 3 or 340 nm Absorbance Plate Test using PN 7260551 ü ü Models with fluorescence capability Corners Sensiti...

Page 113: ... the Gen5 Close the box the test report contains the error code that was generated by the failure 2 When the test is complete a dialog appears requesting additional information Enter any required information and then click OK 3 The results report appears it shows either SYSTEM TEST PASS or SYSTEM TEST FAIL ERROR error code DETECTED If the test failed look up the error code in Appendix B Error Cond...

Page 114: ...e current Test Plate Calibration Certificate 2 Start Gen5 and select System Diagnostics Test Plates Add Modify Plates 3 Click Add The Absorbance Test Plate dialog appears 4 Select the appropriate Plate Type and then enter the plate s serial number 5 Enter the Last Certification and Next Certification dates from the calibration label on the Test Plate 6 If the wavelength values in the top row of th...

Page 115: ...ed to select only those wavelengths most appropriate for your use of the reader 4 Optional Enter a comment 5 Click Start Test 6 Place the Absorbance Test Plate on the microplate carrier with well A1 in the proper location 7 Click OK to run the test 8 When the test is complete the results report appears Scroll through the report every result should show PASS l Troubleshooting tips are provided on p...

Page 116: ... comparable then the results from these tests will be your baseline for future tests Document your new test procedure and save all test results Absorbance Liquid Test 1 Materials Manufacturer part numbers are subject to change l New 96 well clear flat bottom microplate Corning Costar 3590 recommended l Stock Solution A or B which may be formulated by diluting a dye solution available from BioTek A...

Page 117: ...ed stock solution Solution A or B prepare a 1 2 dilution using deionized water one part stock one part deionized water the resulting solution is a 1 2 dilution The concentrated stock solution should have an optical density of approximately 2 000 OD or lower 2 Pipette 200 µL of the stock solution into column 1 3 Pipette 200 µL of the diluted solution into column 2 After pipetting the diluted test s...

Page 118: ...l clear flat bottom microplate Corning Costar 3590 is recommended l Ten test tubes numbered consecutively set up in a rack l Calibrated hand pipette Class A volumetric pipette recommended l Solution A or B see the instructions for Liquid Test 1 l A 0 05 solution of deionized water and Tween 20 l The Gen5 protocol Synergy LX Abs Test 2 prt described on page 68 Test Procedure 1 Create a percentage d...

Page 119: ...he remaining tubes into the wells of the corresponding column of the microplate Tube 2 into wells A2 to H2 Tube 3 into wells A3 to H3 and so on After pipetting the diluted test solution into the microplate and before reading the plate we strongly recommend shaking the plate for four minutes This will allow any air bubbles in the solution to settle and the meniscus to stabilize Alternatively wait 2...

Page 120: ...duated cylinder l Precision balance with readability to 0 01 g l Buffer solution described below l The Gen5 protocol Synergy LX Abs Test 3 prt described on page 69 Buffer Solution l Deionized water l Phosphate Buffered Saline PBS pH 7 2 7 6 Sigma tablets P4417 or equivalent l β NADH Powder β Nicotinamide Adenine Dinucleotide Reduced Form Sigma bulk catalog number N 8129 or preweighed 10 mg vials S...

Page 121: ...ls of column 5 and 6 After pipetting the diluted test solution into the microplate and before reading the plate we strongly recommend shaking the plate for four minutes This will allow any air bubbles in the solution to settle and the meniscus to stabilize Alternatively wait 20 minutes after pipetting the test solution before reading the plate 4 Create a Gen5 experiment based on the Synergy LX Abs...

Page 122: ...protocols on the USB flash drive that came with the test plate Use only those protocols that apply to your reader model and your organization s qualification requirements Fluorescence Liquid Tests Fluorescence Liquid Tests starting on page 74 describes the test methods lists the Gen5 protocol parameters explains how to analyze the test results and provides troubleshooting tips in the event of test...

Page 123: ...ergy LX FI_T_SF prt o Synergy LX FI_T_MUB prt Corners Sensitivity Linearity FI Tests Manufacturer part numbers are subject to change The materials listed here are for use with Sodium Fluorescein Methylumbelliferone can be used as an alternate or supplemental method for performing these tests See page 102 If using test kit PN 7160013 see page 5 the buffer and SF are pre diluted l Buffer o NIST trac...

Page 124: ... solution in foil to prevent exposure to light Discard any open unused solution after seven days 1 The Sodium Borate solution does not require further preparation proceed to step 2 If you are using PBS prepare the solution l Optional but recommended Using a 0 45 micron filter filter 200 mL of deionized or distilled water l Follow the manufacturer s instructions on the PBS packaging to create 200 m...

Page 125: ...ge 74 4 Read the plate and then save the experiment Pipette Map Seal the plates with foil or store them in black polyethylene bags until use When using a clear bottom plate if the base of the plate is touched clean the entire base with alcohol 95 ethanol and then wipe with a lint free cloth Before placing the plate in the instrument blow the bottom of the plate with an aerosol duster Corners Sensi...

Page 126: ...s l Aspirate 150 μL from column 3 and dispense into column 4 Mix the wells using the pipette Do not discard the tips l Aspirate 150 μL from column 4 and dispense into column 5 Mix the wells using the pipette Do not discard the tips l Aspirate 150 μL from column 5 and discard the tips Alternate Supplemental Tests Using Methylumbelliferone MUB As an alternative to using Sodium Fluorescein Methylumbe...

Page 127: ...nded 0 45 micron filter l Optional Black polyethylene bag s to temporarily store plate s l Gen5 protocol described on page 74 o Synergy LX FI_T_MUB prt Test Solutions Filter solutions to remove particulates that could cause erroneous readings Do not allow dust to settle on the surface of the solution use microplate covers or seals when not reading the plate Wrap the vial containing the MUB stock s...

Page 128: ... mL of 1 mg mL solution 4 12 mL of CBB 176 μg mL 0 1 mL of 176 μg mL solution 9 9 mL of CBB 1 76 μg mL 0 5 mL of 1 76 μg mL solution 4 5 mL of CBB 176 ng mL 1 mL of 176 ng mL solution 9 mL of CBB 17 6 ng mL 100 nM Test Procedure 1 If you have not already done so prepare the test solutions See page 103 2 Perform the Sensitivity and Linearity tests l Pipette the solutions into a clean 96 well plate ...

Page 129: ...h just four tips installed l Pipette 150 µL of buffer into columns 2 5 not column 1 Discard the tips l Pipette 150 µL of the 17 6 ng mL 100 nM solution into column 1 Discard the tips l Pipette 150 µL of the 17 6 ng mL 100 nM solution into column 2 Do not discard the tips l Aspirate 150 µL from column 2 and dispense it into column 3 Mix the wells using the pipette Do not discard the tips l Aspirate...

Page 130: ... Microplate PN 8030015 which includes microplate carrier adapter PN 8032028 l Gen5 protocol Synergy LX_LumTest_Harta prt described on page 82 l LUM filter cube EX Plug EM Hole Mirror Empty Test Procedure 1 Turn on the Harta reference plate using the I O switch on the back of the plate 2 Check the plate s battery by pressing the test button on the back of the plate and ensuring that the test light ...

Page 131: ...hen place the test plate on top of the adapter 4 Create a Gen5 experiment based on the Synergy LX_ LumTest_Harta protocol described on page 82 and read the plate 5 When the experiment is complete calculate and evaluate results as described under Results Analysis on page 81 6 When finished turn off the Harta reference plate to preserve battery life Synergy LX User Manual 1501000N Revision A BioTek ...

Page 132: ... Specifications This appendix contains BioTek s published specifications for the Synergy LX General Specifications 109 Absorbance Specifications 110 Fluorescence Specifications 112 Luminescence Specifications 112 ...

Page 133: ...bulb Fluorescence reads halogen bulb 500 hour lifespan Dimensions Touchscreen models 15 H x 15 W x 15 D 38 1 x 38 1 x 38 1 cm Non touchscreen models 12 H x 15 W x 15 D 30 5 x 38 1 x 38 1 cm Weight 27 lbs 12 3 kg Environment Operational temperature range 18 C to 37 C 64 4 F to 98 6 F Storage temperature range 25 C to 50 C Humidity Operational 10 to 85 relative humidity non condensing Storage 10 to ...

Page 134: ...econds 96 well microplate Throughput time elapsed from plate moving in to plate moving out 96 Well 450 nm sweep read mode 35 seconds Accuracy Tested with certified neutral density glass 96 well plate normal read speed 0 000 to 2 000 OD 1 0 0 010 OD delay after plate movement 100 ms 2 000 to 2 500 OD 3 0 0 010 OD delay after plate movement 100 ms 384 well plate normal read speed 0 000 to 1 500 OD 2...

Page 135: ...ity Standard Deviation STD Tested with certified neutral density glass measured by one standard deviation 8 measurements data point 96 well plate normal read speed 0 000 to 2 000 OD 1 0 0 005 OD delay after plate movement 100 ms 2 000 to 2 500 OD 3 0 0 005 OD delay after plate movement 100 ms 384 well plate normal read speed 0 000 to 1 500 OD 1 0 0 005 OD delay after plate movement 100 ms 1 500 to...

Page 136: ...owing read settings Delay after plate movement 100 ms Measurements per data point 10 Wavelength range 320 700 nm low noise PMT 320 850 nm red shifted PMT Luminescence Specifications Production testing is performed using a Harta plate 96 well plate with standard low noise PMT Detection Limit for ATP 75 amol well Integration Time 10 seconds PMT Gain 150 Blank Wells 16 Wavelength Range 200 700 nm 96 ...

Page 137: ...Appendix B Sample Reports This appendix contains sample System Test and Absorbance Plate Test reports for the Synergy LX ...

Page 138: ...114 Appendix B Sample Reports Sample System Report Synergy LX User Manual 1501000N Revision A BioTek Instruments Inc ...

Page 139: ...Sample System Report 115 Synergy LX User Manual 1501000N Revision A BioTek Instruments Inc ...

Page 140: ...116 Appendix B Sample Reports Sample Absorbance Test Plate Report Synergy LX User Manual 1501000N Revision A BioTek Instruments Inc ...

Page 141: ...Sample Assay Results 117 Sample Assay Results Sample Report CSV File Synergy LX User Manual 1501000N Revision A BioTek Instruments Inc ...

Page 142: ...118 Appendix B Sample Reports Sample Gen5 input TXT File Synergy LX User Manual 1501000N Revision A BioTek Instruments Inc ...

Page 143: ...Appendix C Error Conditions This appendix describes error conditions and provides troubleshooting tips Overview 120 Error Codes 120 ...

Page 144: ...rror code appears you should run a system test for diagnostic purposes See System Test on page 89 Having the system test report before calling the Technical Support can speed the resolution of the error If an error message appears during a plate read it is the user s responsibility to determine the validity of any data that was received Use this appendix to diagnose problems and solve them if poss...

Page 145: ...dware module is not available A301 The y axis hardware module is not available A302 The z axis hardware module is not available A303 The order sorting filter wheel hardware module is not available A304 The monochromator hardware module is not available A500 EEProm memory read write error AB00 Multiprocessor error AC00 External RAM error ADXX Stack error AEXX Invalid motor ramp profile 7FXX Dual pr...

Page 146: ...alog digital data collection failure A D converter for measurement circuits has failed or is generating inconsistent or noisy data For Fluorescence measurements Absorbance measurements Voltage reference measurements 0401 Fluorescence measurement data converter never indicated ready Turning the instrument off and on may resolve the problem If the error reoccurs contact Technical Support 0411 Fluore...

Page 147: ...ng the system self test 0600 24 VDC is outside of limits Turning the instrument off and on may resolve the problem If the error reoccurs contact Technical Support 0610 24 VDC is noisy or inconsistent 0601 5 VDC is outside of limits 0611 5 VDC is noisy or inconsistent 0602 Lamp current test Lamp current is outside of limits 0612 Lamp current test Lamp current is noisy or inconsistent 0603 Lamp off ...

Page 148: ...Flash voltage is outside limits 0617 Mono flash absorbance high power test Flash voltage is noisy or inconsistent 0608 Mono flash absorbance maximum power test Flash voltage is outside limits 0618 Mono flash absorbance maximum power test Flash voltage is noisy or inconsistent 08XX errors Configuration checksum errors 0800 0801 0802 Processor EEPROM configuration memory checksum mismatch Basecode m...

Page 149: ...nes which may corrupt data being sent Perform a global reboot as described above If the error reoccurs contact Technical Support 21XX errors Invalid parameter error This error will occur when the instrument is sent protocol parameters that are incompatible with it Because Gen5 checks most of these parameters against the instrument model when the protocol is being defined these errors generally ser...

Page 150: ...r incorrectly formatted XML file 2104 Filter mirror block invalid excitation wavelength 2106 Filter mirror block invalid emission wavelength 2101 Probe Z not top fluorescence probe 2101 Set lamp state state out of range 2101 Invalid dip switch selection 2101 Flash and shake counter invalid counter 2101 Autosensitivity invalid event number or event is not a read 2102 Invalid dip switch state 2102 A...

Page 151: ... shake motor 2 must be x or y axis 22XX errors Hardware mismatch error Autocalibration is being requested for an instrument configuration that does not exist 2201 Carrier to fluorescence probe If error is repeatable contact Technical Support 2202 Carrier to absorbance probe 2203 Carrier sensor positions 2204 Probe Z 2205 Carrier Z tilt correction 23XX errors Item not found error This indicates tha...

Page 152: ...selected plate Specific error indicates which dimension is incorrect 2403 Invalid first row dimension Verify that dimensions for specific plate are correctly defined in Gen5 plate definition table 2404 Invalid last row dimension 2405 Invalid first column dimension 2406 Invalid last column dimension 2407 Invalid plate width 2408 Invalid plate length 2409 Invalid plate height 2500 error Lamp off err...

Page 153: ...herwise the process waits for the read setup time to arrive Verify computer set up Hibernate and or sleep mode cannot be enabled 2D32 Read event expected Repeat shake must be followed by read 2D33 Invalid shake time Invalid shake time selected 2D34 Invalid shake speed Invalid shake speed selected 2D42 Invalid delay time Invalid time selected for delay event 2D45 Spectral scan increment Invalid inc...

Page 154: ...te specification in user manual and move to area of lower humidity 3710 Absorbance measurement channel failed noise test 38XX errors Absorbance fixed offset test failed 3800 Absorbance reference channel failed offset test Too much light in read chamber Ensure that instrument enclosure is completely installed and secured Ensure the carrier door is closing fully Ensure the filter cube access door is...

Page 155: ...ntact Technical Support 3A1X Absorbance gain out of range where x optics test wavelength number 3EXX errors Absorbance read saturated Fail if measurement is 65535 Either absorbance reference or measurement channel has saturated 3E10 Absorbance measurement channel saturation during carrier auto calibration when on solid area of jig The carrier is moved off the center region and absorbance integrati...

Page 156: ...ion Check for spill in chamber Contamination on the lower or the upper optical assembly in the absorbance light path can cause this error A system test will typically show elevated gains in the 230 nm wavelength if the absorbance optics is contaminated Contact Technical Support Note This error may not occur during a system test but can occur when the first read is attempted 3F0Yw Yw wavelength rea...

Page 157: ...n chamber 3F11 Make sure any lids or seals used are not blocking the light path hole at left rear of carrier Ensure that the door is completely closed Ensure that instrument enclosure is completely installed and secured Flash Lamp missing flashes or too much flash to flash intensity variation If repeatable contact Technical Support 3F11 Possible light blockage on rear of carrier over measurement v...

Page 158: ...well plate Well location error code for 24 well plate Well location error code for 48 well plate Note Follow the same patterns for 96 well and 384 well plates Sensitivity too high Chemistry too concentrated hot Verify that the physical filter configuration matches Gen5 filter table 47XX errors PMT Fluorescence noise test error Synergy LX User Manual 1501000N Revision A BioTek Instruments Inc ...

Page 159: ...ured If repeatable contact Technical Support 4810 PMT measurement offset saturation test 4AXX errors Fluorescence gain error 4A00 PMT gain out of range Too much light in chamber Ensure that instrument case is completely installed and secured If repeatable contact Technical Support 4A1Y A flash fluorescence read error is flagged 4A1y y readset if the current PMT sensitivity does not match the sensi...

Page 160: ...ier enters chamber Sensitivity set to high try adjusting auto sensitivity setting Door not closing completely If repeatable contact Technical Support 4E1X PMT measurement channel saturation 4FXX errors Fluorescence signal out of range 4F00 Fluorescence reference signal out of range Sensitivity too high Ensure that door is fully closed Ensure instrument cover is installed and secured Open hole in e...

Page 161: ...stalled If the error occurs on a carrier in move the middle sensor is responding as if it is already closed a disconnected or defective opto sensor responds in this manner An object may be obstructing the path If repeatable contact Technical Support 5001 5201 Carrier Y failed to home Shipping bracket installed Y axis rails are dirty or rusty where the dirt in roller bearing is causing bearings to ...

Page 162: ...omator failed autocalibration 540X errors Verify movements 5400 Carrier X failed positional verify Verify there are no obstructions and that the carrier and the dispenser probe Z Axis are not hitting anything If repeatable contact Technical Support 5401 Carrier Y failed positional verify 5402 Probe Z Axis failed positional verify 5404 Monochromator failed positional verify If repeatable contact Te...

Page 163: ...definition dimensions are correct especially for custom defined plates Verify plate height matches selected plate type If repeatable or reoccurs contact Technical Support 5701 Carrier Y Axis move attempted outside physical limits of travel 5702 Vertical Z Axis move attempted outside physical limits of travel 5704 Monochromator not homed successfully 5800 Carrier X alternate sensor error Verify the...

Page 164: ...Z Axis probe told to move to position that would result in hitting plate This error can occur if the carrier is inside and the newly defined plate height is different from the most recently specified plate height To resolve this error eject the carrier prior to running the experiment Plate is inside chamber when it should be outside This may occur if read was aborted and home all axis not performe...

Page 165: ...Verify that EX filter configuration table is set correctly 2112 Invalid EX filter bandwidth EX filter bandwidth does not match available filters Verify that EX filter configuration table is set correctly 2113 Invalid EM filter wavelength EM filter wavelength does not match available filters Verify that EM filter configuration table is set correctly 2114 Invalid EM filter bandwidth EM filter bandwi...

Page 166: ...e empty Enter protocol name 4040 Blanks defined that are beyond the limits of the plate Ensure that the number of blanks does not exceed the total number of wells on the plate 4050 No compatible filters configured Filters in protocol are not configured in reader Verify that correct filters have been installed and update filter configuration table Note 50XX errors indicate problems with a device co...

Page 167: ...ectory 5016 USB port Open file error 5017 USB port Close file error 5018 USB port Error changing directory 5019 USB port Error writing to printer 501A USB port Error deleting file 501B USB port Error syncing 501C USB port No output destination selected 501D USB port No flash drive detected 7FDD Take3 plate no longer supported In order to create and save a Take3 protocol the S N must be entered and...

Page 168: ...eed replacement See users guide for instructions 7FE0 Error parsing assay response If repeatable or reoccurs contact Technical Support 7FF2 Error calibrating Take3 plate 7FF3 Error determining bulb status 7FF4 Wells over range while determining AutoScale Wells are out of range probably saturated The reader will lower gain values for a 2nd try but if wells are still out of range this error will res...

Page 169: ...cted ETX missing 7FF9 Checksum error parsing assay response 7FFA Error parsing reader response 7FFB Error parsing assay response 7FFC Error waiting to send command 7FFD Checksum error parsing command response 81XX Communication error A200 Basecode versions do not match AC00 External RAM error Synergy LX User Manual 1501000N Revision A BioTek Instruments Inc ...

Page 170: ...iligheidsmededelingen Avis de sécurité Sicherheitshinweise Avvisi di sicurezza Avisos de seguridad This appendix contains safety information for the Synergy LX translated into Dutch French German Italian and Spanish Safety Notices 147 ...

Page 171: ...productdocumentatie Portez une attention particulière aux avis de sécurité suivants dans l ensemble de la documentation du produit Achten Sie besonders auf die folgenden Sicherheitshinweise in allen Produktdokumentationen Prestare particolare attenzione agli avvisi di sicurezza presenti in tutta la documentazione del prodotto Preste especial atención a los siguientes avisos de seguridad en toda la...

Page 172: ...onditions indiquées sont entièrement comprises et remplies Ein WARNHINWEIS weist auf eine Gefahr hin Er weist auf ein Betriebsverfahren eine Vorgehensweise oder ähnliches hin deren falsche Ausführung oder Nichtbeachtung zu Verletzungen oder zum Tod führen können Fahren Sie bei einem WARNHINWEIS erst dann mit Ihrer Arbeit fort wenn die angegebenen Bedingungen vollständig verstanden und erfüllt sind...

Page 173: ...ns indiquées sont entièrement comprises et remplies Ein VORSICHTSHINWEIS weist auf eine Gefahr hin Er weist auf ein Betriebsverfahren eine Vorgehensweise oder ähnliches hin deren falsche Ausführung oder Nichtbeachtung zu einer Beschädigung des Produkts oder zum Verlust wichtiger Daten führen kann Fahren Sie bei einem VORSICHTSHINWEIS erst dann mit Ihrer Arbeit fort wenn die angegebenen Bedingungen...

Page 174: ...ésactivez toujours l interrupteur d alimentation électrique et débranchez l alimentation avant de nettoyer la surface extérieure de l instrument Spannung im Geräteinneren Vor dem Reinigen der Außenfläche des Geräts grundsätzlich den Stromschalter ausschalten und das Stromkabel aus der Steckdose ziehen Tensione interna Spegnere sempre l interruttore dell alimentazione e scollegare l alimentazione p...

Page 175: ...entraîner un choc électrique et un risque d incendie Leistungsbemessung Die Stromversorgung des Geräts bzw das Anschlusskabel muss mit einer Steckdose verbunden werden deren Spannungs und Stromwerte innerhalb der für das System vorgeschriebenen Nennwerte liegen Die Verwendung einer nicht kompatiblen Steckdose kann zu einem elektrischen Schlag und Brandgefahr führen Potenza nominale L alimentazione...

Page 176: ...riée dotée d une mise à la terre fonctionnelle Elektrische Erdung Verwenden Sie niemals einen Steckeradapter zum Anschließen der Primärstromversorgung an die externe Stromversorgung Bei Verwendung eines Adapters wird die Verbindung zur Gebäudeerde unterbrochen sodass ein erhebliches Stromschlagrisiko besteht Das Stromkabel ist immer direkt an eine geeignete Steckdose mit Funktionserdung anzuschlie...

Page 177: ... the power supply shipped with the instrument and operate it within the range of line voltages listed on it Voeding Gebruik alleen de voeding die bij het instrument is geleverd en gebruik deze binnen het bereik van de netspanningen die op de voeding staan vermeld Alimentation électrique Utilisez exclusivement l alimentation électrique fournie avec l instrument dans la plage de tension de ligne ind...

Page 178: ...iligheidsbril en geschikte beschermingsmiddelen zoals chemicaliënbestendige rubberen handschoenen en een schort Risques biologiques potentiels Certains tests ou échantillons peuvent présenter un risque biologique Des précautions de sécurité adéquates doivent être prises comme indiqué dans la notice de l emballage du test Portez toujours des lunettes de sécurité et un équipement de protection appro...

Page 179: ...zione appropriati ad esempio guanti e grembiule in gomma resistenti alle sostanze chimiche Riesgos biológicos potenciales Algunos ensayos y especímenes pueden constituir un riesgo biológico Se han de tomar precauciones de seguridad suficientes tal como se indica en el folleto del paquete del ensayo Use siempre gafas de seguridad y equipos protectores adecuados como guantes de caucho resistentes a ...

Page 180: ...s composants internes ont été exposés à du liquide Flüssigkeiten Keine Flüssigkeiten auf dem Gerät verschütten In die Bauteile im Geräteinneren bilden einsickernde Flüssigkeiten ein Potenzial für die Gefahr von Stromschlägen oder Schäden am Gerät Bei Verschütten von Flüssigkeiten während ein Programm läuft ist dieses zu stoppen und das Gerät auszuschalten Verschüttete Flüssigkeiten sind unverzügli...

Page 181: ...héant contactez l assistance technique N immergez pas l écran tactile Flüssigkeiten Das Gerät nicht in Flüssigkeit eintauchen oder damit einsprühen und keine tropfnassen Tücher verwenden Kein Wasser oder andere Reinigungslösung in das Geräteinnere eindringen lassen Sollte dies vorkommen setzen Sie sich mit dem technischen Kundendienst in Verbindung Den Touchscreen nicht einweichen Liquidi Non imme...

Page 182: ...les températures fluctuent au dessus ou au dessous de cette plage Umgebungsbedingungen Das Gerät darf keinen Extremtemperaturen ausgesetzt werden Für den ordnungsgemäßen Betrieb müssen die Temperaturen in Gerätenähe in den im Abschnitt Spezifikationen dieses Dokuments angegebenen Grenzen bleiben Temperaturschwankungen über diese Grenzwerte hinaus können die Geräteleistung beeinträchtigen Condizion...

Page 183: ...trument Veillez à rincer et essuyer soigneusement toutes les surfaces Natriumhypochlorit Kein Teil des Geräts darf der empfohlenen verdünnten Natriumhypochloritlösung länger als 20 Minuten lang ausgesetzt werden Bei längerem Kontakt drohen Beschädigungen an den Geräteoberflächen Alle Oberflächen unbedingt abspülen und gründlich abwischen Ipoclorito di sodio Non esporre nessun componente dello stru...

Page 184: ...ttire la poussière et autres particules ce qui peut provoquer une erreur de l instrument Schmierstoffe Keine Schmierstoffe auf bewegliche Teile auftragen Schmierstoffe auf Komponenten im Trägerfach ziehen Staub und andere Teilchen an die zu einem Gerätefehler führen können Lubrificanti Non applicare lubrificanti alle parti in movimento La presenza di lubrificante sui componenti del vano portapiast...

Page 185: ...laissez l ampoule refroidir pendant 15 minutes au moins avant de la remplacer Heiße Oberflächen Bei eingeschaltetem Gerät ist die Lampe heiß Schalten Sie den Reader aus und lassen Sie die Glühbirne mindestens 15 Minuten lang abkühlen bevor Sie versuchen sie auszutauschen Superficie molto calda Il gruppo lampada diventa molto caldo quando lo strumento è acceso Prima di tentare di sostituirlo spegne...

Page 186: ...nd reinstalled before repackaging the instrument for shipment Verzendingshardware Alle verzendingshardware moet worden verwijderd voordat het instrument wordt gebruikt en opnieuw worden geïnstalleerd voordat het instrument opnieuw wordt verpakt voor verzending Matériel d expédition Tout le matériel d expédition doit être retiré avant d utiliser l instrument et réinstallé avant de remballer l équip...

Page 187: ... deur niet om bij de filterkubus te komen als het instrument in bedrijf is Als u dit wel doet kan dit leiden tot ongeldige gegevens Cube de filtres modèles F Le tableau du cube de filtres internes du lecteur doit correspondre exactement au contenu du cube de filtres installé Utilisateurs de Gen5 Le tableau du cube de filtres du logiciel Gen5 doit correspondre exactement au contenu du cube de filtr...

Page 188: ... incernierato nella parte anteriore dello strumento Per evitare di produrre dati non validi non aprire lo sportello per accedere al filtro a cubo mentre lo strumento è in funzione Cubo de filtro modelos F La tabla de cubos de filtro interno del lector debe coincidir exactamente con el contenido del cubo de filtro instalado Usuarios de Gen5 la tabla de cubos de filtro del software Gen5 debe coincid...

Page 189: ...pouvez utiliser un stylet conçu pour les écrans tactiles résistifs Touchscreen Nutzen Sie Ihre Fingerspitze um den Touchscreen zu bedienen Verwenden Sie keinen spitzen Stylus oder Bleistift auf dem Touchscreen Dadurch wird die Oberfläche des Touchscreens beschädigt Sie können einen Stylus verwenden der für widerstandsgesteuerte Touchscreens entwickelt wurde Touchscreen Per utilizzare il touchscree...

Page 190: ... rayures Ne vaporisez jamais de solutions directement sur l écran tactile Touchscreen Verwenden Sie keine starken Lösungsmittel wie Alkohol Azeton Ammoniumchlorid Methylenchlorid und Kohlenwasserstoffe Diese Mittel verursachen bleibende Schäden am Touchscreen Faserstoffe wie Papiertücher können den Touchscreen zerkratzen In den Kratzern sammeln sich Schmutzpartikel und Reinigungsmittel Sprühen Sie...

Page 191: ...traîner l annulation de la garantie et potentiellement nuire à la performance de l instrument ou l endommager Ersatzteile Für die Wartung sollten nur genehmigte Ersatzteile verwendet werden Die Verwendung nicht genehmigter Ersatzteile und Zubehörkomponenten kann zum Verlust der Garantie führen und möglicherweise die Geräteleistung beeinträchtigen oder Schäden am Gerät verursachen Parti di ricambio...

Page 192: ...s doit être réservée au personnel technique qualifié Wartung Wartungsarbeiten an Komponenten im Geräteinneren sollten nur von qualifizierten Servicetechnikern durchgeführt werden Manutenzione Le procedure di manutenzione sui componenti interni devono essere eseguite esclusivamente da personale tecnico qualificato Revisión Solo puede realizar procedimientos de revisión de los componentes internos e...

Page 193: ...perator de bijsluiter van de test van de fabrikant volgen Het wordt beschouwd als een goede laboratoriumpraktijk om laboratoriummonsters te onderzoeken volgens de instructies en specifieke aanbevelingen die zijn opgenomen in de bijsluiter van de verpakking van de uit te voeren test Het niet uitvoeren van kwaliteitscontroles kan leiden tot foutieve testgegevens Contrôle de qualité du logiciel L opé...

Page 194: ...ura È considerata una buona pratica di laboratorio eseguire campioni di laboratorio in base alle istruzioni e alle raccomandazioni specifiche incluse nel foglietto della confezione del test relativo al test da condurre La mancata esecuzione delle verifiche di controllo qualità potrebbe dare luogo a dati di test errati Control de calidad del software El operador tiene que seguir las instrucciones d...

Page 195: ...sées par l opérateur Les caractéristiques de performance du logiciel de réduction des données n ont pas été établies par un test de diagnostic en laboratoire Les utilisateurs doivent évaluer l instrument et le logiciel pour PC conjointement à leur s test s spécifique s Cette évaluation doit comprendre la confirmation que les caractéristiques de performance pour le ou les tests spécifiques sont rem...

Page 196: ... de apparatuur niet wordt gebruikt volgens de richtlijnen en voorzorgsmaatregelen die in de gebruikersdocumentatie van het product staan vermeld kan dat leiden tot een gevaarlijke situatie Utilisation non spécifiée Ne pas utiliser l équipement conformément aux recommandations spécifiées dans la documentation utilisateur relative au produit peut entraîner des situations dangereuses Von den Vorschri...

Page 197: ...entraîner des erreurs de positionnement lors de l exécution du programme Die Verwendung anderer als in diesem Dokument beschriebener Laborgeräte kann zu Positionierungsfehlern bei der Programmausführung führen L uso di vetreria diversa da quella descritta in questo documento può causare errori di posizionamento durante l esecuzione del programma El uso de material de laboratorio diferente al descr...

Page 198: ...and qualification information for all models of the Synergy LX Document Revision History Part Number Revision Date Modifications 1501000N A December 2021 First issue Original Language EN Manufactured by Agilent Technologies Inc 5301 Stevens Creek Blvd Santa Clara CA 95051 ...

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