Synergy HTX Multi Mode Microplate Reader

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Product Information

Specifications:

• Product Name: Synergy HTX Multi-Mode Microplate Reader
• Manufacturer: BioTek Instruments, Inc.
• Release Date: July 2014
• Part Number: 1341000
• Revision: A

Product Usage Instructions:

1. Select an Appropriate Location

Choose a stable, flat surface with access to power outlets and
proper ventilation.

2. Connect the Power Supply

Plug in the power cord to the appropriate power source following
the manufacturer’s guidelines.

3. Unpack and Inspect the Dispense Module

Carefully unpack the dispense module and check for any visible
damage. Contact customer service if needed.

4. Install the Dispense Module

Follow the installation instructions provided in the manual to
correctly set up the dispense module.

FAQ:

Q: How can I get customer service or technical support?

A: You can contact BioTek Instruments, Inc. through their
website at www.biotek.com, phone at 888-451-5171 (toll-free in the
U.S.), or email at customercare@biotek.com.

Q: Where can I find service centers for repair?

A: BioTek has service centers worldwide. For technical
assistance, contact the Technical Assistance Center (TAC) at BioTek
World Headquarters US. You can also find service centers in
Germany, India, China, France, and Singapore. Contact the
respective centers for service or repair inquiries.

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Multi-Mode Microplate Reader
SynergyTM HTX
Operator’s Manual

Synergy HTX
Multi-Mode Microplate Reader Operator’s Guide
July 2014 2014 Part Number 1341000 Revision A BioTek Instruments, Inc.

ii | Preface
Notices
BioTek Instruments, Inc. Highland Park, P.O. Box 998 Winooski, Vermont 05404-0998 USA
All Rights Reserved
© 2014, BioTek® Instruments, Incorporated. No part of this publication may be reproduced, transcribed, or transmitted in any form, or by any means electronic or mechanical, including photocopying and recording, for any purpose other than the purchaser’s use without written permission of BioTek Instruments, Inc.
Trademarks
BioTek® is a registered trademark, and SynergyTM, Gen5TM, BioCellTM, 4-ZoneTM and BioStackTM are trademarks of BioTek Instruments, Inc. HartaTM is a trademark of Harta Instruments. Microsoft®, Windows®, and Excel® are either registered trademarks or trademarks of Microsoft Corporation in the United States and/or other countries. All other trademarks are the property of their respective holders.
Restrictions and Liabilities
Information in this document is subject to change and does not 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 BioTek or its affiliated dealers.
BioTek Instruments, Inc.

Contents

Contents | iii

Contact Information ………………………………………………….. v Customer Service and Sales…………………………………….. v Global Service and Support……………………………………… v
Document Conventions ……………………………………………… vii Revision History …………………………………………………….. viii Intended Use Statement ……………………………………………..ix Quality Control …………………………………………………………ix Warranty and Product Registration …………………………………ix Repackaging and Shipping ………………………………………….. x Warnings ……………………………………………………………….. x Hazards…………………………………………………………………. x Precautions ……………………………………………………………. xii CE Mark ………………………………………………………………. xiii
Directive 2004/108/EC: Electromagnetic Compatibility ….. xiii Directive 2006/95/EC Low Voltage (Safety)………………… xiii Directive 2002/96/EC: Waste Electrical and Electronic Equipment ……………………………………………………….. xiv Directive 98/79/EC: In Vitro Diagnostics (if labeled for this use) ……………………………………………………………….. xiv Electromagnetic Interference and Susceptibility ………………. xiv USA FCC CLASS A ………………………………………………. xiv Canadian Department of Communications Class A ………… xiv User Safety…………………………………………………………….xv Safety Symbols ……………………………………………………… xvi Introduction ………………………………………………………………. 1 Synergy HTX Multi-Mode Microplate Reader ……………………… 2 Package Contents …………………………………………………….. 3 Optional Accessories …………………………………………………. 4 Product Support & Service ………………………………………….. 5 Technical Assistance Center (TAC) …………………………….. 5 Returning Instruments for Service/Repair ……………………. 5 Applications Support ……………………………………………… 5 Installation………………………………………………………………… 7 Product Registration ………………………………………………….. 8 1: Unpack and Inspect the Reader ………………………………… 8 2: Remove the Shipping Panel ……………………………………. 10 3: Remove the Microplate Carrier Shipping Screw…………….. 11 4: Install the Fluorescence Lamp Assembly…………………….. 12
Synergy HTX Operator’s Manual

iv | Preface
5: Select an Appropriate Location………………………………… 13 6: Connect the Power Supply……………………………………… 14 7: Unpack and Inspect the Dispense Module …………………… 15 8: Install the Dispense Module……………………………………. 18
Record Syringe Calibration Values……………………………. 18 Install the Dispenser ……………………………………………. 18 9: Connect the Host Computer …………………………………… 20 10: Install Gen5 Software …………………………………………. 20 11: Turn on the Reader ……………………………………………. 20 12: Establish Communication …………………………………….. 21 13: Set the Dispenser Calibration Values ……………………….. 22 14: Run a System Test …………………………………………….. 23 15: Test the Injector System ……………………………………… 24 Operational/Performance Qualification ………………………….. 26 Repackaging and Shipping Instructions …………………………. 26 Getting Started…………………………………………………………. 31 Key Components ……………………………………………………. 32 Power Switch, Carrier Eject Button, Microplate Carrier …… 32 Lamp Assembly and Filter Wheel Access ……………………. 33 Excitation and Emission Filter Wheels ……………………….. 34 Installing the Time- Resolved Fluorescence Cartridge …….. 37 Configuring the System for Luminescence Measurements .. 38 The External Dispense Module ………………………………… 39 Gen5 Software ………………………………………………………. 41 Viewing/Updating the Filter and Wavelengths Tables …….. 42 Creating Protocols and Experiments …………………………. 43 Controlling the Dispense Module ……………………………… 45 Recommendations for Optimum Performance………………….. 48 Incubation and Partial Plates ………………………………….. 49 Preventive Maintenance …………………………………………….. 51 Recommended Maintenance Schedule…………………………… 52 Overview …………………………………………………………. 52 Daily Cleaning for the Dispense Module …………………….. 52 Recommended Maintenance Schedule ………………………. 53 Warnings and Precautions …………………………………………. 54 Cleaning Exposed Surfaces………………………………………… 55 Inspect/Clean Excitation and Emission Filters ………………….. 56 Flush/Purge the Fluid Path ………………………………………… 57 Running a Dispense Protocol (Optional) ………………………… 58 Empty/Clean the Tip Priming Trough ……………………………. 59 Clean the Priming Plate…………………………………………….. 59 Clean the Internal Components…………………………………… 60
BioTek Instruments, Inc.

Contents | v
Required Materials………………………………………………. 61 Removing the Reader’s Shroud……………………………….. 62 Removing the Internal Tubes and Injector Heads …………. 64 Cleaning the Internal Tubes and Injector Heads …………… 67 Cleaning the Optical Probes …………………………………… 68 Cleaning the Reader’s Internal Surface ……………………… 76 Reassembling the Components……………………………….. 77 Performance Check……………………………………………… 79 As-Needed Maintenance …………………………………………….. 81 Purpose ……………………………………………………………….. 82 Required Materials ………………………………………………….. 83 Procedure for Models without Injectors …………………………. 84 Routine Procedure for Models with Injectors …………………… 85 Clean Exposed Surfaces ……………………………………….. 85 Decontaminate the Fluid Lines………………………………… 86 Rinse the Fluid Lines ……………………………………………. 87 Clean the Internal Tubing and Injector Heads ……………… 87 Clean the Tip Priming Trough and Priming Plate …………… 88 Alternate Procedure for Models with Injectors …………………. 89 Instrument Qualification ……………………………………………. 91 Overview ……………………………………………………………… 92 IQ/OQ/PQ …………………………………………………………….. 92 Recommended Qualification Schedule …………………………… 94 System Test………………………………………………………….. 95 Absorbance Plate Test ……………………………………………..100 Test Plate Certificates ………………………………………….100 Define Absorbance Test Plate Parameters ………………….100 Run the Absorbance Plate Test ……………………………….101 Results and Troubleshooting Tips…………………………….104 Luminescence Tests ………………………………………………..105 Harta Plate Test …………………………………………………106 Gen5 Protocol Reading Parameters ………………………….107 Troubleshooting …………………………………………………109 Absorbance Liquid Tests …………………………………………..109 Absorbance Liquid Test 1………………………………………110 Absorbance Liquid Test 2………………………………………112 Absorbance Liquid Test 3 (optional) …………………………114 Fluorescence Tests………………………………………………….116 Required Materials………………………………………………117 Test Solutions……………………………………………………118 Procedure ………………………………………………………..119 Results Analysis …………………………………………………119
Synergy HTX Operator’s Manual

vi | Preface
Troubleshooting …………………………………………………120 Pipette Map ………………………………………………………121 Gen5 Protocol Reading Parameters ………………………….122 Fluorescence Tests Using Methylumbelliferone …………….126 Dispense Module Tests …………………………………………….131 Required Materials………………………………………………132 Alternate Test Solutions ……………………………………….133 Procedure for Models with Absorbance Capabilities ……….133 Procedure for Models without Absorbance Capabilities …..135 Results Analysis …………………………………………………136 Gen5 Test Protocols for Models with Absorbance Capabilities137 Gen5 Test Protocols for Models without Absorbance Capabilities ………………………………………………………139 Create the Dispense Protocols ………………………………..139 Specifications …………………………………………………………. 143 General Specifications ……………………………………………..144 Absorbance Specifications …………………………………………145 Fluorescence Specifications ……………………………………….147 Luminescence Specifications………………………………………148 Models with Injectors ………………………………………………149 Error Codes ……………………………………………………………. 151 Error Codes Overview………………………………………………152 Contact Info: BioTek Service/TAC ……………………………152 Error Codes ………………………………………………………….153
BioTek Instruments, Inc.

Contact Information | v

Contact Information

Customer Service and Sales

Internet: Phone:
Fax: Email:

www.biotek.com 888-451-5171 (toll-free in the U.S.) 802-655-4740 (outside the U.S.) 802-655-7941 customercare@biotek.com

Global Service and Support

BioTek instrument service and repair is available worldwide at one of BioTek’s International Service Centers and in the field at your location. For technical assistance, contact the Technical Assistance Center (TAC) at BioTek World Headquarters US. To arrange for service or repair of your instrument, contact the office nearest you.

BioTek World Headquarters US
Mailing Address: PO Box 998, Highland Park Winooski, VT 05404-0998 United States Service Shipping Address: 15 Tigan Street Winooski, VT 05404 Phone: 800-242-4685 Outside US: 802-655-4740 Fax: 802-654-0638
Email: TAC@biotek.com Website: www.biotek.com
BioTek Germany Service Center & European Coordination Center
Kocherwaldstrasse 34 D-74177 Bad Friedrichshall Germany Phone: +49 (0) 71369680 Fax: +49 (0) 7136968111 Email: info@biotek.de Website: www.biotek.de
BioTek India Unit 223, Linkway Estate New Link Road, Malad West Mumbai 400064

BioTek China Room 304, Tower D Ocean International Center 62 Middle 4th East Ring Road Chaoyang District Beijing 100025 P.R. China
Phone: +86 (10) 85865569 Fax: +86 (10) 85861829
Email: infochina@biotek.com Website: www.biotekchina.com.cn
BioTek Instruments SAS Bureau de liaison France 50 avenue d’Alsace 68025 Colmar Cedex France Phone: +33 (3) 89206329 Fax: +33 (3) 89204379 Email: info@biotek.fr Website: www.biotek.fr
BioTek Singapore 20 Science Park Road #01-08A Teletech Park Singapore 117674

Synergy HTX Operator’s Manual

vi | Preface

India Phone: +91 (22) 28789966 Fax: +91 (22) 28759944 Email: biotek@biotek.in Website: www.biotek.in

Phone: +65 65922100 Fax: +65 67772611 Email: singapore@biotek.com Website: www.biotek.com

BioTek South Korea
3F, Gyungnam building, 830-48 Yeoksam-dong, Gangnam-gu Seoul, South Korea (135-936) Phone: +82 (0) 2-562-4740 Fax: +82 (0) 2-562-4750 Email: korea@biotek.com Website: www.biotekinstruments.co.kr

BioTek Switzerland
Zentrum Fanhöfli 8 6014 Luzern Switzerland Phone: +41 (41) 2504060 Fax: +41 (41) 2505064 Email: info@biotek.ch Website: www.biotek.ch

BioTek United Kingdom (UK)
6 Bull Street Potton, Bedfordshire SG19 2NR United Kingdom Phone: +44 (1767) 262000 Fax: +44 (1767) 262330 Email: info@biotek.uk.com Website: www.biotek.uk.com

BioTek Instruments, Inc.

Document Conventions | vii

Document Conventions

This icon calls attention to important safety notes.

Warning! Caution
Note: italic

A Warning indicates the potential for bodily harm and tells you how to avoid the problem.
A Caution indicates potential damage to the instrument and tells you how to avoid the problem.
Bold text is primarily used for emphasis.
Topics that apply only to specific Synergy HTX models are preceded by a notice in italics, for example: Applies only to Synergy HTX models with injectors.
This icon calls attention to important information.

Synergy HTX Operator’s Manual

viii | Preface

Revision History

Rev Date A 7/2014

Changes Initial release

BioTek Instruments, Inc.

Intended Use Statement | ix
Intended Use Statement
· The Synergy HTX is a single-channel absorbance, fluorescence, and luminescence microplate reader that uses a dual-optics design to perform measurements of samples in a microplate format. The performance characteristics of the data reduction software have not been established with any laboratory diagnostic assay. The user must evaluate this instrument and Gen5 software in conjunction with their specific assay(s). This evaluation must include the confirmation that performance characteristics for the specific assay(s) are met.
· If the instrument has an “IVD” label, it may be used for clinical and non- clinical purposes, including research and development. If there is no such label, the instrument may be used only for research and development or other non-clinical purposes.
Quality Control
It is considered good laboratory practice to run laboratory 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
Take a moment to review the Warranty information that shipped with your product. Please also register your product with BioTek to ensure that you receive important information and updates about the product(s) you have purchased. You can register online through the Customer Resource Center at www.biotek.com or by calling 888/451-5171 or 802/655-4740.
Synergy HTX Operator’s Manual

x | Preface

Repackaging and Shipping

If you need to ship the instrument to BioTek for service or repair, contact BioTek for a service authorization number, and be sure to use the original packing materials. Other forms of commercially available packaging are not recommended and can void the warranty. If the original packing materials have been damaged or lost, contact BioTek for replacement packing.

Warnings
Operate the instrument on a level, stable surface away from excessive humidity. Bright sunlight or strong incandescent light can reduce the linear performance range of the instrument. Measurement values may be affected by extraneous particles (such as dust) in the microplate wells. A clean work area is necessary to ensure accurate readings. When operated in a safe environment according to the instructions in this document, there are no known hazards associated with the instrument. However, the operator should be aware of certain situations that could result in serious injury; these may vary depending on the instrument model. See Hazards and Precautions.
Hazards
The following hazard warnings are provided to help avoid injury:
Warning! Internal Voltage. Always turn off the power switch and unplug the power supply before cleaning the outer surface of the instrument or removing its top case. Warning! Power Rating. The instrument’s power supply or power cord must be connected to a power receptacle that provides voltage and current within the specified rating for the system. Use of an incompatible power receptacle may produce electrical shock and fire hazards. Warning! Electrical Grounding. Never use a plug adapter to connect primary power to the external power supply. Use of an adapter disconnects the utility ground, creating a severe shock hazard. Always connect the power cord directly to an appropriate receptacle with a functional ground.

BioTek Instruments, Inc.

Hazards | xi
Warning! Service. Only qualified technical personnel should perform service procedures on internal components. Warning! Accessories. Only accessories that meet the manufacturer’s specifications shall be used with the instrument. Warning! The instrument weighs approximately 38 pounds (17 kg). Use two people when lifting and carrying the instrument. Warning! Lubricants. Do not apply lubricants to the microplate carrier or carrier track. Lubricant on the carrier mechanism or components in the carrier compartment will attract dust and other particles, which may obstruct the carrier path and cause the instrument to produce an error. Warning! Liquids. Avoid spilling liquids on the instrument; fluid seepage into internal components creates a potential for shock hazard or instrument damage. If a spill occurs while a program is running, abort the program and turn the instrument off. Wipe up all spills immediately. Do not operate the instrument if internal components have been exposed to fluid. Warning! Unspecified Use. Failure to operate this equipment according to the guidelines and safeguards specified in this manual could result in a hazardous condition. Warning! Software Quality Control. The operator must follow the manufacturer’s assay package insert when modifying software parameters and establishing reading, washing, or dispensing methods. Failure to conduct quality control checks could result in erroneous test data.
Warning! Reader Data Reduction Protocol. No limits are applied to the raw absorbance data. All information exported via computer control must be thoroughly analyzed by the operator. Warning! Potential Biohazards. Some assays or specimens may pose a biohazard. Adequate safety precautions should be taken as outlined in the assay’s package insert. This hazard is noted by the symbol shown here. Always wear safety glasses and appropriate protective equipment, such as chemically resistant rubber gloves and apron. Warning! Hot Surface. The lamp assembly is hot when the instrument is turned on. Turn off the reader and allow the lamp to cool down before attempting replacement. Warning! Pinch Hazard. Some areas of the dispense module can present pinch hazards when the instrument is operating. The module is marked with one of the symbols shown here. Keep hands/fingers clear of these areas when the instrument is operating.
Synergy HTX Operator’s Manual

xii | Preface
Precautions
The following precautions are provided to help avoid damage to the instrument:
Caution: Service. The instrument should be serviced by BioTek authorized service personnel. Only qualified technical personnel should perform troubleshooting and service procedures on internal components. Caution: Spare Parts. Only approved spare parts should be used for maintenance. The use of unapproved spare parts and accessories may result in a loss of warranty and potentially impair instrument performance or cause damage to the instrument. Caution: Environmental Conditions. Do not expose the instrument to temperature extremes. For proper operation, ambient temperatures should remain within the range listed in the Specifications section. Performance may be adversely affected if temperatures fluctuate above or below this range. Storage temperature limits are broader. Caution: Sodium Hypochlorite. Do not expose any part of the instrument to the recommended diluted sodium 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: Disposal. Dispose of the instrument according to Directive 2002/96/EC, “on waste electrical and electronic equipment (WEEE),” or local ordinances. Caution: Warranty. Failure to follow preventive maintenance protocols may void the warranty. Caution: Shipping Hardware. All shipping hardware must be removed before operating the instrument and reinstalled before repackaging the instrument for shipment. Caution: Electromagnetic Environment. Per IEC 61326-2-6 it is the user’s responsibility to ensure that a compatible electromagnetic environment for this instrument is provided and maintained in order that the device will perform as intended. Caution: Electromagnetic Compatibility. Do not use this device in close proximity to sources of strong electromagnetic radiation (e.g., unshielded intentional RF sources), because these may interfere with the proper operation.
BioTek Instruments, Inc.

CE Mark | xiii
CE Mark
Based on the testing described below and information contained herein, this instrument bears the CE mark
See the Declaration of Conformity for more information.
Directive 2004/108/EC: Electromagnetic Compatibility
Emissions–Class A
The system has been type-tested by an independent, accredited testing laboratory and found to meet the requirements of EN 61326-1: Class A for Radiated Emissions and Line Conducted Emissions. Verification of compliance was conducted to the limits and methods of EN 55011 ­ (CISPR 11) Class A. In a domestic environment it may cause radio interference, in which case you may need to mitigate the interference.
Immunity
The system has been type-tested by an independent, accredited testing laboratory and found to meet the requirements of EN 61326-1 and EN 61326-2-6 for Immunity. Verification of compliance was conducted to the limits and methods of the following:
EN 61000-4-2, Electrostatic Discharge EN 61000-4-3, Radiated EM Fields EN 61000-4-4, Electrical Fast Transient/Burst EN 61000-4-5, Surge Immunity EN 61000-4-6, Conducted Disturbances from RFI EN 61000-4-11, Voltage Dips, Short Interruptions and Variations
Directive 2006/95/EC Low Voltage (Safety)
The system has been type-tested by an independent testing laboratory and was found to meet the requirements of this Directive. Verification of compliance was conducted to the limits and methods of the following: EN 61010-1. “Safety requirement for electrical equipment for measurement, control and laboratory use. Part 1, General requirements.” EN 61010-2-081, “Particular requirements for automatic and semi-automatic laboratory equipment for analysis and other purposes.”
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xiv | Preface
EN 61010-2-010, “Particular requirements for laboratory equipment for the heating of materials.”
Directive 2002/96/EC: Waste Electrical and Electronic Equipment
Disposal Notice: Dispose of the instrument according to Directive 2002/96/EC, “on waste electrical and electronic equipment (WEEE)” or local ordinances.
Directive 98/79/EC: In Vitro Diagnostics (if labeled for this use)
· Product registration with competent authorities. · Traceability to the U.S. National Institute of Standards and Technology (NIST). · EN 61010-2-101, “Particular requirements for in vitro diagnostic (IVD) medical
equipment.”
Electromagnetic Interference and Susceptibility
USA FCC CLASS A
RADIO AND TELEVISION INTERFERENCE Note: This equipment has been tested and found to comply with the limits for a Class A digital device, pursuant to Part 15 of the FCC Rules. These limits are designed to provide reasonable protection against harmful interference when the equipment is operated in a commercial environment. Like all similar equipment, this equipment generates, uses, and can radiate radio frequency energy and, if not installed and used in accordance with the instruction manual, may cause harmful interference to radio communications. Operation of this equipment in a residential area is likely to cause interference, in which case users will be required to correct the interference at their own expense. In order to maintain compliance with FCC regulations, shielded cables must be used with this equipment. Operation with non-approved equipment or unshielded cables is likely to result in interference to radio and television reception.
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’emet pas de bruits radioelectriques depassant les limites applicables aux appareils numerique de la Class A prescrites dans le Reglement sur le brouillage radioelectrique edicte par le ministere des Communications du Canada.
BioTek Instruments, Inc.

User Safety | xv
User Safety
This device has been type-tested by an independent laboratory and found to meet the requirements of the following:
· Underwriters Laboratories UL 61010-1, “Safety requirements for electrical equipment for measurement, control and laboratory use; Part 1: General requirements.”
· Canadian Standards Association CAN/CSA C22.2 No. 61010-1, “Safety requirements for electrical equipment for measurement, control and laboratory use; Part 1: General requirements.”
· EN 61010 Standards, see CE Mark starting on page xiii.
Synergy HTX Operator’s Manual

xvi | Preface

Safety Symbols

Some of these symbols appear on the instrument or accessories:

Alternating current Courant alternatif Wechselstrom Corriente alterna Corrente alternata
Direct current Courant continu Gleichstrom Corriente continua Corrente continua
On (Supply) Marche (alimentation) Ein (Verbindung mit dem Netz) Conectado Chiuso
Off (Supply) Arrêt (alimentation) Aus (Trennung vom Netz) Desconectado Aperto (sconnessione dalla rete di alimentazione)
Warning, risk of electric shock Attention, risque de choc électrique Gefährliche elektrische schlag Precaución, riesgo de sacudida eléctrica Attenzione, rischio di scossa elettrica
Warning, hot surface Attention, surface chaude Warnen, heiße Oberfläche Precaución, superficie caliente Attenzione, superficie calda

Both direct and alternating current Courant continu et 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
Caution (refer to accompanying documents) Attention (voir documents d’accompanement) Achtung siehe Begleitpapiere Atención (vease los documentos incluidos) Attenzione, consultare la doc annessa
Warning, risk of crushing or pinching Attention, risque d’écrasement et pincement Warnen, Gefahr des Zerquetschens und Klemmen Precaución, riesgo del machacamiento y sejeción Attenzione, rischio di schiacciare ed intrappolarsi
Warning, potential biohazards Attention, risques biologiques potentiels Warnung! Moegliche biologische Giftstoffe Atención, riesgos biológicos Attenzione, rischio biologico

BioTek Instruments, Inc.

In vitro diagnostic medical device Dispositif médical de diagnostic in vitro Medizinisches In-VitroDiagnostikum Dispositivo médico de diagnóstico in vitro Dispositivo medico diagnostico in vitro
Consult instructions for use Consulter la notice d’emploi Gebrauchsanweisung beachten Consultar las instrucciones de uso Consultare le istruzioni per uso

Safety Symbols | xvii
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 Raccolta separata delle apparecchiature elettriche ed elettroniche

Synergy HTX Operator’s Manual

xviii | Preface
BioTek Instruments, Inc.

Chapter 1
Introduction
This chapter introduces the Synergy HTX, describes its key features, lists its package contents, and provides contact information for technical assistance. Synergy HTX Multi-Mode Microplate Reader …………………………. 2 Package Contents…………………………………………………………. 3 Optional Accessories ……………………………………………………… 4 Product Support & Service ………………………………………………. 5
Technical Assistance Center (TAC)…………………………………. 5 Returning Instruments for Service/Repair………………………… 5 Applications Support …………………………………………………. 5

2 | Chapter 1: Introduction
Synergy HTX Multi-Mode Microplate Reader
The Synergy HTX is a single-channel microplate reader available with absorbance, fluorescence, and luminescence detection. It is computer- controlled using BioTek’s Gen5 software for all operations including data reduction and analysis. Synergy HTX is robot accessible and compatible with BioTek’s BioStack Microplate Stacker. When making fluorescence determinations, the Synergy HTX uses a tungsten quartz halogen lamp with interference filters for wavelength specificity in conjunction with a photomultiplier (PMT) tube detector. The Synergy HTX has both top and bottom probes for fluorescence measurements. The top probe can be adjusted vertically for the correct reading height, via Gen5’s Read Height reading parameter (see Chapter 3, Getting Started). Luminescence is measured by the low-noise PMT detector through an empty filter position in the Emission filter wheel. A filter can also be left in place if light filtering is necessary. Absorbance measurements are made by switching to a xenon 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 HTX has a 4-Zone temperature control from 4°C over ambient to 50°C, controlled via a software-adjustable gradient. Internal plate shaking, with both linear and orbital modes, is supported to ensure that reagents are properly mixed prior to reading. Both Synergy HTX models support the reading of 6-, 12-, 24-, 48-, 96-, and 384-well microplates with standard 128 x 86 mm geometry, as well as the BioTek Take3 and Take3 Trio Micro-Volume Plates. Absorbance mode reads plates up to 0.8″ (20.3 mm) in height; fluorescence mode reads plates up to 1.25″ (31.75 mm). Polymerase Chain Reaction (PCR) tubes up to 1.25″ (31.75 mm) are also readable with the use of existing adapter plates. For models with time-resolved fluorescence (TRF) capability, the TRF option allows measurements by using the xenon flash light source in conjunction with the PMT measurement detector. A special cartridge installed in the Excitation filter wheel location is required. Models with injectors support dual-reagent dispensing to 6-, 12-, 24-, 48-, 96-, and 384-well microplates with standard 128 x 86 mm geometry. An external dispense module pumps fluid from the supply bottles to the two injectors located inside the instrument. Both injectors are positioned directly above the bottom probe, and fluid is injected into one well at a time.
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Package Contents | 3

Package Contents

Part numbers and package contents are subject to change. Contact BioTek Customer Care with any questions.

Item Synergy HTX Operator’s Manual
Power supply
Power cord set (specific to installation environment): Europe (Schuko) USA/International United Kingdom Australia/New Zealand
RS-232 serial cable
USB cable with USB Driver Software
Wrench Fluorescence lamp assembly (Note: The replacement lamp assembly is PN 7080500) Filter “plugs” (2) (also referred to as “dummy filters” or “blanks”)
Plastic storage bag and fastener strips
Time-Resolved Fluorescence cartridge assembly (“T” models only)
Models with injectors, an external dispense module (packed separately), with the following accessories:
Outlet tubes (2, plus 2 spare) from dispense module to instrument Inlet tubes (2) from supply bottles to syringe drives 250 µl syringes (2) Syringe thumbscrews (2) Priming plate Injector tip priming trough Dispense module communication cable Dispense module front cover Supply bottles (2, 30 mL) Supply bottle holder assemblies (2) Injector tip cleaning stylus and plastic storage bag

• If applicable to your reader model.

Part # 1341000
76061
75010 75011 75012 75013 75034 75108 7090204 7772028 7080501
7082073 —
7090523
7082120 7082121 7083000
19511 7132158 1342017
75107 7082137 7122609 7090564 2872304

Synergy HTX Operator’s Manual

4 | Chapter 1: Introduction

Optional Accessories

Accessory availability and part numbers are subject to change. Contact BioTek Customer Care with questions or visit www.biotek.com and use the Accessories search tool.

Item 7-filter Absorbance Test Plate Fluorescence Test Plate Product Qualification (IQ-OQ-PQ) package
PCR Tube Adapter Plates

Part # 7260522
7092092
1340508
6002072 and 6002076

Terasaki Adapter Plate Take3 Micro-Volume Plate Take3 Trio Micro-Volume Plate BioCell Quartz Vessel and Adapter Plate

7330531 TAKE3
Take3Trio 7272051/7270512

Additional Fluorescence Filters; contact BioTek for part numbers and availability

The Synergy HTX is compatible with the BioStack Microplate Stacker. Contact BioTek or visit our website to learn more.

For Use with Liquid Tests (see Chapter 5)
Absorbance Liquid Test Solutions: BioTek Wetting Agent Solution BioTek QC Check Solution #1 25 mL 125 mL
Dispense Module Liquid Test Solution: BioTek Green Test Dye BioTek Blue Test Dye BioTek QC (Yellow) Test Dye
Individual Fluorescence Liquid Test Solutions: Sodium Fluorescein Powder Liquid Test Kit using Sodium Fluorescein Liquid Test Kit using Methylumbelliferone (“MUB”)

Part #
7773002
7120779 7120782
7773003 7773001 7120782
98155 7160013 7160012

BioTek Instruments, Inc.

Product Support & Service | 5

Product Support & Service

Technical Assistance Center (TAC)

If your instrument(s) or software fails to function properly, if you have questions about how to use or maintain our products, or if you need to send an instrument to BioTek for service or repair, please contact our Technical Assistance Center. BioTek’s “TAC” is open from 8:30 AM to 5:30 PM (EST), Monday through Friday, excluding standard U.S. holidays. You can send a fax or an e-mail any time. You can also request technical assistance via our website: www.biotek.com.

Phone: 800-242-4685 or 802-655-4740

Fax: 802-654-0638

E-Mail: tac@biotek.com Web: www.biotek.com

Please be prepared to provide the following information:

· Your name and company information, along with a daytime phone or fax number, and/or an e-mail address

· The product name, model, and serial number

· The onboard software part number and version (available through Gen5 at System > Instrument Configuration > Get Basecode Information)

· Gen5 software version information (Help > About Gen5)

· For troubleshooting assistance or instruments needing repair, the specific steps that produce your problem and any error codes displayed in Gen5 (see also Appendix C, Error Codes)

· A text file of the diagnostic history of the instrument (available via Gen5 by selecting System > Diagnostics > History, then selecting the appropriate file and clicking Export)

Returning Instruments for Service/Repair

If you need to return an instrument to BioTek for service or repair, please contact the TAC for a service authorization number before shipping the instrument. Repackage the instrument properly (see Chapter 2, Installation), write the number on the shipping box, and ship to BioTek.

Applications Support

BioTek’s fully equipped Application Laboratory provides our on-staff scientists with the means to assist you with the integration of our instrumentation and software with your unique scientific applications. If you are having difficulty with optimizing fluorescence sensitivity or integrating a unique data reduction transformation, or you are just looking for a recommendation on an appropriate fluorophore, contact us.

Phone: 888-451-5171

E-Mail: applications@biotek.com

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6 | Chapter 1: Introduction
BioTek Instruments, Inc.

Chapter 2
Installation
This chapter includes instructions for unpacking and setting up the Synergy HTX and, if applicable, the external dispense module. Instructions are also included for repackaging the reader and dispense module for shipment.
Product Registration ……………………………………………………… 8 1: Unpack and Inspect the Reader …………………………………….. 8 2: Remove the Shipping Panel ……………………………………….. 10 3: Remove the Microplate Carrier Shipping Screw ………………… 11 4: Install the Fluorescence Lamp Assembly ………………………… 12 5: Select an Appropriate Location ……………………………………. 13 6: Connect the Power Supply …………………………………………. 14 7: Unpack and Inspect the Dispense Module ………………………. 15 8: Install the Dispense Module ……………………………………….. 18 9: Connect the Host Computer ……………………………………….. 20 10: Install Gen5 Software……………………………………………… 20 11: Turn on the Reader ………………………………………………… 20 12: Establish Communication …………………………………………. 21 13: Set the Dispenser Calibration Values …………………………… 22 14: Run a System Test ………………………………………………… 23 15: Test the Injector System …………………………………………. 24 Operational/Performance Qualification………………………………. 26 Repackaging and Shipping Instructions …………………………….. 26

8 | Chapter 2: Installation
Product Registration
Please register your product with BioTek to ensure that you receive important information and updates about the products you have purchased. Contact the Customer Resource Center (CRC) at www.biotek.com or by calling 888-451-5171 or 802-655-4740.

1: Unpack and Inspect the Reader

Important! Save all packaging materials. If you need to ship the reader to BioTek for repair or replacement, you must use the original materials. Using other forms of commercially available packaging, or failing to follow the repackaging instructions, may void your warranty. Improper packaging that results in damage to the instrument may lead to additional charges.
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. Keep the shipping boxes and the packaging materials for the carrier’s inspection. BioTek will arrange for repair or replacement of your reader immediately.

1. Open the outer shipping box. Remove the foam blocks to access the inner box. 2. Carefully open the inner shipping box. Remove the accessories box and set it aside.
Remove the vertical supports.
Warning! The instrument weighs approximately 38 pounds (17 kg). Use two people when lifting and carrying the instrument.
3. The Synergy HTX is attached to a shipping panel that has two handles for lifting. Locate 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.

BioTek Instruments, Inc.

1: Unpack and Inspect the Reader | 9
accessories box
Figure 1: Unpacking the reader Synergy HTX Operator’s Manual

10 | Chapter 2: Installation
2: Remove the Shipping Panel
1. Carefully tip the reader onto its back. 2. Using a screwdriver, remove the four screws and washers attaching the shipping
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 fastener strips to attach the pocket to the back of the reader for storage. Do not block any air vents. See Figure 2 on the next page. 5. Place the panel back into the inner shipping box for storage.

Important: Reattach the shipping panel before repackaging the Synergy HTX for shipment.

BioTek Instruments, Inc.

3: Remove the Microplate Carrier Shipping Bolt | 11

Figure 2: Removing the shipping panel

3: Remove the Microplate Carrier Shipping Bolt

Important: Remove the microplate carrier shipping bolt before turning on the Synergy HTX.

1. Pull down the microplate loading door on the front of the reader.
2. Using the supplied wrench, remove the carrier shipping bolt with its o-ring and warning tag.

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12 | Chapter 2: Installation
3. Store the wrench, bolt, o-ring, and tag in a safe place, in case you need to ship the instrument back to BioTek.

shipping bolt o-ring

shipping bolt warning tag

Figure 3: Removing the microplate carrier shipping bolt

Important: Replace the microplate carrier shipping bolt before repackaging the Synergy HTX for shipment. Please contact BioTek if you have misplaced the bolt (PN 1342008) and/or its o-ring (PN 49259).

4: Install the Fluorescence Lamp Assembly

Applies only to Synergy HTX models with fluorescence capability.

Important: Do not touch the glass lenses! Fingerprints on the condenser lens or heat absorber may negatively affect performance.

BioTek Instruments, Inc.

5: Select an Appropriate Location | 13
Warning! The fluorescence lamp assembly is hot when the instrument is powered on. If the instrument is on, turn it off and allow the lamp to cool down before attempting to replace it.
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 the way into the compartment.
4. Plug the lamp cables into the power source located to the right of the lamp. Either cable can be plugged into either socket.
5. Close the hinged door.
Figure 4: Installing the fluorescence lamp assembly (replacement lamp PN 7080500)
5: Select an Appropriate Location
Install the Synergy HTX on a level surface in an area where ambient temperatures between 18ºC (64ºF) and 40ºC (104ºF) can be maintained. The reader is sensitive to extreme environmental conditions. Avoid the following:
Synergy HTX Operator’s Manual

14 | Chapter 2: Installation
· Excessive humidity: Condensation directly on the sensitive electronic circuits can cause the reader to fail internal self-checks. The specified relative 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. If you will be installing BioTek’s BioStack Microplate Stacker for operation with the Synergy HTX, you may wish to seat the BioStack and the reader in their aligning plates at this time. Refer to the BioStack Operator’s Manual for more information.
6: Connect the Power Supply
Warning! Power Rating. The power supply must be connected to a power receptacle that provides voltage and current within the specified rating for the system. Use of an incompatible power receptacle may produce electrical shock and fire hazards. Warning! Electrical Grounding. Never use a plug adapter to connect primary power to the external power supply. Use of an adapter disconnects the utility ground, creating a severe shock hazard. Always connect the power cord directly to an appropriate receptacle with a functional ground.
1. Plug the rounded end of the power supply’s cord into the power inlet on the rear of the reader.
2. Connect the power cord to the external power supply. 3. Plug the power cord into an appropriate power receptacle.
BioTek Instruments, Inc.

7: Unpack and Inspect the Dispense Module | 15

7: Unpack and Inspect the Dispense Module
Applies only to Synergy HTX models with injectors.

Important! Save all packaging materials. If you need to ship the dispense module to BioTek for repair or replacement, you must use the original materials. Using other forms of commercially available packaging, or failing to follow the repackaging instructions, may void your warranty.
During the unpacking process, 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.

1. Open the outer shipping box. Remove the foam cap, inner shipping box, and accessories box.

Figure 5: Unpacking the dispense module’s outer shipping box Synergy HTX Operator’s Manual

16 | Chapter 2: Installation

2. Using no sharp tools, open the box containing the dispense module. Remove the two reagent bottle holders and the cardboard shipping insert. Lift out the module and place it on a level surface.

Reagent bottle holders
(2)

SChairpdpbinogairndseinrtsert

DiDspisepnesne se momduolde ule

Inner Innersshhipipppinigngbox
box
Figure 6: Unpacking the dispense module’s inner shipping box
3. Open the accessories box. Remove and identify its contents (see Figure 7 on the next page): · 2 inlet tubes, packaged in plastic cylinders · 4 outlet tubes, packaged in plastic bags · 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 fastener strips · 2 metal thumbscrews · 1 stylus (wire) packaged in a small plastic cylinder · 1 dispense module cover · 1 dispense module cable
BioTek Instruments, Inc.

Top foam end cap
Inlet tubes (2) Outlet tubes (4)

7: Unpack and Inspect the Dispense Module | 17
Dispense module cable
Dispense module cover
Syringes (2)

Bottom foam end cap (with
cutouts)
Accessories box
Figure 7: Unpacking the dispense module’s accessories

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18 | Chapter 2: Installation
8: Install the Dispense Module
Applies only to Synergy HTX models with injectors.
Record Syringe Calibration Values
If applicable, perform these steps to record calibration information and then install the dispenser.
1. Record the syringe calibration values that were set at BioTek: a. On the back panel of the dispenser box, locate the two labels that show the six target volumes (200, 80, 40, 20, 10, and 5) and the six corresponding calibration values. b. Record the 12 calibration values in the IQ Checklist.
Install the Dispenser
1. Place the module to the left side or on top of the reader. 2. On the rear panel of the reader, identify the SYRINGE 1 and SYRINGE 2 tubing
ports. Remove the nylon screws from both ports and set them aside. 3. Open two of the plastic bag containing the injector tubes and tips. Remove the
clear plastic shrouds from the tubes. Put the other two bags in a safe place; they are spares. 4. Remove the two inlet tubes from their plastic canisters. 5. Identify the two syringe valves on the dispense module. Each is labeled with a left-pointing arrow.
When installing the inlet and outlet tubes, do not use any tools. Finger- tighten only!
6. Screw the fitting of one inlet tube into the right side of the Syringe 1 valve. 7. Screw one end of one outlet tube into the left side of the Syringe 1 valve. 8. Screw the other end of the outlet tube into the SYRINGE 1 port on the rear of
the reader. 9. Repeat these steps to attach the inlet and outlet tubing for Syringe 2.
It is critical that the tubing is installed in the correct ports. Otherwise, injected fluid may miss the intended well.
10. Remove the two syringes from their protective boxes. They are identical and interchangeable. Each syringe should already be assembled in one piece, but if for some reason there are two separate pieces, assemble them now: Insert the
BioTek Instruments, Inc.

8: Install the Dispense Module | 19 white tip of the syringe plunger into the barrel of the syringe, and gently push it all the way into the barrel. 11. Install both syringes: a. Hold the syringe vertically with the threaded end at the top. b. Screw the top of the syringe into the bottom of the syringe valve. Finger-
tighten only. c. Carefully pull down the bottom of the syringe until it rests inside the hole
in the bracket. d. Pass a thumbscrew up through this hole, and thread it into the bottom of
the syringe. Hold the syringe to prevent it from rotating while tightening the thumbscrew. Finger-tighten only.
12. Locate the dispenser cable. Plug one end into the port on the left side of the dispenser. Plug the other end into the dispenser port on the rear of the reader.
13. Locate the injector-tip-cleaning stylus, packaged in a small cylinder. Attach the cylinder to the back of the dispenser for storage.
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20 | Chapter 2: Installation
9: Connect the Host Computer
The Synergy HTX is equipped with two types of communication ports: Serial (RS-232) and USB. Both ports are located on the rear panel of the reader.
· Both types of cables are included in the accessories box. Determine which cable is supported by the host computer.
· Connect one end to the appropriate port on the reader (see photo below) and the other end to the appropriate port on the host computer.
Figure 8: RS-232 serial and USB ports on the rear panel (injector model shown)
10: Install Gen5 Software
The Synergy HTX is controlled by BioTek’s Gen5 software running on a host computer. There is a certain sequence of events that must be followed to ensure that the software is properly installed and configured. Please follow the instructions provided in the Gen5 Getting Started Guide to install the software.
11: Turn on the Reader
Locate the power switch on the front panel and turn on the Synergy HTX. The reader will automatically initiate a System Test and eject the microplate carrier.
BioTek Instruments, Inc.

12: Establish Communication | 21
Figure 9: Carrier eject button (top) and power ON/OFF switch
12: Establish Communication
Important: If you are using the USB cable, refer to the instructions that shipped with the USB Driver Software to install the necessary drivers and identify the Com Port number.
1. Start Gen5 and log in if prompted. The default System Administrator password is admin.
2. From the Task Manager, select Setup > Go to System Menu. 3. Select System

Instrument Configuration, and click Add. 4. Set the Reader Type to Synergy HTX. 5. Set the Com Port to the computer’s COM port to which the reader is 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)).
6. 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.
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22 | Chapter 2: Installation
If the communication attempt is not successful, try the following: · Is the reader connected to the power supply and turned on? · Is the communication cable firmly attached to both the reader and the computer? · Did you select the correct Reader Type in Gen5? · Try a different COM port. · If using the USB cable, did you install the driver software? If you remain unable to get Gen5 and the reader to communicate with each other, contact BioTek’s Technical Assistance Center.
13: Set the Dispenser Calibration Values
Applies only to Synergy HTX models with injectors. Before you use the dispenser with the Synergy HTX, you must set the calibration values in Gen5.
1 Power on the instrument, and establish communication. 2 In Gen5, go to System > Instrument Configuration, select the Synergy HTX,
and click View/Modify. 3 Click Setup, and then select the Dispenser 1 tab. 4 On the keyboard, press CTRL+SHIFT+M to enter maintenance mode for the
Dispenser 1 window. 5 Enter the syringe calibration values from the label on the rear of the dispenser box. 6 Click Send Volumes, and then click Get Volumes to verify that the entered values
were sent to the instrument. 7 Select the Dispenser 2 tab, and repeat steps 4 through 6 for Dispenser 2.
BioTek Instruments, Inc.

Dispenser 1 calibration values

14: Run a System Test | 23
Dispenser 2 calibration values

Please contact BioTek’s Technical Assistance Center with any questions, tac@biotek.com.
14: Run a System Test
Running a System Test will confirm that the reader is set up and running properly, or will provide an error code if a problem has been detected.
1. Select System > Diagnostics > Run System Test. If prompted to select a reader, select the Synergy HTX and click OK.
2. When the test is complete, a dialog will appear requesting additional information. Enter the information (if required) and click OK.
3. The results report will appear, with text that reads “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. If the problem is something you cannot fix, or if the test continues to fail, contact BioTek’s Technical Assistance Center.
Synergy HTX Operator’s Manual

24 | Chapter 2: Installation

4. Models with injectors: Keep the software open and proceed to 15: Test Injector System.

All other models:

The installation and setup process is complete! Close the software and turn to page 26 to read about Operational/Performance Qualification.

15: Test the Injector System
Applies only to Synergy HTX models with injectors. 1. If necessary, press the button above the power switch to eject the microplate carrier. 2. Place the tip priming trough in the left-rear pocket of the carrier. 3. Place the priming plate on the carrier.

tip priming trough

Figure 10: Installing the tip priming trough and priming plate on the microplate carrier
4. 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.
BioTek Instruments, Inc.

15: Test the Injector System | 25 The dispense module’s setup should resemble the photo in Figure 11.
Make any final adjustments, if necessary.
Figure 11: The fully assembled dispense module
5. Select System > Instrument Control > Synergy HTX. 6. Click the Prime tab. 7. With Dispenser set to 1, set the Volume to 5000 µL and click Prime.
The syringe should move down and up repeatedly, drawing fluid from the bottle. The fluid should pump through the 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. 8. When the prime finishes, set Volume to 2000 µL and click Purge to clear the fluid lines. 9. Set Dispenser to 2 and repeat steps 7 and 8. 10. When finished, remove and empty the priming plate. 11. Close the software.
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26 | Chapter 2: Installation
Operational/Performance Qualification
Your Synergy HTX Multi-Detection Microplate Reader was fully tested at BioTek prior to shipment and should operate properly following the successful completion of the installation and setup procedures described throughout 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 HTX.
An Installation-Operational-Performance Qualification (IQ/OQ/PQ) package for the Synergy HTX is available for purchase (PN 1340508). Contact your local BioTek dealer for more information.
Repackaging and Shipping Instructions
Warning! If the reader and/or dispense module has been exposed to potentially hazardous material, decontaminate it to minimize the risk to all who come in contact with the reader during shipping, handling and servicing. Decontamination prior to shipping is required by the U.S. Department of Transportation regulations. See Appendix A for decontamination instructions. Caution! Remove the microplate and tip prime trough (if equipped) from the carrier before shipment. Spilled fluids can contaminate the optics and damage the instrument.
BioTek Instruments, Inc.

Repackaging and Shipping Instructions | 27

Important!
The instrument’s packaging design is subject to change. If the instructions in this section do not appear to apply to the packaging materials you are using, please contact BioTek’s Technical Assistance Center for 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 HTX 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 and can void the warranty.
The shipping materials are designed to be used no more than five times. If the original materials have been damaged, lost, or used more than five times, contact BioTek to order replacements (PN 7093001 for the reader, PN 7083001 for the dispense module). See page 6 for contact information.

Perform these steps to prepare the reader for shipment:
1. Contact BioTek’s Technical Assistance Center for a service authorization number before returning equipment for service. See page 6 for contact information.
2. Decontaminate the reader and, if attached, the dispense module, according to the instructions provided in Chapter 5.
3. If you will also be shipping the dispense module, perform these steps now:
a. With the reader on, start Gen5 and select System > Instrument Control > Synergy HTX.
b. Click the Prime tab. Ensure that “Dispenser” is set to 1.
c. Click the Maintenance button.
d. The Syringe 1 bracket will lower. Remove the thumbscrew from underneath the bracket. Carefully unscrew the top of the syringe from the syringe valve. Lift out the syringe and store it in its original box.
e. Set the Dispenser number to 2. Repeat steps c and d for Syringe 2.
f. Fully detach the dispense module from the reader. Replace the two nylon screws into the Syringe 1 and 2 tubing ports on the rear of the reader. (The screws should be stored in the plastic bag attached to the back of the module.) Set the module aside for the moment.

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28 | Chapter 2: Installation
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. 12). 6. Replace the microplate carrier shipping screw (see p. 11). 7. Tip the reader onto its back feet. Attach the shipping panel to the bottom of the
reader using the four screws and washers (see p. 10). 8. Wrap the plastic bag around the reader and shipping panel. 9. Locate the original outer shipping box. Place four foam blocks in the four
bottom corners of the box. Place the inner shipping box inside the outer box (see p. 8 and 9). 10. Grasp the handles on the shipping panel and carefully lower the reader into the inner shipping box. 11. Slide the foam vertical supports into place around the reader. Place the accessories box on top. 12. Close and seal the inner box with tape. 13. Place four foam corner blocks around the inner shipping box. Close and seal the outer box with tape. 14. Write the service authorization number in large, clear numbers on the outside of the box. Ship the box to BioTek.
Perform these steps to prepare the dispense module for shipment: 1. If you have not already done so: · Contact BioTek’s Technical Assistance Center for a service authorization number and shipping address before returning equipment for service. See page 6 for contact information. · Decontaminate the module according to the instructions in Chapter 5. · Remove the two syringes (see step 3 on the previous page) and store them in their original boxes. · Detach the dispense module outlet tubes and communication cable from the reader. Replace the two nylon screws into the Syringe 1 and 2 tubing ports on the rear of the reader.
Refer to the illustrations in 7: Unpack and Inspect the Dispense Module starting on page 15 when performing these steps.
2. Remove the two inlet tubes from the syringe valves and store them in their plastic canisters.
3. Remove the two outlet tubes from the syringe valves. Attach the clear plastic fitting covers to the fittings of the outlet tubes. Place the tubes in a plastic bag.
BioTek Instruments, Inc.

Repackaging and Shipping Instructions | 29 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 foam end cap into the box. 6. Place the syringes, the inlet tubes, and the outlet tubes inside the cutouts of the bottom foam end cap in the accessories box. Place the dispense module cover on top of the accessories. 7. Cover the accessories with the top foam end cap, place the dispense module cable inside the top of the end cap, and seal the box with tape. 8. Locate the original outer shipping box and foam end caps. Insert the bottom foam end cap. Lower the dispense module box into the end cap. 9. Insert the accessories box alongside the dispense module box. 10. Insert the top foam end cap. Close and seal the outer box with tape. 11. Write the service authorization number in large, clear numbers on the outside of the box. Ship the box to BioTek.
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30 | Chapter 2: Installation
BioTek Instruments, Inc.

Chapter 3
Getting Started
This chapter describes some of the Synergy HTX’s key components and provides an introduction to using Gen5 to control the instrument.
Key Components ………………………………………………………… 32 Power Switch, Carrier Eject Button, Microplate Carrier……….. 32 Lamp Assembly and Filter Wheel Access………………………… 33 Excitation and Emission Filter Wheels …………………………… 34 Installing the Time- Resolved Fluorescence Cartridge …………. 37 Configuring the System for Luminescence Measurements …… 38 The External Dispense Module ……………………………………. 39
Gen5 Software …………………………………………………………… 41 Viewing/Updating the Filter and Wavelengths Tables…………. 42 Creating Protocols and Experiments……………………………… 43 Controlling the Dispense Module …………………………………. 45
Recommendations for Optimum Performance ……………………… 48 Incubation and Partial Plates ……………………………………… 49

32 | Chapter 3: Getting Started
Key Components
Power Switch, Carrier Eject Button, Microplate Carrier
microplate carrier access door
filter wheel and lamp access door
carrier eject button power switch
Figure 12: Power switch, carrier eject button, microplate carrier
· The power switch contains an LED, which is illuminated green when the power is on.
· The microplate carrier eject button can be used to move the microplate carrier into or out of the measurement chamber and also to stop the instrument from “beeping” when it encounters an error.
· The microplate carrier supports microplates and adapter plates as described in Appendix A, Specifications. The plate is positioned so that well A1 is in the left rear corner of the carrier. A spring clip holds the plate 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 on 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. When the read is complete, the plate carrier slides to its full-out position.
BioTek Instruments, Inc.

Key Components | 33

For fluorescence and luminescence reading modes, the height of the top optical probe can be adjusted. Use the Read Height option to define how far the top probe shall be offset from the top surface of the plate during the read. In Gen5, this option is found in a Read step within a Procedure. Refer to the Gen5 Help for further instructions.

Lamp Assembly and Filter Wheel Access

Applies only to Synergy HTX models with fluorescence and luminescence capability.

Excitation filter wheel (TRF cartridge, when used)

Emission filter wheel

Fluorescence lamp assembly
Figure 13: Accessing the fluorescence lamp assembly and filter wheels
· The fluorescence lamp assembly and the excitation and emission filter wheels are accessible via a hinged door on the front of the instrument. To open the door, slip your finger into the notch on the right side and pull the door downward. A diagram showing the location of the lamp assembly and the orientation of the excitation and emission filter wheels is printed on the inside of the hinged door.
· For models with the Time-Resolved Fluorescence feature, remove the excitation filter wheel and replace it with the “TR” cartridge before running a time-resolved fluorescence assay. See page 37 for more information on the TR cartridge.

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34 | Chapter 3: Getting Started
The Synergy HTX has two lamps: one for standard fluorescence, 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 displays an error message. The lamp (PN 7080500) should be replaced at this time. Keeping a spare lamp on hand is recommended. Absorbance and Time- Resolved Fluorescence: This bulb should outlive the useful life of the reader. If there is a problem with the lamp, however, the intensity may drop and the run-time self-check will detect a low signal level and generate an error message. If this happens, the instrument will require service. Contact BioTek for assistance (this lamp is not user-replaceable).
Excitation and Emission Filter Wheels
Synergy HTX models with fluorescence capability are equipped with one excitation filter wheel and one emission filter wheel; readers with luminescence capability use an emission filter wheel only. (A monochromator is used for absorbance measurements.) A filter in the excitation wheel selects the narrow band of light to which the sample will be exposed. A filter in the emission wheel selects the band of light with the maximum fluorescence signal, to be measured by the photomultiplier (PMT). 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.
Each filter has its wavelength and bandpass values printed on its side, with an arrow to indicate the proper direction of light through the filter.
We recommend placing filters in the wheels in ascending wavelength order from position 1 to 4 (no holes in EX2 or EM3), particularly if the reader has generated a 4E18 (saturation) error.
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Supporting metal bracket
Thumbscrew

485/20

Key Components | 35
Direction of light
EXCITATION Filter wheel

Direction of light

528/20

EMISSION Filter wheel

Note the difference in filter orientation between the excitation and emission filter wheels

Figure 14: Profiles of the excitation and emission filter wheels, showing proper filter orientation

Important! The Synergy HTX is shipped with a set of excitation and emission filters installed, and the Synergy HTX’s onboard software is preconfigured with the filter values and their locations.

If you change the contents of a filter wheel, you must update Gen5’s filter table and then download the information to the reader. The Synergy HTX does not automatically detect which filters are installed.

See page 42 for information on updating Gen5’s filter table.

Removing the Filter Wheels
The filter wheels can be removed if different filter wheels need to be installed. It is important to note that:

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· The excitation and emission filter wheels are not interchangeable and are labeled as follows: EX = Excitation, EM = Emission. (TR = Time-Resolved Cartridge; see page 37.)
· 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 orientation.

To remove a filter wheel:

1. Important! Turn off the instrument.

2. Open the filter wheel access door using the depression on the right side of the door.

3. Observe the two thumbscrews within the compartment. The left thumbscrew holds the excitation filter wheel in place; the right secures the emission filter wheel.

4. Remove the thumbscrew and slide the filter wheel’s supporting metal bracket straight out of the compartment. Note: The emission filter wheel will “spring” out when removed. (This is because a shutter behind the wheel closes quickly to protect the PMT.)

Important! When removing or replacing a filter or C-clip filter retainer, do not use a sharp instrument! Use several layers of lens paper and your finger to remove and replace filters and clips. Using a sharp instrument, such as a flat screwdriver, will scratch the filter surface and make it unusable.

Do not touch the filters with your bare fingers!

To remove a filter or plug:
1. Turn the filter wheel to align the desired filter with the hole in the supporting bracket.
2. Place the bracket on a flat surface, with the filter wheel facing down.
3. Prepare a multi-layered “cushion” of lens paper. Using your 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.

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Key Components | 37
2. Properly orient the filter or plug (see page 34), 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 top of the hole containing the new filter. Cover your finger with several layers of lens paper, and then push down on all sides of the Cclip until it sits flush against the filter.
4. Clean both sides of the filter with lens paper. To reinstall a filter wheel:
1. Ensure that all filters and/or plugs are inserted properly (see above). 2. Slide the filter wheel back into its chamber. 3. Replace the thumbscrew. 4. Close the front door. 5. Turn on the instrument.
Installing the Time-Resolved Fluorescence Cartridge
For Synergy HTX models that support time-resolved fluorescence, the “TR” cartridge must be installed in place of the excitation filter wheel before a TRF assay can be run. The TR cartridge allows light from the xenon flash bulb to be input to the fluorescence optical system within the Synergy HTX. Excitation wavelengths are selected by adjusting the monochromator from 200 to 999 nm in 1-nm increments, with a fixed bandwidth of 10 nm.
The Synergy HTX automatically detects the presence of the TR cartridge. At the start of a time-resolved fluorescence assay, the operator will be prompted to install the TR cartridge if it is missing.
1. Important! Turn off the instrument. 2. Open the filter wheel access door using the depression on the right side of
the door. Observe the two thumbscrews within the compartment. The left thumbscrew holds the Excitation filter wheel in place. See the figure on page 33. 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 and turn on the instrument. See page 43 for more information on creating Gen5 protocols.
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Figure 15: The “TR” cartridge, for time-resolved fluorescence assays
Configuring the System for Luminescence Measurements
· For best results when taking luminescence measurements, the excitation filter wheel should have no empty locations, and it should have at least one “plug” (also referred to as a “dummy filter”) installed to prevent light from reaching the samples. Remove the excitation filter wheel (see page 35) and examine its contents; ensure that there are no empty locations and there is at least one plug installed.
· If your tests require that the light emitted from the samples remain unfiltered, the emission filter wheel should have an empty location in it. Remove the 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 filter table. Select “PLUG” to indicate the presence of a plug and “HOLE” to indicate an empty location. Click Send Values to download the information to the reader.
Updating Gen5’s filter table; for complete instructions, see page 42. BioTek Instruments, Inc.

· When defining a filter set in a Read step in a Gen5 procedure, selecting “Hole” indicates the empty location in the emission filter wheel. See page 43 for information on Read steps and procedures.

Key Components | 39

The External Dispense Module
Applies only to Synergy HTX models with injectors. The dispense module pumps fluid from the supply bottles to injector heads located inside the instrument. Fluid is injected into one well at a time.

Figure 16: Dispense module components
· Two 250-µL syringes draw fluid from the supply bottles. · 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.
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· Three-way valves switch the syringe flow from the inlet tubes to the outlet tubes.
· Outlet tubes transport fluid from the syringes into the instrument, through the tubing ports on the Synergy HTX’s rear panel. The outlet tubes are opaque PTFE tubes with threaded fittings on each end that are used to deliver fluid from the syringes to the instrument.
Inside the Synergy HTX, two Teflon tubes transport fluid from the tubing ports on the rear of the 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 17: Close-up view of the injectors inside the instrument
The tubing and injectors should be cleaned at least quarterly. See Chapter 4, Preventive Maintenance for more information.

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Gen5 Software | 41
Priming the System
Before an assay requiring fluid dispense is run, the system should be fully primed with the reagent or other fluid used by the assay. At the start of the assay (and optionally at the start of each dispense to a well), an additional injector tip prime can be performed. The tip prime compensates for any fluid loss at the injector tip due to evaporation since the last dispense. All priming activities are controlled via Gen5 (see page 46). 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 of the carrier, and is used for performing the Tip Prime before dispensing. The trough holds up to 1.5 mL of liquid and must be periodically emptied and cleaned by the user.
tip priming trough
Figure 18: Priming plate and tip priming trough
Gen5 Software
BioTek’s Gen5 software supports all Synergy HTX reader models. Use Gen5 to control the reader and the dispense module, perform data reduction and analysis on the measurement
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42 | Chapter 3: Getting Started
values, print /export results, and more. This section provides brief instructions for creating experiments and reading plates. It also explains how to use Gen5 to perform some functions that are specific to the dispense module.
Viewing/Updating the Filter and Wavelengths Tables
If configured with fluorescence or luminescence capability, the Synergy HTX ships with a set of excitation and emission filters installed, and the reader’s onboard software is preconfigured with the filter values and their locations. When Gen5 establishes communication with the reader, it “asks” for this information and then stores it in a filter table on the computer. To view this table in Gen5, select System > Instrument Configuration, highlight the Synergy HTX reader, and click View/Modify. Click Setup and then click the Fluorescence/Luminescence tab.
Regarding the Absorbance Wavelengths table: The Synergy HTX 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 within a protocol’s Reading Parameters dialog.
To change the settings and download them to the instrument: 1. Select Band Pass, Plug, or Hole for the excitation and emission filter wheels. 2. For each filter type, enter the wavelength value and its accompanying bandwidth. (The bandwidth is printed on the side of each filter.) 3. When finished, click Send Values to download the information to the reader. (Clicking Get Values uploads information from the reader.) 4. Click OK to save the settings and close this dialog. The settings become available for selection in the Read step dialog in a Procedure.
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Gen5 Software | 43
Creating Protocols and Experiments
In Gen5, a protocol 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. You’ll run the experiment to read plates and analyze the data.
Figure 19: Defining the procedure within a Gen5 protocol
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Figure 20: An experiment (containing measurement data), based on a predefined protocol
The instructions below briefly describe how to create a simple protocol in Gen5. For more information, or if the instructions below do not match what you see in Gen5, refer to the Gen5 Getting Started Guide or Help system.
1. To create a new protocol, from the Task Manager, select Protocols > Create New.
2. Select Protocol > Procedure. If prompted to select a reader, select the Synergy HTX and click OK.
3. Select a plate type. The assay plate must match the plate type selected in Gen5.
Otherwise, the results of the read may be invalid. 4. Add steps to the procedure for shaking or heating the plate, dispensing
fluid, reading the plate, and more. Click Validate 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.
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Gen5 Software | 45
· 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 define a Kinetic read, place an Endpoint Read step inside a Kinetic Start/End loop.
5. Open the Plate Layout dialog and assign blanks, samples, controls, and/or standards to the plate.
6. Open the Data Reduction dialog to add data reduction steps. Categories include Transformation, Well Analysis, Curve Analysis, and more.
7. Create a report or export template, via one of the Report/Export Builder options.
8. Save the file with an identifying name.
The instructions below briefly describe how to create an Experiment based on an existing protocol and then read a plate. See Gen5’s Help system for complete instructions.
1. To create a new experiment, from the Task Manager click, Experiments > Create using an existing protocol.
2. Select the desired protocol and click OK. 3. Select Plate > Read or click the Read Plate icon. 4. Click OK when the Load Plate dialog appears. The plate will be read. 5. When the read is complete, measurement values will appear in Gen5. To
view them, select the desired data set (e.g., “528/20,645/40”) from the Data drop-down list. 6. If you have not already done so, save the file with an identifying name.
Controlling the Dispense Module
Applies only to Synergy HTX models with injectors. Gen5 is used to perform several dispense module-specific functions, including initializing, priming, and purging. Gen5 also contains certain configuration items that must be set before using the dispense module. Read the following sections to become familiar with these functions and configuration items.
Initialization
If the dispense module was connected to the reader before the reader was turned on, or if a System Test was run via Gen5, the dispense module should initialize automatically. If for any reason the module does not initialize automatically, you can initialize it from Gen5:
1. In Gen5, select System > Instrument Control > Synergy HTX and click the Prime tab.
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2. Select the desired Dispenser number (1 or 2) and click Initialize. 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 bottle with a sufficient volume of the fluid to be used for the prime and the assay. Insert the appropriate inlet tube into the bottle.
2. Important! Place the priming plate on the carrier. 3. In Gen5, select System > Instrument Control > Synergy HTX and
click the Prime tab. 4. Select the Dispenser number (1 or 2) associated with the supply bottle. 5. Enter the Volume to be used for the prime, from 5 to 5000 µL.
The minimum recommended prime volume is 1100 µL. 6. Select a prime Rate, in µL/second. 7. Click Prime to start the process. 8. When the process is complete, carefully remove the priming plate from the
carrier and empty its contents. If the priming plate is empty, the prime volume was too low.
Purge Utility
Gen5 provides a special utility to purge fluid from the dispense tubing and syringe by pumping the fluid in reverse, back into the supply bottle. To purge the dispense module:
1. In Gen5, select System > Instrument Control > Synergy HTX and click the Prime tab.
2. Select the Dispenser number (1 or 2) associated with the supply bottle. 3. Enter the desired purge Volume in µL. 4. Select a prime Rate in µL/second. 5. Click Purge to start the process.
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Gen5 Software | 47
Syringe Maintenance Position
Gen5 provides access to special syringe setup functions 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. Select System > Instrument Control > Synergy HTX and click the Prime 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-fromhome position. The syringe can then be disconnected from the drive bracket and unscrewed from the valve.
4. See “Install Dispense Module Components” in Chapter 2, Installation for information on installing/removing the syringes.

Important! Do not change the syringe positions or calibrate the dispensers unless instructed to do so as part of installation, upgrade, or maintenance.

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Recommendations for Optimum Performance
· Microplates should be perfectly 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 covers or seals when not reading the plate. Filter solutions to remove particulates that could cause erroneous readings.
· Before preparing your microplates, make sure the instrument is on and successfully communicating with the controlling software. You may want to run a System Test if the instrument has not been turned off/on in a few days. Design your Gen5 protocol in advance as well, to ensure that the intended reading parameters are used and to avoid any last-minute corrections.
· Although the Synergy HTX supports standard flat, U-bottom, and V-bottom microplates, the reader achieves optimum performance with optically clear, flatbottomed wells. See Appendix A, Specifications for more information on the supported plates.
· Non-uniformity in the optical density of the well bottoms can cause loss of accuracy, 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 µL per well in a 96-well plate and 25 µL in a 384-well plate.
· Dispensing solution into 384-well plates often traps air bubbles in the wells, which may result in inaccurate readings. A dual-wavelength reading method usually eliminates these inaccuracies; however, for best results, remove the air bubbles by degassing the plate in a vacuum chamber before reading.
· The inclination of the meniscus can cause loss of accuracy in some solutions, especially with small volumes. Agitate the microplate before reading to help bring this problem 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 will stabilize over time.
BioTek Instruments, Inc.

Recommendations for Optimum Performance | 49 · To keep the dispense system in top condition, flush and purge the fluid lines with
deionized (DI) water every day or upon completion of an assay run, whichever is more frequent. Some reagents may crystallize or harden after use, clogging the fluid passageways. 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 4, Preventive Maintenance for more information. · For models with injectors: When dispensing volumes less than or equal to 20 µL/well, we recommend specifying a tip prime volume that is equal to the dispense volume. For dispense volumes greater than 20 µL/well, we recommend a tip prime volume of 20 µL.
Incubation and Partial Plates
When performing a partial plate read that includes an incubation step, the following recommendations can reduce the effects of evaporation of your samples:
· Use microplate lids. · Fill unused wells with fluid. · Cluster your sample wells rather than spacing them throughout the plate. · Place your sample wells in the center of the plate. This placement may lead to less
evaporation than if you place the samples in wells on the edge of the plate.
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BioTek Instruments, Inc.

Chapter 4
Preventive Maintenance
This chapter provides step-by-step instructions for maintaining the Synergy HTX and external dispense module (if used) in top condition, to ensure that they continue to perform to specification.
Recommended Maintenance Schedule ………………………………. 52 Overview ……………………………………………………………… 52 Daily Cleaning for the Dispense Module …………………………. 52 Recommended Maintenance Schedule…………………………… 53
Warnings and Precautions …………………………………………….. 54 Cleaning Exposed Surfaces ……………………………………………. 55 Inspect/Clean Excitation and Emission Filters ……………………… 56 Flush/Purge the Fluid Path …………………………………………….. 57 Running a Dispense Protocol (Optional) …………………………….. 58 Empty/Clean the Tip Priming Trough ………………………………… 59 Clean the Priming Plate ………………………………………………… 59 Clean the Internal Components ………………………………………. 60
Required Materials ………………………………………………….. 61 Removing the Reader’s Shroud …………………………………… 62 Removing the Internal Tubes and Injector Heads …………….. 64 Cleaning the Internal Tubes and Injector Heads ………………. 67 Cleaning the Optical Probes ……………………………………….. 68 Cleaning the Reader’s Internal Surface …………………………. 76 Reassembling the Components …………………………………… 77 Performance Check …………………………………………………. 79

52 | Chapter 4: Preventive Maintenance
Recommended Maintenance Schedule
Overview
A general Preventive Maintenance (PM) regimen for all Synergy HTX models includes periodically cleaning all exposed surfaces and inspecting/cleaning the Excitation and Emission filters. For models with the external dispense module, additional tasks include flushing/purging the fluid path and cleaning the tip prime trough, priming plate, supply bottles, internal dispense tubing, and injector heads.
Daily Cleaning for the Dispense Module
To keep the dispense module and injectors in top condition, flush and purge the fluid lines with deionized (DI) water every day or upon completion of an assay run, whichever is more frequent. Some reagents may crystallize or harden after use, clogging the fluid passageways. 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. Perform a visual inspection of the dispensing accuracy before conducting an assay that requires a dispense step 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 reagent in the dispensing lines using a suitable cleaning solution (review the reagent’s package insert for specific recommendations). A daily cleaning regimen is the best way to ensure accurate performance and a long life for your instrument and dispense module. BioTek also recommends flushing the module with DI water before conducting the decontamination procedure described in Chapter 5, As- Needed Maintenance.
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Recommended Maintenance Schedule | 53

Recommended Maintenance Schedule
The following charts recommend Preventive Maintenance tasks and the frequency with which each task should be performed.
It is important to note that the risk and performance factors associated with your assays may require that some or all of the procedures be performed more frequently than presented in the schedule.

Task All models: Clean exposed surfaces Inspect/clean excitation and emission filters (if equipped)
Decontamination
Models with injectors only: Flush/purge the fluid path (Optional) Run Dispense protocol Empty/clean tip prime trough Clean priming plate Clean internal components tubing and injector heads Clean optical probes Clean internal surfaces

Page
55 56
see Chapter
5
57 58 59 59 67
68 76

Daily

Quarterly As Needed

before shipment or storage

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Warnings and Precautions

Warning! Internal Voltage. Turn off and unplug the instrument for all maintenance and repair operations.
Warning! Wear protective gloves when handling contaminated instruments. Gloved hands should be considered contaminated at all times; keep gloved hands away from eyes, mouth, nose, and ears.
Warning! Mucous membranes are considered prime entry routes for infectious agents. Wear eye protection and a surgical mask when there is a possibility of aerosol contamination. Intact skin is generally considered an effective barrier against infectious organisms; however, small abrasions and cuts may not always be visible. Wear protective gloves when handling contaminated instruments.
Important! Do not immerse the instrument, 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.
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 may obstruct the carrier path and cause the reader to produce an error.
Caution! The buildup of deposits left by the evaporation of spilled fluids within the read chamber can impact measurements. Be sure to keep System Test records before and after maintenance so that changes can be noted.
Caution! Models with injectors. Before removing the reader’s cover to expose internal parts, purge the dispense module, turn off the instrument, and disconnect the fluid line, power cable, and PC cable.
Warning! The fluorescence lamp assembly is hot when the instrument is powered on. If the instrument is on, turn it off and allow the lamp to cool down before attempting to replace it.

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Cleaning Exposed Surfaces

Cleaning Exposed Surfaces | 55

Important! Turn off and unplug the instrument for all cleaning operations.
Important! Do not immerse the instrument, spray it with liquid, or use a “wet” cloth. Do not allow the cleaning solution to run into the interior of the instrument. If this happens, contact BioTek’s Service Department.

Exposed surfaces may be cleaned (not decontaminated) with a cloth moistened (not soaked) with water or water and a mild detergent. You will need:
· Deionized or distilled water
· Clean, lint-free cotton cloths
· Mild detergent (optional) To clean the exposed surfaces:
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 the instrument as necessary.
Models with injectors: If the Tip Priming Trough overflows, wipe the carrier and the surface beneath the carrier with a dry cotton cloth. If overflow is significant, you may have to remove the shroud of the instrument to better access the surface beneath the carrier.
See page 62 for instructions on removing the shroud. See page 76 for instructions for cleaning the surface beneath the
carrier.

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Inspect/Clean Excitation and Emission Filters
Applies only to Synergy HTX models with fluorescence and luminescence capability. Laboratory air is used to cool the lamp, and the filters can become dusty as a result. The filters should be inspected and cleaned at least every three months. You will need:
· Isopropyl, ethyl, or methyl alcohol · Lens-cleaning tissue Do not touch the filters with your bare fingers! To inspect and clean the excitation and emission filters: 1. Turn off and unplug the instrument. 2. Pull down the hinged door on the front of the instrument. Observe the two
thumbscrews within the compartment. The left thumbscrew holds the excitation (EX) filter wheel in place; the right secures 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 effect. This may indicate deterioration due to moisture exposure over a long period of time. · If you have any concerns about the quality of the filters, contact your
BioTek representative. 4. Clean the filters using lens-cleaning tissue moistened with a small amount of
isopropyl, ethyl, or methyl alcohol. Ensure that the filters remain in their current locations. 5. Replace the filter wheel brackets in their respective positions and replace the thumbscrews. Close the hinged door.
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Flush/Purge the Fluid Path
Applies only to Synergy HTX models with injectors. At the end of each day that the dispense module is in use, flush the fluid path using Gen5’s priming utility. Leave the fluid to soak overnight or over a weekend, and then purge the fluid before using the instrument again.
This flushing and purging routine is also recommended before disconnecting the outlet tubes from the rear of the reader, and before decontamination to remove any assay residue prior to applying isopropyl alcohol or sodium hypochlorite.
To flush the fluid path: 1. Fill two supply bottles with deionized or distilled water. Insert the supply (inlet) tubes into the bottles. 2. Place the priming plate on the carrier. 3. From Gen5’s main screen, select System > Instrument Control. Select the appropriate reader if prompted. 4. Click the Prime tab and select Dispenser 1. 5. Set the Volume to 5000 µL. 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 the fluid from the system (see below) and then prime with the dispense reagent before running an assay.
To purge the fluid from the system: 1. Place the inlet tubes in empty supply bottles or a beaker. 2. Select System > Instrument Control. Select the appropriate reader if prompted. 3. Click the Prime tab and select Dispenser 1. 4. Set the Volume to 2000 µL. 5. Click Purge to start the process. When the purge is complete, repeat the process for Dispenser 2.
After purging the system, you may wish to run a quick Dispense protocol to visually verify the dispense accuracy. See the next page for instructions for creating the protocol.
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Running a Dispense Protocol (Optional)

Applies only to Synergy HTX models with injectors.
After flushing/purging the system (page 57) and before running an assay that requires dispense, take a moment to visually inspect the dispensing accuracy.

Use a DI H2O­Tween solution to check for dispense accuracy following maintenance: e.g., add 1 mL Tween 20 to 1000 mL of deionized water.

Select a Plate Type in the Protocol that matches the plate you are using.
1. Create a new protocol and then select Protocol > Procedure. 2. Add a Dispense step with the following parameters:
· Select Dispenser 1 · Set Tip Priming to Before this dispense step and Volume to 10 µL. · Set 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 partial plate read on just one well (e.g., A1) · Set the Detection Method to Absorbance · Set the Read Type to Endpoint · Set the Read Speed to Normal · Select any wavelength 5. Click OK to close the dialog and add the Read step to the list. 6. Click OK to close the Procedure. 7. Save the protocol with an identifying name, such as “Dispense Observation.” 8. Create a new experiment to run the Dispense Observation protocol. 9. Initiate the plate read and follow the prompts.

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Empty/Clean the Tip Priming Trough | 59
10. When the procedure is complete, visually assess the fluid level in the wells for accuracy. If the well volume appears to be unevenly distributed, clean the internal dispense tubes and injector heads as described in Cleaning Internal Components on page 60.
Empty/Clean the Tip Priming Trough
Applies only to Synergy HTX models with injectors. The tip priming trough is a small, removable priming cup located in the left rear of the microplate carrier, used for performing the Tip Prime. The trough holds about 1.5 mL of liquid and must be periodically emptied and cleaned by the user. To empty/clean the tip prime trough:
1. Extend the microplate carrier and carefully remove the tip priming trough from its pocket in the left rear of the carrier.
2. Wash the trough in hot, soapy water. Use a small brush to clean in the corners. 3. Rinse the 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.
Clean the Priming Plate
Applies only to Synergy HTX models with injectors. Clean the priming plate regularly to prevent bacteria growth and residue buildup. Wash the plate in hot soapy water, using a small brush to clean in the corners if necessary. Rinse thoroughly and allow it to dry completely.
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Clean the Internal Components
Applies only to Synergy HTX models with injectors. The Synergy HTX’s internal components that require routine cleaning include:
· Optical probes · Surface beneath the microplate carrier · Internal dispense tubes and injector heads The internal components should be cleaned at least quarterly. In addition, if fluid has spilled inside the instrument and/or if an unusually high background signal has been flagged by the assay controls (typically blanks or negative controls), the optical probes and the surface 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 in Gen5 before and after performing these cleaning procedures. This will verify that all systems are functioning properly and allow you to compare results before and after maintenance.
Caution! The buildup of deposits left by the evaporation of spilled fluids within the read chamber can impact performance of both the fluorescence and absorbance functions. Be sure to perform a System Test before and after maintenance so that any changes in performance can be noted.
BioTek Instruments, Inc.

Required Materials

Clean the Internal Components | 61

Warning! Always wear protective gloves and safety glasses when performing cleaning/maintenance procedures.
For all tasks: · Protective gloves · Safety glasses
For removing the shroud and some of the internal components: · Screwdriver · 1/8″ hex key · 3/32″ hex key
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 optical probes:
· Clean cotton swabs · Isopropyl alcohol · Lens-cleaning tissue

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62 | Chapter 4: Preventive Maintenance
Removing the Reader’s Shroud
Caution! Before removing the shroud: Purge the dispense module (see page 57 for instructions), and then turn off and disconnect the reader from its power supply, the PC, and the dispense module.
The reader’s shroud (cover) must be removed to expose the internal components.

1. If you have not already done so, purge the dispense module of fluid. 2. Clear the work surface around the reader so you can easily access all sides of the instrument. 3. Disconnect power and all cables. Set the dispense module aside. 4. Remove four black mounting screws: one at the bottom rear corner on each side, and two at the top center of the rear panel.
   Remove two screws from the top center of the rear panel
   Remove two side screws When reinstalling the shroud, press down firmly on the top to maintain a good seal while tightening the top screws.
   BioTek Instruments, Inc.

Clean the Internal Components | 63 5. 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.
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64 | Chapter 4: Preventive Maintenance

Removing the Internal Tubes and Injector Heads
Take a moment to identify the components described in this section:

Internal tubes (2, black) for delivering fluid to the injector
heads

Tubing ports (2) for connecting the two internal dispense tubes to
the two external dispense tubes

Cable clamp for holding the tubes in place
Injector heads (2, white) for dispensing
fluid into microwells

Figure 20: Internal components for the injection system

Important! When reinstalling the 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

BioTek Instruments, Inc.

Clean the Internal Components | 65 Perform these steps to remove both sets of internal dispense tubes 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.
Cable clamp
3. Locate the injector heads. Turn each tube’s thumbscrew counterclockwise to disconnect the tube from the injector head.
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66 | Chapter 4: Preventive Maintenance 4. Turn the injector heads counterclockwise and gently pull them out of their sockets.
Be sure to seat the injector tips securely when reinstalling. See the photo on page 77.
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Clean the Internal Components | 67
Cleaning the Internal Tubes and Injector Heads
As discussed on page 52, some reagents can crystallize and clog the tubing and injector heads. Daily flushing and purging can help to prevent this, but more rigorous cleaning may be necessary if reagent has been allowed to dry in the tubing and/or injectors.
To clean the tubes: 1. Soak the internal tubes in hot soapy 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 into each injector head pipe to clear any blockages. (The stylus should be stored in a plastic cylinder affixed to the rear of the dispense module or reader.)
2. Stream water from a faucet through the pipe to be sure it is clean. If the water does not stream out, try soaking the heads in hot soapy water and then reinserting the stylus.
O-ring; do not remove
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68 | Chapter 4: Preventive Maintenance
Cleaning the Optical Probes
The optical probes should be cleaned at least quarterly. They should also be cleaned if reagent has spilled and/or if an unusually high background signal has been flagged by the assay controls (typically blanks or negative controls). Contaminated probes can lead to a loss of sensitivity (e.g., instead of being able to meet the 10 pg/mL concentration detection limit, the instrument may only be able to meet 20 pg/mL). Another indicator is the %CV in the Corners liquid test–it may increase due 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 62 now for instructions.
· We recommend cleaning the internal tubes and injector heads along with the optical probes. Instructions for removing and cleaning these components are provided on pages 64 through 67.
· Before starting this procedure, gather some supplies: Small container of isopropyl alcohol Small container of deionized or distilled water Lens- cleaning tissue Cotton swabs
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Clean the Internal Components | 69

Take a moment to identify the components discussed in this section:

Thumbscrew to access
ground wire

Heater/thermistor connectors (2)

Thumbscrews (2, black) for holding the incubator
housing in place

Incubator housing

Top optical probe
Top probe hanger, with two shoulder screws

Screw for lowering and raising the top
optical probe

Figure 21: Internal components to be removed/adjusted for cleaning the optic probes

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70 | Chapter 4: Preventive Maintenance Once the shroud has been removed and the internal tubes and injector heads have been removed and cleaned (see page 67), 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.
Depress tab to separate the connectors
2. Remove the thumbscrew located in the left rear of the instrument and set it aside. This exposes the ground wire.
BioTek Instruments, Inc.

Clean the Internal Components | 71 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.
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72 | Chapter 4: Preventive Maintenance 5. Turn the top probe screw counterclockwise to lower the probe hanger all the way to the bottom. (Rotate the screw, not the ring around it.)
Rotate counterclockwise
6. Gently lift the left side of the incubator housing and carefully slide it out.
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.
BioTek Instruments, Inc.

Clean the Internal Components | 73 7. Use a 1/8″ hex key 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 end of the probe in alcohol for one minute maximum. Wipe with lens-cleaning tissue and set aside.
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74 | Chapter 4: Preventive Maintenance
Clean with alcohol and lens tissue
9. Use a 3/32″ hex key to remove the two shoulder screws securing the top probe hanger. Remove the screws and set them aside.
Shoulder screws
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.
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Clean the Internal Components | 75

Probe hanger (bottom view)

Absorbance Lens

Important! When cleaning the absorbance lens with the swab, apply very little pressure to the lens! Applying too much 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.
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Cleaning the Reader’s Internal Surface
1. If you have not already done so, unplug the instrument and remove its shroud (see page 62 for instructions). Follow the instructions under Cleaning the Optical Probes to (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.
Microplate carrier, fully extended
BioTek Instruments, Inc.

Clean the Internal Components | 77 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. Wipe the instrument’s horizontal surface.
4. If detergent was used, wipe the surfaces with a cloth moistened with water. 5. Use a clean, dry, lint-free cloth to dry all wet surfaces.
Reassembling the Components
Perform these steps in the order listed to reassemble the components. Refer to the page numbers shown for further instructions and photos demonstrating the steps.
1. Slide the microplate carrier back into the instrument, page 76. 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.
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78 | Chapter 4: Preventive Maintenance
3. Attach the two internal dispense tubes to the injector heads, as shown below. Do not overtighten the thumbscrews!
Here is the top probe hanger ready for reinstallation, with injector heads and internal dispense tubes attached:
4. Replace the top probe hanger and shoulder screws (using the 3/32″ hex key), page 74.
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Clean the Internal Components | 79
5. Insert the top optic probe into its socket and replace its holding screw (using the 1/8″ hex key), page 73.
6. Replace the incubator housing and two thumbscrews, pages 72 and 71. Do not slide the two “forks” on the housing’s right side under the fixed foam housing.
7. Replace the groundwire and its thumbscrew, page 70. 8. Reconnect the heater and thermistor wires, page 70. 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, page 65. 10. Slide the two internal dispense tubes into the cable clamp and close the
clamp, page 65. 11. Review the steps you just performed to make sure the components have
been properly reassembled. 12. Slide the shroud onto the instrument, page 63. 13. Replace the four screws to securely attach the shroud to the base, page 62.
Performance Check
After reassembling the instrument, perform the following to verify that the instrument is functioning properly:
· Plug the instrument in and turn it on; allow its run-time system test to complete. Run a System Test through Gen5.
· Run any required OQ/PQ tests.
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BioTek Instruments, Inc.

Chapter 5
As-Needed Maintenance
This appendix contains procedures for decontaminating all models of the Synergy HTX. Purpose …………………………………………………………………… 82 Required Materials………………………………………………………. 83 Procedure for Models without Injectors ……………………………… 84 Routine Procedure for Models with Injectors ……………………….. 85
Clean Exposed Surfaces ……………………………………………. 85 Decontaminate the Fluid Lines ……………………………………. 86 Rinse the Fluid Lines ……………………………………………….. 87 Clean the Internal Tubing and Injector Heads …………………. 87 Clean the Tip Priming Trough and Priming Plate ………………. 88 Alternate Procedure for Models with Injectors ……………………… 89

82 | Chapter 5: As-Needed Maintenance

Purpose
Any laboratory instrument that has been used for research or clinical analysis is considered a biohazard and requires decontamination prior to handling. Decontamination minimizes the risk to all who come into contact with the instrument during shipping, handling, and servicing. Decontamination is required by the U.S. Department of Transportation regulations. Persons performing the decontamination process must be familiar with the basic setup and operation of the instrument.

BioTek Instruments, Inc. recommends the use of the following decontamination solutions and methods based on our knowledge of the instrument and recommendations 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 gloved hands away from eyes, mouth, and nose. Eating and drinking while decontaminating instruments is not advised.
Mucous membranes are considered prime entry routes for infectious agents. Wear eye protection and a surgical mask when there is a possibility of aerosol contamination. Intact skin is generally considered an effective barrier against infectious organisms; however, small abrasions and cuts may not always be visible. Wear protective gloves when performing the decontamination procedure.

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Required Materials | 83
Required Materials
For all Synergy HTX models: · Sodium hypochlorite (NaClO, or bleach) · 70% isopropyl alcohol (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: · Screwdriver · Small brush for cleaning the tip priming trough and priming plate · (Optional) Mild detergent
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Procedure for Models without Injectors

The sodium hypochlorite (bleach) solution is caustic; wear gloves and eye protection when handling the solution. Do not immerse the instrument, spray it with liquid, or use a “wet” cloth. Do not allow the cleaning solution to run into the interior of the instrument. If this happens, contact the BioTek Service Department.
Important! Turn off and unplug the instrument for all decontamination and cleaning operations.

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, 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 plate carrier door; slide out the plate carrier.
5. Wipe the plate carrier and all exposed surfaces of the instrument.
6. Wait 20 minutes. Moisten a cloth with deionized (DI) or distilled water and wipe all surfaces of the instrument that have been cleaned with the bleach solution or alcohol.
7. Use a clean, dry cloth to dry all wet surfaces.
8. Reassemble the instrument as necessary.
9. Discard the used gloves and cloths using a biohazard trash bag and an approved biohazard container.

BioTek Instruments, Inc.

Routine Procedure for Models with Injectors | 85
Routine Procedure for Models with Injectors
Perform this Routine Procedure when all systems are functioning normally on the Synergy HTX with Injectors. If you are unable to prime the Synergy HTX due to a system failure, perform the Alternate Procedure described on page 89.
If disinfecting with sodium hypochlorite (bleach), be sure to flush repeatedly with deionized water to ensure that no bleach is carried over. After disinfecting with sodium hypochlorite, perform the rinse procedure provided on page 87. If disinfecting with alcohol, do not immediately prime with deionized water, because the drying effect of the alcohol is an important aspect of its disinfectant properties.
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, prepare a 1:20 dilution. Household bleach is typically 5.0% NaClO; if this is the case, prepare a 1:10 dilution.
3. Manually open the plate carrier door; slide out the plate carrier. 4. Moisten a cloth with the bleach solution or alcohol. Do not soak the cloth. 5. Wipe the plate carrier and the exposed surfaces of the external dispense
module. 6. Wait 20 minutes. Moisten a cloth with deionized (DI) or distilled water and
wipe all surfaces that have been cleaned with the bleach solution or alcohol. 7. Use a clean, dry cloth to dry all wet surfaces.
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86 | Chapter 5: As-Needed Maintenance
8. Reassemble the instrument as necessary. 9. If the dispense module is installed, detach the outlet tubes from the rear panel
of the instrument. If it is not installed, attach just the dispense module’s communication cable to the instrument. Remove the supply bottles and their holders. 10. Perform the procedures described below through page 88 to decontaminate the fluid lines in the dispense module, the internal tubing and injector heads, and the tip priming trough and priming plate.
Decontaminate the Fluid Lines
1. Place a beaker with 20 mL of 0.5% sodium hypochlorite solution or 70% isopropyl alcohol near SYRINGE 1 on the dispense module.
2. Place the SYRINGE 1 inlet tube in the beaker. 3. If you have not already done so, detach the dispense 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, select System > Instrument Control, and click the Prime 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 presence). 7. Run two prime cycles, for a total of 10000 µL. 8. Pause for 20 to 30 minutes to allow the solution to disinfect the tubing. 9. Remove the inlet tube from the beaker of disinfectant solution. 10. From the Reader Control dialog, change the Volume to 1000 µL. 11. Run one prime cycle, to flush the disinfectant out of the fluid lines. 12. Empty the beaker containing the outlet tubes. Put the tubes back in. 13. Important! If sodium hypochlorite (bleach) was used, perform Rinse the Fluid Lines on the next page. Otherwise, (or after performing the Rinse procedure), repeat steps 1­13 for SYRINGE 2 / Dispenser 2.
BioTek Instruments, Inc.

Routine Procedure for Models with Injectors | 87
Rinse the Fluid Lines
Perform this procedure only if decontamination was performed using sodium hypochlorite. 1. Place a beaker containing at least 30 mL of deionized water on the dispense module. 2. Place the SYRINGE 1 or 2 inlet tube in the beaker. 3. If you have not already done so, place the outlet tubes in an empty beaker. 4. From the Reader Control dialog, select Dispenser 1 or 2, set the Volume to 5000 µL, and keep the default dispense Rate. 5. Run five prime cycles, for a total of 25000 µL. 6. Pause 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 Injector Heads
Turn to Chapter 4, Preventive Maintenance and perform the following procedures to access, remove, and clean the internal tubing and injector heads:
· Required Materials · Removing the Reader’s Shroud · Removing the Internal Tubes and Injector Heads · Cleaning the Internal Tubes and Injector Heads When finished, replace the internal components and the reader’s shroud.
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Clean the Tip Priming Trough and Priming Plate
1. Remove the tip priming trough from the left rear of the instrument’s microplate carrier (see below).
2. Wash the tip priming trough and priming plate in hot, soapy water. Use a small brush or cloth to clean 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 airdry.
5. Discard the used gloves and cloths using a biohazar

References

• UK.COM
• Chemical Analysis, Life Sciences, and Diagnostics | Agilent
• Chemical Analysis, Life Sciences, and Diagnostics | Agilent
• MEDISPEC
• Chemical Analysis, Life Sciences, and Diagnostics | Agilent
• Harta Instruments

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