Labcyte Echo 520, Echo 550, Echo 555 User Manual

For Echo models: 520

550 555

Echo Liquid Handler User Manual Labcyte Part Number 001-5331 Rev 6 March 2013

The information contained in this manual is subject to change without notice.

Any unauthorized changes or modifications to the Echo® liquid handler will void its warranty. Contact Labcyte Inc. prior to making any change or modification.

Labcyte and Echo are registered trademarks of Labcyte Inc.in the United States. Microsoft, Windows, Windows XP, Windows 7, ActiveX, Excel, and Microsoft SQL Server are either registered trademarks or trademarks of Microsoft Corporation in the United States and other countries. Intel and Intel Core 2 Duo, are trademarks or registered trademarks of Intel Corporation or its subsidiaries in the United States and other countries. Alconox is a registered trademark of Alconox, Inc.

All other registered or unregistered trademarks used herein are the exclusive property of their respective holders.

For research purposes only; not for use in diagnostics.

The Echo liquid handler is covered by one or more of the following patents:

6,416,164; 6,548,308; 6,603,118; 6,612,686; 6,642,061; 6,666,541; 6,707,038; 6,710,335; 6,746,104; 6,802,593; 6,808,934; 6,809,315; 6,849,423; 6,855,925; 6,869,551; 6,893,836; 6,893,115; 6,916,083; 6,932,097; 6,938,987; 6,938,995; 6,991,917; 7,070,260; 7,090,333; 7,354,141; 7,405,072; 7,717,544; 7,899,645; 7,900,505; 7,901,039; 8,107,319.

European Patent EP: 1337325; 1324823; 1337325; 1352112; 1366356; 1534526; 1585636.

Japan: 4189964; 4309131; 4434581; 4559218; 4624644; 4955901; 4990476

Additional patents pending in the United States and other countries.

ii Echo Liquid Handler User Manual

Contents

Preface

Intended audience . . . . . . . . . . . . . . . . . . . . . . . . . . vii

Safety warnings and precautions . . . . . . . . . . . . . . . . . . viii

Safety notation marks . . . . . . . . . . . . . . . . . . . . . viii

Electrical safety warnings . . . . . . . . . . . . . . . . . . . . . ix

Additional safety warnings . . . . . . . . . . . . . . . . . . . . x

Safety cautions . . . . . . . . . . . . . . . . . . . . . . . . . . x

Emergency Off switch (EMO). . . . . . . . . . . . . . . . . . . xii

Laser safety warnings . . . . . . . . . . . . . . . . . . . . . . . xii

Product labels . . . . . . . . . . . . . . . . . . . . . . . . . . . xiii

Intellectual property label . . . . . . . . . . . . . . . . . . . . xiv

ETL label . . . . . . . . . . . . . . . . . . . . . . . . . . . . xiv

Laser safety labels. . . . . . . . . . . . . . . . . . . . . . . . xiv

Electrical hazard . . . . . . . . . . . . . . . . . . . . . . . . xiv

General warning label . . . . . . . . . . . . . . . . . . . . . xiv

Water system maintenance label . . . . . . . . . . . . . . . . . xv

WEEE label . . . . . . . . . . . . . . . . . . . . . . . . . . . . xv

Emergency Off label . . . . . . . . . . . . . . . . . . . . . . . xv

Laser safety labels. . . . . . . . . . . . . . . . . . . . . . . . xvi

Product label . . . . . . . . . . . . . . . . . . . . . . . . . . xvi

Pinch point label . . . . . . . . . . . . . . . . . . . . . . . . xvi

High-voltage label . . . . . . . . . . . . . . . . . . . . . . . xvii

Using this manual . . . . . . . . . . . . . . . . . . . . . . . . . xviii

Who should read this manual? . . . . . . . . . . . . . . . . . xviii

What is in this manual?. . . . . . . . . . . . . . . . . . . . . xviii

Conventions used in this manual . . . . . . . . . . . . . . . . xix

Chapter 1 Introduction

1.1 Acoustic Droplet Ejection . . . . . . . . . . . . . . . . . . . . 1-2

1.1.1 How does the Echo system work? . . . . . . . . . . . . 1-2

1.1.2 Will the fluid splash or fall out? . . . . . . . . . . . . . 1-4

1.2 What are the Echo liquid handler features? . . . . . . . . . . . 1-5

1.2.1 “Touchless” fluid transfers . . . . . . . . . . . . . . . . 1-5

1.2.2 Reliability . . . . . . . . . . . . . . . . . . . . . . . . 1-6

1.2.3 Ease of use. . . . . . . . . . . . . . . . . . . . . . . . 1-7

1.2.4 System integration friendly. . . . . . . . . . . . . . . . 1-8

1.2.5 Low maintenance requirements . . . . . . . . . . . . . 1-9

1.2.6 Cost savings . . . . . . . . . . . . . . . . . . . . . . . 1-9

1.2.7 High accuracy, precision, and speed . . . . . . . . . . . 1-10

1.2.8 Value-added quality control . . . . . . . . . . . . . . . 1-11

1.3 What types of liquid transfer are supported? . . . . . . . . . . 1-12

1.3.1 384-plate to 384-plate liquid transfer . . . . . . . . . . 1-13

1.3.2 384-plate to 96-plate liquid transfer . . . . . . . . . . . 1-14

1.3.3 384-plate to 1536-plate liquid transfer. . . . . . . . . . 1-15

1.3.4 1536-plate to 384-plate liquid transfer. . . . . . . . . . 1-16

Contents iii

1.3.5 1536-plate to 1536-plate liquid transfer . . . . . . . . . 1-17

1.4 What system configurations are supported? . . . . . . . . . . 1-18

1.4.1 Manual operation . . . . . . . . . . . . . . . . . . . . 1-18

1.4.2 Echo applications . . . . . . . . . . . . . . . . . . . . 1-19

1.4.3 Modular workstation operation . . . . . . . . . . . . . 1-20

1.4.4 Small cell operation . . . . . . . . . . . . . . . . . . . 1-21

1.4.5 Fully automated operation . . . . . . . . . . . . . . . . 1-22

1.4.6 Multi-client considerations . . . . . . . . . . . . . . . . 1-23

Chapter 2 Installation and Relocation

2.1 Pre-installation requirements . . . . . . . . . . . . . . . . . . 2-2

2.1.1 Site requirements . . . . . . . . . . . . . . . . . . . . 2-2

2.1.2 Additional components . . . . . . . . . . . . . . . . . 2-4

2.2 Installation overview . . . . . . . . . . . . . . . . . . . . . . 2-5

2.3 Software installation . . . . . . . . . . . . . . . . . . . . . . 2-6

2.3.1 Install the software. . . . . . . . . . . . . . . . . . . . 2-7

2.3.2 Uninstall the Echo software . . . . . . . . . . . . . . . 2-9

2.3.3 Upgrade the Echo software . . . . . . . . . . . . . . . 2-9

2.4 Instrument Relocation . . . . . . . . . . . . . . . . . . . . . 2-10

2.4.1 Prepare the Echo liquid handler for a move . . . . . . . 2-10

2.4.2 Reinstall the Echo liquid handler after a move . . . . . . 2-11

Chapter 3 System Description

3.1 System overview . . . . . . . . . . . . . . . . . . . . . . . . 3-2

3.1.1 Front panel . . . . . . . . . . . . . . . . . . . . . . . 3-2

3.1.2 Back panel . . . . . . . . . . . . . . . . . . . . . . . . 3-4

3.2 Component description. . . . . . . . . . . . . . . . . . . . . 3-5

3.2.1 Process door . . . . . . . . . . . . . . . . . . . . . . . 3-5

3.2.2 Source plate gripper stage . . . . . . . . . . . . . . . . 3-6

3.2.3 Source plate insert . . . . . . . . . . . . . . . . . . . . 3-6

3.2.4 Destination plate gripper stage. . . . . . . . . . . . . . 3-8

3.2.5 Status indicator lights . . . . . . . . . . . . . . . . . . 3-9

3.2.6 LCD screen. . . . . . . . . . . . . . . . . . . . . . . . 3-11

3.2.7 Emergency Off switch . . . . . . . . . . . . . . . . . . 3-11

3.2.8 Anti-static bars. . . . . . . . . . . . . . . . . . . . . . 3-12

3.2.9 AC power and fuse compartment . . . . . . . . . . . . 3-14

3.2.10Coupling fluid bottle . . . . . . . . . . . . . . . . . . . 3-15

3.2.11Fluid chiller. . . . . . . . . . . . . . . . . . . . . . . . 3-16

3.2.12Surge tank . . . . . . . . . . . . . . . . . . . . . . . . 3-16

3.2.13Fluidics panel . . . . . . . . . . . . . . . . . . . . . . 3-17

3.3 Specifications . . . . . . . . . . . . . . . . . . . . . . . . . . 3-18

3.3.1 Physical . . . . . . . . . . . . . . . . . . . . . . . . . 3-18

3.3.2 Mechanical . . . . . . . . . . . . . . . . . . . . . . . 3-18

3.3.3 Environmental . . . . . . . . . . . . . . . . . . . . . . 3-19

iv Echo Liquid Handler User Manual

3.3.4 Electrical . . . . . . . . . . . . . . . . . . . . . . . . . 3-19

3.3.5 Client PC. . . . . . . . . . . . . . . . . . . . . . . . . 3-19

3.3.6 Communications. . . . . . . . . . . . . . . . . . . . . 3-19

3.3.7 Fluid transfer. . . . . . . . . . . . . . . . . . . . . . . 3-20

3.3.8 Supported labware. . . . . . . . . . . . . . . . . . . . 3-22

3.3.9 Coupling fluid . . . . . . . . . . . . . . . . . . . . . . 3-23

Chapter 4 Manual Operation

4.1 Startup/Shutdown . . . . . . . . . . . . . . . . . . . . . . . 4-2

4.1.1 Turn on Echo power . . . . . . . . . . . . . . . . . . . 4-2

4.1.2 Launch Echo software . . . . . . . . . . . . . . . . . . 4-4

4.1.3 Turn off Echo power . . . . . . . . . . . . . . . . . . . 4-5

4.2 Main screen . . . . . . . . . . . . . . . . . . . . . . . . . . 4-8

4.3 Create liquid transfer protocol . . . . . . . . . . . . . . . . . 4-10

4.3.1 Add new protocol . . . . . . . . . . . . . . . . . . . . 4-11

4.4 Run the liquid transfer protocol . . . . . . . . . . . . . . . . . 4-13

4.4.1 Prepare the microplates . . . . . . . . . . . . . . . . . 4-13

4.4.2 Start liquid transfer run . . . . . . . . . . . . . . . . . 4-14

Chapter 5 Echo Liquid Handler Software

5.1 Protocols window. . . . . . . . . . . . . . . . . . . . . . . . 5-2

5.1.1 Create a protocol . . . . . . . . . . . . . . . . . . . . 5-4

5.1.2 Edit an existing protocol definition . . . . . . . . . . . . 5-16

5.1.3 Remove an existing protocol definition. . . . . . . . . . 5-17

5.1.4 Run a protocol . . . . . . . . . . . . . . . . . . . . . . 5-17

5.2 Labware window . . . . . . . . . . . . . . . . . . . . . . . . 5-20

5.2.1 Plate name convention. . . . . . . . . . . . . . . . . . 5-21

5.2.2 Plate specification editor . . . . . . . . . . . . . . . . . 5-23

5.3 Diagnostics window . . . . . . . . . . . . . . . . . . . . . . 5-28

5.3.1 Device control . . . . . . . . . . . . . . . . . . . . . . 5-29

5.3.2 Status indicators . . . . . . . . . . . . . . . . . . . . . 5-33

5.3.3 Diagnostic procedures . . . . . . . . . . . . . . . . . . 5-38

5.4 Calibration window. . . . . . . . . . . . . . . . . . . . . . . 5-44

5.4.1 Motion calibration . . . . . . . . . . . . . . . . . . . . 5-45

5.4.2 Power calibration . . . . . . . . . . . . . . . . . . . . 5-47

5.4.3 Transducer calibration . . . . . . . . . . . . . . . . . . 5-50

5.5 Advanced window . . . . . . . . . . . . . . . . . . . . . . . 5-53

5.5.1 Advanced liquid transfer controls . . . . . . . . . . . . 5-53

5.5.2 Advanced solvent concentration (DMSO only) . . . . . . 5-55

5.5.3 Advanced Echo control. . . . . . . . . . . . . . . . . . 5-57

5.6 Status window . . . . . . . . . . . . . . . . . . . . . . . . . 5-58

Chapter 6 Maintenance and Service

6.1 Maintenance schedule . . . . . . . . . . . . . . . . . . . . . 6-2

Contents v

6.1.1 Daily maintenance . . . . . . . . . . . . . . . . . . . . 6-2

6.1.2 Echo maintenance alerts . . . . . . . . . . . . . . . . . 6-3

6.1.3 Scheduled maintenance . . . . . . . . . . . . . . . . . 6-5

6.2 Maintenance procedures . . . . . . . . . . . . . . . . . . . . 6-7

6.2.1 Maintenance tools and materials. . . . . . . . . . . . . 6-7

6.2.2 Refill the coupling fluid bottle . . . . . . . . . . . . . . 6-8

6.2.3 Clean and refill the coupling fluid bottle . . . . . . . . . 6-10

6.2.4 Empty the coupling fluid bottle . . . . . . . . . . . . . 6-11

6.2.5 Replace the water filter . . . . . . . . . . . . . . . . . 6-15

6.2.6 Clean the anti-static bars. . . . . . . . . . . . . . . . . 6-16

6.2.7 Replace the AC power fuse . . . . . . . . . . . . . . . 6-18

6.3 Extended Non-Use and Storage. . . . . . . . . . . . . . . . . 6-19

6.3.1 Maintenance During Extended Non-Use . . . . . . . . . 6-19

6.3.2 Preparation for storage. . . . . . . . . . . . . . . . . . 6-20

Chapter 7 Contact Information and Troubleshooting

7.1 Contact information . . . . . . . . . . . . . . . . . . . . . . 7-1

7.2 Setup problems. . . . . . . . . . . . . . . . . . . . . . . . . 7-2

7.3 Maintenance alerts . . . . . . . . . . . . . . . . . . . . . . . 7-3

Appendix A. Acoustic Droplet Ejection Technology

A.1 ADE history . . . . . . . . . . . . . . . . . . . . . . . . . . . A-1

A.2 ADE and the Echo liquid handler . . . . . . . . . . . . . . . . A-2

A.3 Source microplate survey . . . . . . . . . . . . . . . . . . . . A-3

A.4 Fluid transfer . . . . . . . . . . . . . . . . . . . . . . . . . . A-4

A.4.1 Positioning the ADE elements . . . . . . . . . . . . . . A-4

A.4.2 Creating the acoustic pulse . . . . . . . . . . . . . . . A-4

A.4.3 Transferring the droplet . . . . . . . . . . . . . . . . . A-5

A.4.4 Examples of acoustic droplet ejection . . . . . . . . . . A-6

Appendix B. Barcode Locations

B.1 Short flange height microplates. . . . . . . . . . . . . . . . . B-2

B.2 Medium flange height microplates . . . . . . . . . . . . . . . B-2

Appendix C. Chiller Information

C.1 Shipping contents . . . . . . . . . . . . . . . . . . . . . . . C-2

C.2 Safety Warnings and Precautions . . . . . . . . . . . . . . . . C-2

C.3 Chiller Setup and Operation . . . . . . . . . . . . . . . . . . C-3

C.3.1 Setup . . . . . . . . . . . . . . . . . . . . . . . . . . C-3

C.3.2 Operation . . . . . . . . . . . . . . . . . . . . . . . . C-4

C.4 Troubleshooting . . . . . . . . . . . . . . . . . . . . . . . . C-4

vi Echo Liquid Handler User Manual

The preface to the Labcyte® Echo® liquid handler user manual contains important information regarding the safe use of an Echo liquid handler and how to use this manual. Read and understand the safety information thoroughly before you begin operating the Echo liquid handler.

 Intended audience

 Safety warnings and precautions

 Barcode scanner locations

 Using this manual

The Labcyte Echo 520, 550 and 555 systems are referred to as the Echo liquid handler throughout this manual. The information and illustrations apply to all models unless specifically stated otherwise.

Intended audience

The Echo liquid handler is designed to be used by individuals who are familiar with good laboratory practices (GLP)1 or similar laboratory safety program.

CHAPTER 0PREFACE

Echo liquid handler operators must read this manual before using the Echo liquid handler instrument.

Throughout this manual the word “you” or “user” refers to the Echo liquid handler operator.

1 Good laboratory practice (GLP) for nonclinical laboratory studies, FDA regulation 21 CFR

Part 58.

Preface vii

Safety warnings and precautions

The Echo liquid handler has been designed for safe operation.

The safety warnings and precautions in this section and throughout the manual must be observed during installation, relocation, maintenance, repair, and normal operation of an Echo liquid handler.

Failure to comply with these warnings and precautions, or with specific cautions and warnings found elsewhere in this manual, violates the safety standards of design, manufacture, and intended use of an Echo liquid handler. This can result in hazardous exposure to laser light, high voltage, or moving parts. Exposure to these hazards can cause severe injury.

Safety notation marks

This manual uses the following symbols in the left margin to draw your attention to the specified type of information.

Symbol Meaning

Warning

Warnings alert all users to the following:

 Potentially hazardous conditions

 Actions that may result in personal injury or death

Caution

Cautions alert the user to actions that may result in the following:

 Damage to the equipment

 Lost or corrupted data

 Unrecoverable interruption of the operation being

performed

Note

Notes emphasize or expand upon the surrounding information.

viii Echo Liquid Handler User Manual

Electrical safety warnings

Always observe the following electrical safety warnings:

Warning: Plug the Echo liquid handler into a grounded circuit (Class 1) capable of delivering at least:

 15 A for a 100– 120 VAC~ power source.  10 A for a 200– 240 VAC~ power source.

If you are unable to insert an AC plug into your AC receptacle, contact an electrician to correct the situation.

Warning: The Echo liquid handler operates with voltages and currents that can be lethal. Pushing objects of any kind into the Echo liquid handler, through slots or holes in its covers, may cause serious electrical shock or may short out electrical circuits or parts. Do not spill any liquid inside or on the Echo liquid handler.

Warning: The Echo liquid handler contains user-replaceable AC power fuses. The fuse holder contains two fuses. Use only the specified replacement fuses and the fuse replacement procedure found at “Replace

the AC power fuse” on page 6-18. If a fuse requires repeated replacement,

the Echo liquid handler could have an electrical problem. Do not use a malfunctioning Echo liquid handler. Contact Labcyte Service and Support (see “Contact information” on page 7-1).

Warning: Do not use AC power cords if the following conditions exist:

 The power cords are frayed or damaged.  Other attached cords, cables, or receptacles are frayed or damaged.

Use of damaged power cords can cause an electrical shock hazard and result in severe injury.

Warning: Do not connect (plug in) or disconnect (unplug) AC power cords if the following conditions exist:

 The Echo liquid handler, or attached equipment, has been exposed to

excessive moisture, or to liquids that have been spilled on it.

 The Echo liquid handler, or any of its subassemblies or components,

has been dropped or damaged.

 You suspect service or repair is required.

 The Echo liquid handler is being cleaned, handled during a

maintenance procedure, or repaired—except as directed in the applicable written procedure.

These conditions can cause an electrical shock hazard and result in severe injury.

Preface ix

Additional safety warnings

Warning: Do not try to gain access to the interior of the Echo liquid handler. Do not remove the Echo liquid handler exterior covers. Exposure to laser light, high voltage, or moving parts inside the Echo liquid handler can cause severe injury.

Warning: Keep your hands, fingers, and clothing clear of the process door and microplate assemblies, except when loading or unloading a plate. Be aware of the possibility that either the source or the destination plate gripper stages may move.

Warning: Use good laboratory practices and follow the manufacturer’s precautions when working with chemicals. Labcyte is not responsible or liable for any damages as a result of, or as a consequence of, the use of hazardous chemicals.

Warning: The Echo liquid handler weighs approximately 128 kg (283 lbs). Moving or lifting the Echo liquid handler incorrectly can cause severe injury. Never attempt to move or lift the Echo liquid handler without using proper equipment and proper safety techniques. Contact the safety coordinator at your company for information.

Safety cautions

To protect the Echo liquid handler from damage, follow these precautions:

Caution: Always use the normal shutoff routine or the EMO (Emergency Motion Off) button to turn off the Echo instrument. Turning off the instrument by other methods (for example, unplugging the power cord), can potentially result in damage to the electronics or corrupt the software.

Caution: Do not attempt to service or repair the internal Echo liquid handler mechanisms yourself. The electrical, laser, and mechanical systems must be maintained by Labcyte field engineers. If you have any questions regarding what may be serviced by an Echo liquid handler user, please call Labcyte (see “

Caution: Do not make modifications to the Echo liquid handler.

Caution: Use only replacement parts that are approved by Labcyte.

Caution: Use labware that is SBS-compliant

Contact information” on page 7-1).

or approved by Labcyte.

2 Society for Biomolecular Sciences. See “Supported labware” on page 3-22.

x Echo Liquid Handler User Manual

Caution: Do not obstruct the air vents. Keep all air vents free of dirt or

dust.

Caution: Handle the Echo liquid handler coupling fluid and waste water carefully. Avoid all spills.

Caution: Keep corrosive agents, or otherwise damaging material, away from the Echo liquid handler and its attached devices.

Caution: Do not expose the Echo liquid handler to excessive moisture (>80% relative humidity). Moisture, or condensation, can damage electrical components.

Caution: Do not expose the Echo liquid handler to temperatures outside of the storage range: 5°C–45°C (41°F–113°F). Temperatures outside of this range can damage the instrument.

Caution: Do not run the Echo liquid handler without the chiller. Ensure that the chiller is correctly connected to the Echo instrument and is running at the recommended temperature. Incorrect coupling fluid temperature can reduce the precision and accuracy of the fluid that is transferred. Extreme fluid temperatures can potentially damage the acoustic transducer.

Caution: Do not allow the acoustic transducer to dry out. If the Echo liquid handler is going to be shut down for longer than a week, start up and run the pump once a week to keep the acoustic transducer wet. See

“Extended Non-Use and Storage” on page 6-19.

Caution: Do not run the pump if the chiller tubing is not connected. Running the pump without the chiller tubing may damage other system components.

Caution: Do not run the pump without the vacuum source connected and active. Running the pump without vacuum can lead to flooding the system or a system error.

Preface xi

Emergency Off switch (EMO)

Laser apertures

The Echo liquid handler includes two Emergency Off switches, also known as EMO (Emergency Motion Off) switches. They are located on the front and rear of the unit and are prominently labeled “EMERGENCY OFF.” See

“Emergency Off label,” on the next page for switch

location.

See “Emergency Off switch” on page 3-11 for a detailed explanation of

what the Emergency Off switch turns off, when to use it, how to use it, and how to resume normal Echo liquid handler operation after using it.

Laser safety warnings

The Echo liquid handler is a Class I laser instrument that can house up to three optional laser-based barcode scanners.

When operated as specified in this manual, the Echo liquid handler does not expose the operator to laser light. Nevertheless, during a barcode scan, the Class II diode lasers (with maximum power up to 1.0 mW at 650 nm) can be visible from the interior of the unit. The barcode scanners are not interlocked; therefore, if the Echo liquid handler is operated with the covers off, the operator may be exposed to a Class II laser light hazard. Do not operate the Echo liquid handler if its covers have been removed.

Figure 1 Barcode scanner locations

xii Echo Liquid Handler User Manual

Product labels



 















You will find the following labels on the Echo liquid handler.

1. Intellectual property label

2. ETL label

3. Laser safety labels

4. Electrical hazard

5. General warning label

6. Water system maintenance label

7. WEEE label

8. Emergency Off label

9. Laser safety labels

10. Product label

11. Pinch point labels

12. High-voltage label

Figure 2 Echo liquid handler label locations (applies to all models).

Preface xiii

Intellectual property label

The intellectual property label cites United States patents and other applicable legal protection.

ETL label

The ETL label indicates conformance to ETL SEMKO product safety standards for the US and Canada.

Laser safety labels

One of two laser safety labels (this one is visible on the instrument back cover).This warning indicates

that the Echo liquid handler optionally uses laserbased barcode scanners. The Echo liquid handler is considered a Class 1 laser light hazard under normal operating conditions (with instrument covers on).

Electrical hazard

General warning label

xiv Echo Liquid Handler User Manual

The electrical hazard label warns of possible injury from electrical cables, connections, and circuits inside the Echo liquid handler.

The general warning label reminds users to take normal safety precautions when operating and working around an Echo liquid handler.

Water system maintenance label

WEEE label

Waste Electrical and Electronic Equipment (WEEE) directive

The WEEE label signifies that this instrument should not be disposed of in the trash. Return it to the manufacturer for recycle or ecological disposal.

The water system maintenance label is provided by Labcyte to remind the user to follow recommended maintenance procedures. It also emphasizes the use of distilled water.

Emergency Off label

The Emergency Off label identifies the EMO (Emergency M

otion Off) switch, which is used in an emergency to shut down all mechanical activity in the Echo liquid handler. There are two Emergency Off switches, one on the front panel and one on the rear panel of the Echo liquid handler.

Preface xv

Laser safety labels

One of two laser safety labels (this label is visible above the opening when the front cover is removed), This warning indicates that the Echo liquid

handler optionally uses laserbased barcode scanners. The Echo liquid handler is considered a Class 1 laser light hazard under normal operating conditions (with instrument covers on).

Product label

The Echo liquid handler product label (located inside, beside the pump), contains Labcyte corporate identification, product identification, product serial number, regulatory agency marks, origin of manufacture, date of manufacture, input power specifications, and CE mark3.

Pinch point label

3 The CE mark indicates compliance with the EU (European Union) Directives.

xvi Echo Liquid Handler User Manual

The pinch point label warns of possible injury if you do not keep your hands or fingers clear of source plate or destination microplate assemblies when they are in motion.

High-voltage label

The high-voltage label warns of possible high-voltage injury from electrical cables, connections, and circuits inside the Echo liquid handler.

Preface xvii

Using this manual

Who should read this manual?

The Echo liquid handler user manual supports several types of users involved in liquid handling and liquid transfer.

 The Preface contains safety information that should be read by

everyone.

 Chapters 1, 3, 4, 5, and Appendix A and B should be read by

everyone.

 Chapter 2 is of special interest to those involved in installing or

relocating an Echo liquid handler.

 Chapter 6 should be read by everyone who is responsible for user-

level maintenance and interfacing with Labcyte service and support personnel.

What is in this manual?

The Echo liquid handler user manual contains the following:

 Chapter 1, “Introduction,” discusses the Echo liquid handler, its

several operating modes and applications, and how ADE (acoustic droplet ejection) is performed.

 Chapter 2, “Installation and Relocation,” covers pre-installation

requirements, software installation, and moving instructions.

 Chapter 3, “System Description,” describes the system components,

controls, external connections, status indicators, and the LCD display.

 Chapter 4, “Manual Operation,” takes the reader through the

performance of a sample fluid transfer protocol using the Echo liquid handler software.

 Chapter 5, “Echo Liquid Handler Software,” provides a thorough

description of software functions to: set up protocol and destination plate definitions, run fluid transfer protocols, perform diagnostic and calibration procedures, and set up advanced fluid transfer options.

 Chapter 6, “Maintenance and Service,” covers maintenance

schedules and user-level preventive maintenance procedures.

 Chapter 7, “Contact Information and Troubleshooting,“ provides

setup problems and signal alerts.

 Appendix A, “Acoustic Droplet Ejection Technology,” describes

acoustic droplet ejection technology.

xviii Echo Liquid Handler User Manual

 Appendix B, “Barcode Locations,” specifies where a microplate

barcode label must be placed to be read by the Echo liquid handler.

 Appendix C, “Chiller Information,”contains chiller descripton, setup

and operation, and troubleshooting.

Conventions used in this manual

Some of the text in this manual uses special formatting to help indicate emphasis or keystrokes. The text conventions are as follows:

Convention Example Meaning

Small caps, bold Press the

Courier client.txt

client.old2.txt

Blue, underlined Software installation

Quotation marks

See “Manage labware definitions” on page 5-31

NEXT button Indicates an on-screen button,

label, menu title, or menu item.

Indicates a display output, printed output, keyboard input, or file names.

Signifies a hyperlink to a topic.

Refers you to a topic elsewhere in the manual. Usually includes a hyperlink.

Preface xix

xx Echo Liquid Handler User Manual

CHAPTER 0INTRODUCTION

The Labcyte® Echo® 500 series includes the Echo 520, 550 and 555 liquid handlers.

Throughout this manual the descriptions, illustrations, and procedures apply to all liquid handlers unless the information is specifically stated for one model.

C HAPTER

All models utilize ADE (acoustic droplet ejection) technology patented by Labcyte Inc. Ultrasoundbased ADE precisely transfers nanoliter volumes of liquids, including DMSO, buffers with or without proteins or nucleic acids, surfactants, cell culture media and serum between microplates.

The Echo 500 series also utilizes Dynamic Fluid Analysis™, a patented Labcyte process that enables the liquid handler toadjust transfer parameters on the fly, without the need for user recalibration.

Excellent accuracy and precision in nanoliter volumes allow for direct transfer of compounds, which reduces the need for intermediate plates and multiple dilutions, and produces more accurate results in less time. The Echo liquid handlers dispense as little as 2.5 nL without using pins, tips, or washing.

Echo qualified source plates are specifically designed for the Echo liquid handler to take advantage of acoustic transfer technology and improve liquid transfer performance.

Nanoliter dispensing also makes an Echo liquid handler an excellent fit for many challenging life science applications in drug discovery, genomics, proteomics, and protein crystallography.

Figure 1.1 Echo 555 liquid handler

Introduction 1-1

1.1 Acoustic Droplet Ejection

Loading source microplate

Acoustic droplet ejection (ADE) technology is the process of transferring nanoliter volumes of liquid using acoustic energy. The Echo liquid handler uses this technology to transfer fluid droplets from a source microplate to a receiving surface.

The following is a simplified sequence of ADE events:

1. The Echo liquid handler creates an analog RF (radio frequency) wave.

2. The RF wave is amplified and transmitted to a focused acoustic transducer, which transforms the RF wave into an acoustic pulse.

3. This acoustic pulse is transmitted into the fluid in the source microplate well, causing the fluid to form a droplet that travels upwards.

4. The droplet is captured by the receiving surface, which may be an inverted destination microplate well or a flat substrate. Surface tension of the fluid keeps it on the receiving surface.

For more information about Acoustic Droplet Ejection Technology, see Appendix A

1.1.1 How does the Echo system work?

The Echo liquid handler uses the following basic steps to transfer liquid:

1. The robot (or operator) loads the source microplate into the Echo liquid handler. This step can also be performed by a robot.

The Echo liquid handler retracts the source microplate, then surveys and measures the fluid height in each well.

For aqueous solutions, the reported value options will vary by fluid type. For more details, see

Diagnostic Survey” on page 5-39.

“

For DMSO-based solutions, the software reports DMSO/water concentration (Echo 550 and Echo 555 only).

Note:

1-2 Echo Liquid Handler User Manual

2. The operator loads the destination plate (can be microplate or slide

Loading destination plate

Plate being inverted Destination plate down

holder) that will receive the liquid into the Echo liquid handler.

Before retracting the destination plate, the Echo liquid handler inverts the plate, with the open microplate wells or slide facing downward. This step is required to receive droplets ejected UP from the source microplate. See “

Will the fluid splash or fall out?” on page 1-4.

Figure 1.1 Inverting the destination plate

During the transfer process, the source microplate remains in a fixed position; the destination plate is positioned just above the source microplate and moves simultaneously with the acoustic transducer.

The acoustic transducer moves beneath the wells. At each well, the acoustic transducer emits acoustic energy that ejects a specified volume of liquid upwards to the receiving well or slide on the destination plate.

Figure 1.2 Diagram of transferring fluid

After liquid transfer, the Echo liquid handler moves the filled destination plate out of the instrument and inverts the plate again so that it can be removed.

3. The operator removes the destination plate containing the transferred compound.

Introduction 1-3

1.1.2 Will the fluid splash or fall out?

The Echo liquid handler performs liquid transfer with an inverted destination plate. Users might question whether all of the droplet remains in the destination plate well after transfer. Will any of it splash out? Will the repeated motions of the destination plate, or simply time passing, allow any of the transferred liquid to fall or drain out of the destination plate well?

Pre-loaded 1536-well destination microplates will not experience fluid loss when they are inverted. Pre-loaded 384-well destination microplates containing fluid with low surface tension may experience fluid loss when inverted and should be tested. Pre-loaded 96-well destination microplates may experience loss of fluid when inverted, depending on fluid type and volume, and are not recommended for “wet” (pre-loaded) fluid transfers.

1-4 Echo Liquid Handler User Manual

1.2 What are the Echo liquid handler features?

Echo liquid handlers are well suited to transferring low-nanoliter volumes of aqueous and DMSO-based solutions for a wide variety of applications. Echo liquid handler features that will be important to any organization include:

1.2.1 “Touchless” fluid transfers

Echo technology uses “touchless” fluid transfer.

 Nothing physically touches the fluid in the wells or the interior walls

of the microplate. Droplets are ejected using an ultrasound wave transmitted from below the source microplate. The ejected droplet is directly transferred upward from the supply plate to the receiver.

 No direct contact of the source fluid or the ejected droplet as it travels

to the destination plate, means no cross-contamination can occur.

“Touchless” fluid transfer provides the following benefits:

 Improved reliability

 Lower maintenance requirements

 Lower transfer costs

 Reduced waste

The following figure shows a single droplet ejected from a 384-well microplate. Droplets are discrete and of consistent size without the creation of spray or aerosol.

Figure 1.3 A single droplet ejection.

Introduction 1-5

1.2.2 Reliability

The Echo liquid handler has been designed for heavy duty-cycles and reliable operation. Compared to operating and maintaining existing liquid handling equipment, Echo technology offers the customer a simpler, trouble-free device.

Proactive control and monitoring of the systems reduce lost operational time or material if an error occurs:

 Multiple sensors monitor the coupling fluid system. One sensor

monitors the fluid level in the acoustic coupler catch basin. Another sensor monitors for fluid at the low point of the bottom pan.

 Compressed air and vacuum systems are monitored for correct supply

and operation.

 Each of the moving mechanisms includes multiple sensors that define

home positions and absolute limits, reducing the possibility of mechanical interference or misalignment.

When a problem is detected, the following actions occur:

 The coupling fluid pump immediately turns off (depending on the

problem).

 Front and rear panel fault lamps and LCD messages notify the

operator that a problem has occurred.

 ActiveX

events notify remote systems that are connected to the

Echo liquid handler.

1-6 Echo Liquid Handler User Manual

1.2.3 Ease of use

 Select a protocol

 Run the protocol

An Echo liquid handler is quickly ready for use.

 Minimal operator training is required.

 User calibration or recalibration are not required.

 Preparation for liquid transfers requires little more than defining a

transfer protocol and selecting the source and destination plates being used. Protocols or destination plates can be quickly defined by the operator.

 Liquid transfer requires only a few “clicks” by the operator, using the

software, which is included with each instrument.

 The Echo liquid handler can be operated manually by a researcher or

as part of a fully automated drug discovery lab.

For example, Figure 1.4 demonstrates the simple, two-step process required to run an existing fluid transfer protocol using the software program.

Figure 1.4 Executing a transfer protocol in two steps.

Introduction 1-7

1.2.4 System integration friendly

While manual mode (operator controlled via Labcyte software) and standalone mode (hand-loaded microplates) support all functionalities, the Echo liquid handler excels in its ease of integration into automated systems.

 The main body of the Echo liquid handler has a very small footprint:

53.9 cm wide and 68.3 cm deep (21.2 in x 26.9 in).

 Facilities requirements (power, temperature, ventilation, compressed

air, vacuum, communications) are simple and straight forward.

 Source and destination microplates are loaded onto stages presented

outside the instrument and placed in a fixed location for easy access by a robotic arm (see the following figure).

 Most barcode standards are supported. Barcodes can be located on

the short or long side of a microplate.

 The control software supports manual operation “out of the box.”

device control, system monitoring, creation of destination plates and protocol definitions, and protocol execution are available to the user.

 Customers choosing to develop their own Echo software applications

are supported by a fully implemented ActiveX control library.

Figure 1.5 Access™ workstation loading a destination microplate.

See “What types of liquid transfer are supported?” on page 1-12 for more

information.

1-8 Echo Liquid Handler User Manual

1.2.5 Low maintenance requirements

Echo instrument up-time is high, not only because it is reliable, but also because very little maintenance is required.

 Routine operator maintenance takes only minutes each week—

typically keeping the coupling fluid bottle clean and filled.

 Field service maintenance is scheduled once every six months.

 Echo technology means that there are no tips, no pins, and no

washing.

 The acoustic transducer itself has no moving parts to wear out. It

simply converts RF (radio frequency) energy to ultrasound energy. The mechanisms that move it into position are designed for a long and serviceable life.

1.2.6 Cost savings

The Echo liquid handler reduces operating costs by:

 Enabling the routine use of higher density, lower volume assays.

Moving to smaller assay volumes reduces the costs of compounds, solvents, and reagents.

 The use of higher density microplates. High density microplates

dramatically increase the number of assays that can be performed with a limited amount of test compound used.

 Minimizing the use of expendable supplies (for example, intermediate

microplates, tips, pins, and wash solutions).

 Eliminating washing stations and their complex procedures and fluids.

 Eliminating the requirement to calibrate reduces upkeep time or

delays in adopting new liquid handling procedures.

Introduction 1-9

1.2.7 High accuracy, precision, and speed

High throughput capabilities are maintained without degrading fluid transfer accuracy and precision.

Droplet-to-droplet volumetric precision exhibits a low CV (coefficient of variation) for all fluid types tested to date. This is attributed to the elimination of sample contact with the ejection mechanism. In addition, dynamic real-time measurement of the fluid at multiple times during a run allows the system to adjust automatically and instantaneously to changes in fluid behavior.The use of a single, high-speed, serial mechanism reduces sample-to-sample variability with the Echo liquid handler as compared to tip-based and other instruments that use multiple mechanisms in parallel.

1-10 Echo Liquid Handler User Manual

1.2.8 Value-added quality control

In addition to precise and accurate transfer of fluids, the Echo liquid handler has an auditing function that can provide quality control of fluid samples. The Echo liquid handler applies a low-energy sound pulse to the fluid sample in each well, and the reflected signal is used to calculate the following:

 DMSO-based solutions: fill height and DMSO/water concentration.

 Glycerol-based solutions: percentage of glycerol concentration

(up to 50%).

 Protein crystallography fluids: impedance measurement in

megarayls (unit of acoustic impedance). In approximate values, solutions at low megarayl values require less energy to transfer.

The data can then be uploaded to a LIMS (Laboratory Information Management System) for post transfer processing and analysis.

For example, the following figure shows part of the plate survey of samples in CP buffer. The plate survey was saved to CSV (comma separated values) file format and opened in Microsoft® Excel®.

Figure 1.6 Example of aqueous fluid thickness data imported into a spreadsheet for analysis.

Introduction 1-11

1.3 What types of liquid transfer are supported?

The Echo liquid handler supports a variety of simple to complex microplate liquid transfer protocols. The bundled software program allows a user to set up the following types of protocol transfers:

Table 1.1 Liquid transfer types

Source (Src)

Microplate

384-well 96-well

384-well 384-well standard (1 src plate to 1 dest plate)

384-well 1536-well

384-well 3456-well

1536-well 96-well

1536-well 384-well

1536-well 1536-well standard (1 src plate to 1 dest plate)

1 Destacking: transferring liquid from a single source (src) plate to several

destination (dest) plates

2 Stacking: transferring liquid from several source plates to a single destination

plate

Destination

(Dest)

Microplate

Transfer map type

(full or partial)

destacking

quadrant (4 regions from 1 src plate to 4 dest plates)

stacking

quadrant (4 src plates to 4 regions on 1 dest plate)

stacking quadrant (9 src plates to 9 regions on 1 dest plate)

destacking quadrant (16 regions from 1 src plate to 16 dest plates)

destacking quadrant (4 regions from 1 src plate to 4 dest plates)

destacking interleaved (2x2, or every other dest well)

stacking interleaved (2x2, or every other dest well)

stacking interleaved (3x3, or every third dest well)

destacking interleaved (4x4, or every fourth dest well)

destacking interleaved (2x2, or every other dest well)

Liquid transfer can be tailored to the user’s specific requirements. Variables that are user selectable or definable include the following:

 Source microplate type (from the list of qualified plates)

 User-definable destination microplate or surface

 Transfer type (for example, interleaved or quadrant 384-well to

96-well transfers)

 Transfer volume

 Number of copies (how many times the transfer protocol is executed)

 Transfer map specifying exactly which wells, or regions of wells, will

be transferred

 Predefined DMSO/water concentration maps

These fluid transfer attributes and choices are defined in Chapter 5.

1-12 Echo Liquid Handler User Manual

Examples of these protocols are described later in this section.

A1 A1

Additional and more complex fluid transfer protocols can be defined with the Echo Applications software packages (Echo Plate Reformat, Echo Cherry Pick, Echo Dose-Response, and Echo Array Maker), as well as with the extensive ActiveX control library defined in the Echo Software Development Kit (Echo SDK). These protocols can specify virtually any combination of transfers from source microplate wells to destination wells and slides. Refer to the Echo Application manuals and Echo Integration Guide for more information.

1.3.1 384-plate to 384-plate liquid transfer

Figure 1.7 and Figure 1.8 below illustrate simple fluid transfers between two 384-well microplates. While Figure 1.7 shows a full-plate transfer, Figure 1.8 shows a multi-region, partial plate transfer. Both transfer protocols are easily defined by the operator using the Echo liquid handler software.

Figure 1.7 384 to 384 full plate transfer

Figure 1.8 384 to 384 multi-region, partial plate transfer.

Introduction 1-13

Figure 1.9 illustrates a more complex fluid transfer between two 384-well microplates. Droplets are ejected from multiple source plate wells into the same destination plate well (pooling). This transfer map and its transfer protocol cannot be defined using the current software program, but it can be defined using the Echo Applications software or with the Echo ActiveX programming interface (refer to Echo Integration Guide).

Figure 1.9 384 to 384 multiple source plate wells pooled into one destination plate well transfer.

1.3.2 384-plate to 96-plate liquid transfer

Figure 1.10 illustrates a 384-well to 96-well quadrant fluid transfer.

Wells from a single 384-well plate can be transferred, or destacked, to four 96-well plates. Individual wells or all wells from each quadrant can be transferred to the corresponding wells in the destination plate.

Figure 1.10 384 to 96 plate transfer.

1-14 Echo Liquid Handler User Manual

1.3.3 384-plate to 1536-plate liquid transfer

Figure 1.11 illustrates a 384-well to 1536-well interleaved fluid transfer. An interleaved transfer places, or stacks, all source A1 well transfers into a single cluster on the destination plate. Figures 1.12 and 1.11 clearly show these two schemes.

Each source microplate requires its own transfer map defining which wells are to be transferred to the destination microplate. These transfer maps can be unique for each of the source plates.

If the transfer maps consist of single- or multi-region transfers, the transfer protocol can be easily defined using the software.

Complex transfer maps can be created using Echo Applications software or ActiveX programming interface in the Echo Software Development Kit (refer to the Echo Integration Guide).

Figure 1.11 384 to 1536 interleaved full plate transfer.

Introduction 1-15

1.3.4 1536-plate to 384-plate liquid transfer

Figure 1.12 illustrates a 1536-well to 384-well quadrant fluid transfer.

Wells from a single 1536-well plate can be transferred, or destacked, to four 384-well plates. Individual wells or all wells from each quadrant can be transferred to the corresponding wells in the destination plate.

Figure 1.12 1536 to 384 quadrant full plate transfer

1-16 Echo Liquid Handler User Manual

1.3.5 1536-plate to 1536-plate liquid transfer

Fluid transfers between two 1536-well microplates are exactly the same as 384-plate to 384-plate transfers, except for the well densities.

Note: Echo 555 only: High-throughput transfer of DMSO is available for the 1536-well LDV plate (1536LDV_DMSO_HT). This special calibration type uses the same 1536-well LDV plate that is used for standard DMSO and aqueous transfer; however, the repetition rate for droplet transfer is much faster. This calibration type is ideal for preparing multiple screening plates and prewetted plates.

Note: For liquid transfer of very small volumes, or transfer that requires precise droplet placement, use the 1536LDV_DMSO calibration.

Introduction 1-17

1.4 What system configurations are supported?

The Echo liquid handler can be used in a manual configuration or in several levels of automated, system integrated configurations. This section explores the following operating modes:

 Manual mode  Software-driven mode  Modular workstation mode  Small cell mode  Fully automated mode

The server/client architecture allows multiple client computers/programs to connect to an Echo liquid handler at the same time. Read “

considerations” on page 1-23 for a discussion of the advantages and

disadvantages of multiple-client operation.

Note: The software comes with a static IP address (192.168.1.1). If this IP address is not available, or if you are installing more than one Echo liquid handler on a network, use the Echo Admin Client (on the Echo Server 2.4 disk) to change the IP address.

Multi-client

1.4.1 Manual operation

Manual operation leaves control of every step to the operator:

 Defining new fluid transfer protocols or new destination microplates  Executing a fluid transfer protocol  Loading and unloading both source and destination microplates  Monitoring status

Figure 1.13 Manual operation.

Manual operation is also used when performing the following functions:

 Defining new destination microplates  Defining new fluid transfer protocols  Performing periodic maintenance and calibration

For more information about manual operation, read Chapter 4.

1-18 Echo Liquid Handler User Manual

1.4.2 Echo applications

Labcyte provides the following Echo Applications that extend beyond the basic Echo software to enable the operator to easily create a variety of more complex destination plates.Labcyte currently provides the following software applications:

 Echo Dose-Response: eliminates the tedious and error-prone steps

in manual serial dilution to produce assay-ready microplates for IC50, EC50, and dose-response curves; includes creation of intermediate plate for additional concentrations; generates customizable reports in common data formats.

 Echo Cherry Pick: transfers sample from any source well to any

destination well; uses customer-provided pick list to automate platemapping; generates customizable reports for tracking sample data.

 Echo Plate Reformat: automates microplate replication, down-

stacking and up-stacking microplates; also provides custom plate mapping for pooling, transposing or interleaving samples.

 Echo Array Maker: automates sample transfer for specialized

microplates, such as protein crystallization; enables custom array creation to SBS-compliant destination labware.

 Echo Plate Audit: enables customers to collect information on

sample properties such as fluid height, fluid volume, and composition; quickly assess sample quality across plates.

For customers requiring additional customization, Labcyte also provides Echo application programming interfaces and code examples to integrate the Echo system into their own laboratory automation system.

Contact your Labcyte representative for information on the Echo applications listed above or on the Echo SDK (software development kit).

Introduction 1-19

1.4.3 Modular workstation operation

Modular workstation operation differs from manual operation with the addition of a robot to automate the loading and unloading of source and destination plates. Typically, a custom program is written that controls and synchronizes the Echo liquid handler and robot interactions.

An operator is still involved and performs the following functions:

 Deliver master source microplates from the compound library.

 Maintain a supply of empty destination microplates.

 Select and execute the fluid transfer protocol.

 Deliver the filled assay microplates to the next step in the process.

 Monitor the status of the Echo liquid handler and the robot, possibly

with the assistance of a custom software program.

Figure 1.14 Modular workstation operation

1-20 Echo Liquid Handler User Manual

1.4.4 Small cell operation

Small cell operation includes the automation of HTS processes other than liquid transfer. Those processes might include several of the following:

 Lidding and delidding  Microplate labeling  Microplate sealing  Incubation  Reaction detection  Temperature or humidity control  Reagent addition  Wash station (used with other instruments)  Small cell mode operation

Figure 1.15 Small cell mode operation

Introduction 1-21

1.4.5 Fully automated operation

A more fully automated operation expands on the small cell concept and may incorporate a larger range of processes and operations.

Figure 1.16 Full automated mode operation

Larger, fully automated drug discovery labs might include the following:

• A larger combination of the processes and stations listed in the discussion of a small cell mode operation.

See “Small cell operation” on page 1-21.

• One or more supervisory workstations.

• A conveyor system to move microplates and other material not only between stations, but into and out of the lab.

• Multiple robots, individually chosen and programmed to support a specific function or machine.

• Multiple liquid handlers.

Note: Echo Admin Client allows the operator to change the IP address on the client. This is included with the Echo Server

2.4 disk.

1-22 Echo Liquid Handler User Manual

1.4.6 Multi-client considerations

Larger research labs may have many computers, instruments, controllers, and other devices interconnected on an Ethernet LAN (local area network). Please review the following information and take appropriate measures to avoid conflicts caused by multiple workstations controlling an Echo liquid handler inappropriately.

The Echo liquid handler can be connected to a network or to more than one workstation. This configuration allows one or more users to control the Echo remotely, to download liquid transfer information directly to the network, or to connect more than one Echo liquid handler to an automated filling operation.

Figure 1.17 Multiple clients talking to an Echo liquid handler.

For example, a local workstation might be used to do the following:

 Monitor instrument status.

 Perform user maintenance.

 Execute troubleshooting procedures.

At the same time, a central system may be used to control all of the day-today work flow involving the instruments in the lab. In relation to an Echo liquid handler, it might be involved in the following:

 Define fluid transfer protocols.

 Define destination microplates.

 Deliver source and destination microplates to the Echo liquid handler

via conveyor belt and robot.

 Remove source and destination microplates.

 Execute fluid transfer protocols.

 Handle all of the data.

Introduction 1-23

The Echo liquid handler is a client/server system. The “client” is the external workstation that contains the user interface software to define plate and protocol information. The “server” is the controller that runs the instrument motors and sensors. Each subsystem is independent, with its own system checks. This architecture allows the Echo liquid handler to be commanded by multiple clients to run concurrent operations, yet ensure that each operation is “safe” with respect to each other.

For example, one user might command the Echo liquid handler to do a plate survey, then a liquid transfer protocol. During the source plate survey, another user might command the Echo liquid handler to extend the source plate. The plate survey that is in progress will continue to completion, then extend the source plate. Thus, while the Echo liquid handler guarantees "safe" operation, it cannot prevent the commands of one user interfering with those of a second user.Therefore, it is the responsibility of the users to coordinate their actions to ensure the proper fulfillment of a transfer protocol.

Also, if more than one user creates, edits, or deletes fluid transfer protocol definitions or destination microplate definitions, then there must be communication between users to ensure that the definitions are consistent with the protocols.

Users should be aware of the following considerations with the client/ server architecture.

 An Echo liquid handler is not limited to a single client. One or more

clients can either monitor its status or command it to execute its functions.

 Uncoordinated clients could cause out-of-order execution that

interferes with other users' protocols. For example, client “A” could delete or edit a protocol or destination microplate definition created by client “B.” An edited protocol definition would only be detected if the original definition was stored on client “B” and compared to the definition in the database.

1-24 Echo Liquid Handler User Manual

C HAPTER

CHAPTER 0INSTALLATION AND RELOCATION

Labcyte installs the Echo liquid handler and Echo software for the customer. However, site preparation and additional software installation are the customer’s responsibility.

This chapter covers pre-installation requirements, additional software installation, and instrument relocation.

This chapter also provides a brief overview of the shipment contents and installation steps performed by Labcyte field engineers.

 Pre-installation requirements

 Installation overview

 Software installation

 Instrument Relocation

Installation and Relocation 2-1

2.1 Pre-installation requirements

(

)

(

)

118 cm (46.5 in)

Operating

Side clearance

2.5 cm (1 in)

30.5 cm (12 in)

Rear clearance

Top clearance

25.4 cm (10 in)

Envelope

7.6 cm (3 in)

Front clearance

2.1.1 Site requirements

The following site requirements must be met before installation:

 Operating environment of 19°–26°C (66°–79°F) and 10%–80%

relative humidity, non-condensing.

 Sturdy, level, water-resistant bench top capable of supporting

128 kg (283 lbs) with sufficient bench space to accommodate the Echo liquid handler in a minimum operating envelope as follows (see Figure 2.1):

Operating space envelope: 59 cm (23.2 in) width x 99 cm (39 in) depth x 118 cm (46.5 in) height

Figure 2.1 Instrument layout on bench top

2-2 Echo Liquid Handler User Manual

Top clearance: 25.4 cm (10 in) minimum above the instrument to

remove the top cover for servicing.

Note: Less height clearance (>2.5 in) is acceptable if there is sufficient space to lift the top and slide it all the way forward. Talk to your Labcyte field engineer.

Front clearance: sufficient clearance to load and unload microplates by hand or by robot.

Side clearance: 2.5 cm (1 in) minimum on left and right sides.

Rear clearance: 30.5 (12 in) minimum behind the instrument to

allow for cables, hoses, tubing, and maintenance access

Note: The coupling fluid bottle and caddy fit within the rear clearance. If you position the bottle and caddy on the side of the instrument, add at least 17.8 cm (7 in) to the side clearance. The caddy must be placed within 16” of tubing run to the Echo.

Additional space for the client PC, coupling fluid chiller (if it is on the bench), and microplates. Additional space would also be needed if you use robotics.

 AC power: The user must provide AC power matching one of the

two following power configurations:

 115 VAC, 50/60 Hz, 10 A  230 VAC, 50/60 Hz, 5 A

Note: The power configuration is factory set and cannot be changed onsite.

 Compressed air source: Clean, dry air at 552 kPa (80 PSI) minimum,

1034 kPa (150 PSI) maximum. Consumption not to exceed 1cf/m @ 150 PSI.

Labcyte provides the compressed air line and fitting with the Echo liquid handler, but the customer must provide a source compatible with 170 in ID/.250 in OD tubing.

 Vacuum source: Vacuum returns the coupling fluid to the supply

bottle and dries source plates before they are unloaded. Customer must provide a fitting that connects 3/8”ID, 5/8”OD tubing to either house vacuum or vacuum pump. Vacuum source should have the following specifications:

House vacuum: Minimum vacuum at Echo connection: 200 Torr (266 mbar, 22 in Hg Vac)

Vacuum system capacity steady state: 15 L/min (0.9 m3/h, 0.53 cf/m)

Drying surge: 80 L/min for 10 sec, (4.8 m

/h, 2.8 cf/m),

33% max duty cycle.

Vacuum pump: Minimum vacuum at Echo connection: 100 Torr (133 mbar, 26 in Hg Vac)

Pump flow rate: with 10 L surge tank: 40 L/min (2.4 m3/h, 1.4 cf/m) without surge tank (not recommended): 80 L/min (4.8 m

/h, 2.8 cf/m)

Note: The 10 L surge tank comes standard with the Echo liquid handler to ensure consistent vacuum.

Installation and Relocation 2-3

2.1.2 Additional components

The following components are needed to work with the Echo liquid handler:

 Chiller for maintaining a constant fluid temperature. Included with

the Echo system. See “ information.

 Distilled water for filling the chiller well, coupling fluid bottle, and

coupling fluid circuit.

Caution: Do not use de-ionized water, as it may corrode the fluidics

system.

 Client PC (customer’s computer) running the Echo software to

monitor and control the Echo liquid handler. The PC should have an Intel® Core™ 2 Duo processor or equivalent with Windows XP Professional (SP3 or later); 1GB of RAM, minimum; 1 GB of available disk storage; video resolution of 1280 x 1024; and a 10/100/1000 baseT Ethernet port.

Note: If you are integrating the Echo liquid handler into an automated HTS (High Throughput Screening) system and need to install the Client + Server software onto a workstation, it must contain an Intel Core™ 2 Duo processor or better.

Chiller Information” on page C-1 for more

 Network IP address: A static IP address must be allocated for the

Echo liquid handler. If you cannot use the preconfigured IP address (192.168.1.1) in your network, or if you are installing more than one Echo liquid handler, use the Echo Admin Client (included on the Echo Server 2.4 disk) to change the IP address on the client PC.

2-4 Echo Liquid Handler User Manual

2.2 Installation overview

Ethernet cable

Compressed air line

Vacuum line

Client PC

Chiller

Echo liquid handler

fluid lines

Chiller

Bench

Floor

(cross-over)

Surge tank

The Labcyte Echo liquid handler is installed by Labcyte field engineers. The following is a brief overview of the hardware and software installation steps they perform:

1. Unpack the Echo liquid handler and place it in the area that meets the minimum operating space requirement.

2. Connect the Echo liquid handler to AC power, air pressure, vacuum pressure, surge tank, coupling fluid bottle, and the coupling fluid chiller.

3. Fill the coupling fluid bottle and prime the coupling fluid circuit.

4. Perform a basic functional check of the Echo liquid handler.

5. Perform the Site Acceptance Test (SAT) process (optional).

6. Change the Echo IP address, if a different IP address is required by your

system administrator.

7. Install the Echo software onto the customer-supplied PC (client workstation).

8. Connect the client PC to the Echo liquid handler and verify communications.

Figure 2.2 Installed system

Installation and Relocation 2-5

2.3 Software installation

Labcyte installs the Echo software onto the client PC that is connected to the Echo liquid handler. If you need to install the software onto additional computers, such as network PCs, use the instructions provided in this section.

See “

Multi-client considerations” on page 1-23 for advantages and

disadvantages of connecting more than one workstation.

Note: If you are connecting the Echo liquid handler to a LAN via a hub, switch, or router, use a standard ethernet cable. If you are connecting the Echo liquid handler directly to a workstation, use the included “cross-over” Ethernet cable.

During installation the Echo software will provide the following installation types:

 Client (Only): Installs the Echo software onto the client PC that is

connected to the Echo liquid handler (normal mode). The software allows the operator to communicate with, control, and monitor an Echo liquid handler through easy-to-use software screens (Graphical User Interface). This option requires a host name or IP address of the Echo server.

 Client and Server:

 Normal Mode installs the Echo client software onto the client PC

and the Echo server software onto the Echo controller. This software is installed by Labcyte field engineers.

 Stub Mode creates a virtual Echo instrument onto the client PC

that developers can use to develop and test custom software.

Note: Use this installation type only if you are writing your own programs that will call the Echo ActiveX programming interface. Call Labcyte Service and Support if you have any questions regarding this installation type.

1 Ethernet cable that switches, or crosses over, receive and transmit wire pairs that enable

two devices to communicate with each other.

2-6 Echo Liquid Handler User Manual

2.3.1 Install the software

The following procedure describes installation of Client (Only) software. For information on installing Client + Server software, refer to the Labcyte Echo Integration Guide.

To install Echo liquid handler software:

1. Insert the Echo liquid handler software installation CD into the CD drive. The installation should automatically start.

If the installation does not start:

a. Go to My Computer and double-click the drive icon for the CD drive.

b. Look for setup.exe in the directory on the CD and double-click the

icon.

c. Select Install Client (Only)

2. Enter the Host Name or IP address that will be used by the Echo server. The Echo liquid handler comes pre-configured with IP address

192.168.1.1 and a subnet mask of 255.255.0.0. If you cannot use this IP address, run the Echo Admin Client on the Echo Server 2.4 disk to enter a different IP address.

Figure 2.3 Entering host name

Installation and Relocation 2-7

3. Click N

EXT and and then INSTALL to start the installation. Edit or verify

the previously entered information or selections by clicking the BACK button.

Figure 2.4 Starting installation

A dialog window will report the installation’s activity and progress.

4. Click the FINISH button when the installation is done.

Note: Read the release notes for information about the version of

software being installed. The release notes are stored in the Labcyte Echo liquid handler folder on your Windows Desktop for later viewing.

2-8 Echo Liquid Handler User Manual

2.3.2 Uninstall the Echo software

If you need to uninstall the Echo software, use the following procedure:

1. Click the START button on the Windows Taskbar.

2. Point to SETTINGS and select CONTROL PANEL.

3. Double-click ADD OR REMOVE PROGRAMS.

4. Select Echo liquid handler and click REMOVE.

2.3.3 Upgrade the Echo software

Upgrading the software requires a new Echo software CD. Contact Labcyte Service and Support for the most current version. See “Contact

information” on page 7-1.

Depending on your current software version, you may need to run a clean installation. Check with Labcyte Service and Support to determine if your software can be upgraded. If not, you will need to perform a new (or clean) installation. Back up your existing liquid transfer and plate definition data.

1. Insert the new Echo liquid handler software CD on the client PC.

2. Follow the installation instructions. See “Install the software” on

page 2-7.

Installation and Relocation 2-9

2.4 Instrument Relocation

Note: If you do not plan to set up the Echo liquid handler within two

weeks, refer to “

2.4.1 Prepare the Echo liquid handler for a move

Use the following procedure if the Echo liquid handler must be moved:

1. Determine if the move must involve Labcyte.

 If the move requires transportation (truck, air, train) call Labcyte

Service and Support. Do not attempt to move the Echo liquid handler yourself.

 If the move can be easily accomplished using a moving dolly or cart,

the move can be performed without Labcyte involvement. If you have any questions, contact Labcyte Service and Support See “

information” on page 7-1.

2. Finish any operation in progress.

3. Remove microplates and microplate inserts from the Echo liquid handler.

Extended Non-Use and Storage” on page 6-19.

Contact

4. Turn off the Echo liquid handler and the fluid chiller.

See “Turn off Echo power” on page 4-5 and “Chiller Setup and Operation” on page C-3.

5. After the Echo liquid handler has been powered down, disconnect the following lines from the back panel.

 Chiller input and output tubing  Compressed air supply line  Vacuum supply line  Ethernet cable  AC power cable  Coupling fluid bottle, caddy, and cable

6. Use proper safety procedures to lift the Echo liquid handler gently onto a moving dolly or cart of sufficient size and strength to support the unit.

Warning: The Echo liquid handler weighs approximately 128 kg (283 lb.). Moving or lifting the Echo liquid handler incorrectly can cause severe injury. Never attempt to move or lift the Echo liquid handler without using proper equipment and trained personnel.

7. Move the Echo liquid handler and ancillary equipment and material (bottles, tubing, microplates, source plate inserts, etc.) to the new location, which should already be prepared and comply with all preinstallation requirements.

See “

Pre-installation requirements” on page 2-2.

2-10 Echo Liquid Handler User Manual

2.4.2 Reinstall the Echo liquid handler after a

Fluid level sensor

move

After the move, use the following procedure to restore an Echo liquid handler to normal operation.

1. Ensure that the new mounting surface is level.

2. Set up the Echo liquid handler, bottles, computer, and other ancillaries.

Remember to allow for minimum operating space around the Echo liquid handler. See “Site requirements” on page 2-2.

3. Set up and start the chiller. See “Chiller Setup and Operation”in Appendix C for instructions.

4. Set up the coupling fluid bottle. Ensure that the bottle cap is firmly attached to ensure a tight vacuum seal. Place the filled coupling fluid bottle in the caddy. The fluid level sensor (red LED) should be visible on the side wall of the instrument to confirm that the bottle is positioned correctly and is filled.

Figure 2.5 Fluid level sensor in bottle caddy

Note: Older systems hold the fluid bottle in a separate caddy with the sensor showing through the hole on the top of the caddy. Also, some older systems use a waste bottle.

Installation and Relocation 2-11

5. Connect the air pressure and vacuum pressure supply lines to the fluidics

Coupler fluid bottle cap

(blue bottle cap)

From vacuum source

To coupler catch nozzle

(long, straight tube)

From catch basin

(J-tube)

From chiller (WATER OUT)

To chiller (

WATER IN)

panel. The connections are labeled for easy reference.

Figure 2.6 Bottle connections

Caution: Do not run the pump until the vacuum source is connected

and active. Running the pump without vacuum can lead to flooding the system or a system error.

6. Connect the chiller. Attach the plastic tubing between the chiller and the Echo liquid handler.

Figure 2.7 Tubing connections on the chiller

2-12 Echo Liquid Handler User Manual

Caution: Do not run the pump until the chiller tubing is connected.

Fluidics panel

From chiller (Water In)

To chiller (Water Out)

Compressed air line (Air In)

Vacuum line (Vacuum)

Running the pump without the chiller tubing may damage other system components.

Figure 2.8 Tubing connections on the Echo system

7. Use the "cross-over" Ethernet cable to connect the Echo liquid handler to the client PC. The client PC should have the Echo liquid handler software already installed.

Fill the coupling fluid bottle and the fluidics circuit. See “Refill the coupling

fluid bottle” on page 6-8 and “Cycle fluid through the Echo system” on page 6-13.

Installation and Relocation 2-13

2-14 Echo Liquid Handler User Manual

C HAPTER

CHAPTER 0SYSTEM DESCRIPTION

This chapter covers the following Labcyte Echo liquid handler features and components:

 System overview

 Front panel

 Back panel

 Component description

 Process door

 Source plate gripper stage  Source plate insert  Destination plate gripper stage  Status indicator lights  LCD screen  Emergency Off switch  Anti-static bars  AC power and fuse compartment  Coupling fluid bottle  Fluid chiller  Surge tank  Fluidics panel

 Specifications

 Physical

 Mechanical  Environmental  Electrical  Communications  Fluid transfer  Client PC  Supported labware  Coupling fluid

System Description 3-1

3.1 System overview

Source plate gripper stage with plate insert

Destination plate gripper stage

Status indicators

LCD screen

Emergency off switch

Upper anti-static bar (behind front cover)

Lower anti-static bar (behind front cover)

Process door (shown open)

Barcode scanner, optional (behind front

This section provides the location and brief description of the components, followed by detailed descriptions.

3.1.1 Front panel

Figure 3.1 Echo Liquid Handler front panel

3-2 Echo Liquid Handler User Manual

Process door: The process door protects the operator from the moving

mechanisms blocks stray light emitted by the barcode scanner lasers. For more information, see “

Process door” on page 3-5.

Destination plate gripper stage: The destination plate gripper stage is the upper plate stage that extends from the Echo liquid handler. The plate gripper (end portion of the stage) holds the destination microplate that will receive the liquid being transferred). For more information, see

“

Destination plate gripper stage” on page 3-8.

Source plate gripper stage: The source plate gripper stage is the lower plate stage that extends from the Echo liquid handler. The plate gripper (end portion of the stage) holds the source microplate containing the liquid that will be transferred. For more information, see “Source plate gripper

stage” on page 3-6.

Source plate insert: The plate insert fits between the source plate and plate gripper to ensure that the source plate is properly positioned. For more information and table of insert types, see “

Source plate insert” on

page 3-6.

Figure 3.2 Plate inserts

Status indicator lights: There are three status indicator lights on the front of the Echo liquid handler:

 Power (green): The Echo liquid handler is on and ready.

 Warning (yellow): A component is out of position or a parameter is

out of range and requires attention.

 Fault (red): A problem has occurred that stops the current task and

requires immediate attention.

These status indicator lights are also provided on the back panel. For more information, see “Status indicator lights” on page 3-9.

LCD screen: The LCD screen on the front of the Echo liquid handler reports what the Echo liquid handler is doing or what operator action is required. This display is also provided on the back panel. For more information, see “LCD screen” on page 3-11.

Emergency off: The EMO (E

mergency Motion Off) switch shuts down all mechanical Echo liquid handler activity. This switch is also available on the rear panel.

Upper and lower anti-static bars: The upper and lower anti-static bars remove any electrostatic charge from the source and destination microplates. Electrostatic charge can affect the travel of the droplet. For more information, see “

Anti-static bars” on page 3-12.

Barcode scanner option: An Echo liquid handler can be purchased with one or more barcode scanners that will read and report barcoded microplates. For more information, see “Barcode scanner” on page 3-12.

System Description 3-3

3.1.2 Back panel

Status lights LCD

screen

Emergency Off (EMO) switch

AC power and fuse

compartment

Tubing and hose connections

Data connection

Coupling fluid bottle and caddy

Remote Emergency Off cable

Figure 3.3 Echo liquid handler back panel

AC power and fuse compartment: The AC power and fuse compartment (also called the power input module) contains the main Echo liquid handler power on/off switch, AC power outlet, and fuse compartment. For more information, see “AC power and fuse

compartment” on page 3-14.

Coupling fluid bottle and caddy: The 1000 mL coupling fluid bottle provides the coupling fluid (distilled water) that is pumped through the acoustic transducer to maintain constant temperature and transfer acoustic energy to the well. The coupling fluid bottle caddy contains a sensor that monitors the fluid level and issues a warning when the fluid level is less than 250 mL. For more information, see “

Coupling fluid bottle” on

page 3-15.

Status light indicators, LCD screen, and Emergency Off (EMO): Same as the components on the front panel. Remote EMO cable enables the customer to set up a remote EMO button more accessible to the operator.

Data connection: RJ-45 Ethernet network port connects the Echo liquid handler to a PC or to a network. For more information, see “

connection” on page 3-14.

Tubing and hose connections: the Echo liquid handler uses tubing and hoses to connect to the chiller, house air, surge tank, and vacuum pump.

Data

Waste water bottle (not shown, on older Echo systems only): The 500 mL waste water bottle collects overflow from the catch basin.

3-4 Echo Liquid Handler User Manual

3.2 Component description

3.2.1 Process door

The process door is normally closed to protect the operator from moving mechanisms in the Echo liquid handler and from any stray light emitted by the barcode scanner lasers. When a fluid transfer protocol is executed, the process door will be opened automatically, allowing easy human or robotic loading or unloading of microplates.

Figure 3.4 The process door (closed)

Warning: Any time the process door is open, whether or not one of the microplate assemblies has been extended outside of the Echo liquid handler, there is a pinch hazard. The pinch point label shown here is visible on both microplate assemblies and reminds you to keep your hands and fingers clear of the mechanisms.

System Description 3-5

3.2.2 Source plate gripper stage

Source plate gripper stage

The source plate gripper stage is the lower plate stage that extends from the Echo liquid handler. The plate gripper holds the microplate containing the liquid that will be transferred.

Note in Figure 3.5 that the source plate gripper stage extends out of the very bottom of the process door.

Figure 3.5 The extended source plate gripper stage.

3.2.3 Source plate insert

To keep the source plate secure, the plate gripper includes a plate insert designed for specific plate types. See the following table of plate inserts.

Table 3.1 Labcyte plate inserts

Plate insert Plate catalog # Insert size

384-well polypropylene P-05525 2.10 mm

1536-well COC high base LP-03730 4.50 mm

384-well LDV COC LP-0200 4.50 mm

1536-well LDV COC LP-0400 4.50 mm

Note: Handle the plate inserts carefully. They can be damaged from dropping or rough handling, which can affect instrument performance.

3-6 Echo Liquid Handler User Manual

When you are changing plate inserts, look for

Pads

Pusher plate

Sensors

Insert sensor

Plate gripper

Source plate insert

assembly

the insert size, which is located in the upper right corner.

When you place the microplate insert onto the plate gripper, the sensors will be flush against the insert sensor. The plate insert will rest on top of three pads and should move easily from side to side. See Figure 3.6.

Figure 3.6 Source plate gripper stage (entire assembly)

The plate insert will be moved into the correct position by the gripper arm when the source plate gripper stage moves through the process door.

When you load a source microplate onto the plate insert, well A1 is located in the upper left corner (see Figure 3.7). As a handy reminder, the upper left corner of the plate insert is labeled A1. The insert contains sensors that inform the Echo liquid handler which plate is being used.

System Description 3-7

Figure 3.7 Plate insert and gripper arm securely hold the source microplate.

A1 well location

Gripper arm

Plate insert

Sensors

A1 well location

3.2.4 Destination plate gripper stage

The destination plate gripper stage is the upper plate stage that extends from the Echo liquid handler. The plate gripper holds the microplate that will receive the liquid being transferred.

When you load the plate onto the plate gripper, well A1 goes into the upper left corner (nearest the Echo model number).

Figure 3.8 The extended destination plate gripper stage.

When the destination plate gripper stage is commanded to retract into the Echo liquid handler, the microplate is gripped and simultaneously rotated down. It is in this inverted position that the microplate will be able to receive the fluid droplets ejected upwards from the source plate wells.

3-8 Echo Liquid Handler User Manual

Figure 3.9 Inverting the destination plate

Destination microplate up Destination microplate downPlate being inverted

Power: Green = Ready

Warning: Clear = OK;

Yellow = Warning

Fault: Clear = OK;

Red = Fault

As the destination plate gripper stage retracts into the Echo liquid handler, a sensor detects whether a plate is or is not present. Normally, a transfer will not be executed if a destination plate is not detected. The human operator, or a controlling program, can override the plate presence check, if necessary.

When the destination plate gripper stage extends out of the Echo liquid handler, the plate is rotated back to its upright position.

3.2.5 Status indicator lights

There are three status indicator lights on the front of the Echo liquid handler and a duplicate set on the rear panel.

A quick glance can confirm whether the Echo liquid handler is operating normally or that it needs operator attention. The Power light (green) is on when the instrument on and initialized; the Warning light (yellow) turns on when a condition is outside of normal limits, but the instrument can continue to operate; the Fault light (red) turns on when a condition exceeds warning conditions and the instrument can no longer operate.

Figure 3.10 Echo liquid handler front panel

Power light

 Green: Normally the Power/Ready light is turned on, indicating that

the Echo liquid handler is both turned on and that its internal controller has completed its initialization and is ready.

 Off: When the Power/Ready light is turned off, either the Echo liquid

handler has not been turned on or its internal controller has not completed initialization.

System Description 3-9

Warning light

 Off: Normally the Warning light is turned off, indicating that the Echo

liquid handler is functioning correctly.

 Yellow: The Warning light is turned on when any of the following

conditions are true:

 The Echo liquid handler stages are not homed or are in the process

of being homed. See “Home calibration” on page 5-45.

 The coupling fluid bottle level is low. See “Coupling fluid

indicator” on page 5-35.

Note: When the coupling fluid falls below its recommended minimum level, air bubbles may be introduced into the coupling fluid, which reduces the accuracy or precision of source plate surveys and fluid transfers. Labcyte recommends that you refill the coupling fluid bottle as soon as possible after the Warning light has been turned on (see “Refill the coupling fluid bottle” on

page 6-8).

 The coupling fluid temperature is outside of the normal range:

22.1°C + 0.9°C (21.2°C – 23.0°C)

 The process door is neither fully open nor fully closed.  The upper anti-static bar is neither fully up nor fully down.  The source plate dryer nozzle is neither fully up nor fully down.

Fault light

 Off: Normally the Fault light is turned off, indicating that the Echo

liquid handler is functioning correctly.

 Red: The Fault light is turned on when any of the following

conditions are true:

 The user-supplied compressed air supply is low (<80 PSI) or

nonexistent. See “Compressed air pressure indicator” on

page 5-35.

 The user-supplied vacuum pressure is low (< 23 in Hg Vac) or

nonexistent. See “

page 5-35.

 The coupling fluid temperature is outside of the warning range:

22.1°C +

 A coupling fluid leak has been detected. See “Fluid leak indicator”

1.9°C (20.2°C – 24.0°C)

on page 5-34.

 The EMO (Emergency Motion Off) switch has been activated.

EMO indicator” on page 5-34.

See “

 The waste water bottle is full (on older Echo systems only).

See “Waste bottle indicator” on page 5-35.

Vacuum supply pressure indicator” on

3-10 Echo Liquid Handler User Manual

3.2.6 LCD screen

Emergency Off switch

There is a four-row LCD screen on the front of the Echo liquid handler and a duplicate on the rear panel. They display messages that report what the Echo liquid handler is doing or what operator action is required. Types of messages displayed include the following:

 Operating states

 Procedure progress

 Status

 Requests for operator assistance

3.2.7 Emergency Off switch

The Emergency Off switch, also known as the EMO (Emergency “Motion” Off) switch can be used in an emergency to shut down all mechanical activity in the Echo liquid handler. There are two Emergency off switches, one on the front panel and one on the rear panel of the Echo liquid handler.

Figure 3.11 Location of Emergency Off switches

 To use the Emergency Off switch in an emergency, push it in.

 To reset the Emergency Off switch, turn the knob clockwise until it

pops out (see arrows etched in the knob).

System Description 3-11

3.2.8 Anti-static bars

Lower anti-static bars

Upper anti-static bar

Right Barcode Scanner

Left Barcode Scanner

Front Barcode Scanner (not shown)

The upper and lower anti-static bars (also called ionizer bars) remove any electrostatic charge from the source and destination microplates. Electrostatic charge can affect the placement of the transferred drop during liquid transfer.

Figure 3.12 Anti-static bars

Barcode scanner

The barcode scanner is optional. You can purchase one or more barcode scanners to add to your Echo liquid handler that will read and report barcoded microplates.

Figure 3.13 Barcode scanner locations

3-12 Echo Liquid Handler User Manual

The location of each scanner depends on the location of your barcode

Left, short, or west position

Front, long, or south position

labels on the microplates. You can use either the front scanner or the side scanner, but not both at the same time.

Note: Several terms are used to specify barcode locations on a microplate. Standing in front of an Echo liquid handler, the two readable barcode locations are identified as follows:

Figure 3.14 Barcode label locations

Note: Always place side barcode labels on the left side of both source and destination microplates. The Echo liquid handler uses the left barcode scanner to read the barcode on the source plate, but uses the right barcode scanner to read the barcode on the destination plate— after it is inverted.

Labcyte provides barcode scanners that support the following barcode types:

 Code 93  Code 39  Code 128  Codabar  Pharmacode  Interleaved 2 of 5  UPC/EAN

Contact Labcyte Service and Support to install or move the barcode scanners. See “Contact information” on page 7-1.

See “Barcode Locations” on page B-1 for limitations on where barcode

labels can be placed on a microplate.

System Description 3-13

3.2.9 AC power and fuse compartment

AC Power

Fuse Compartment

Power On/Off

O = Off

I = On

RJ-45 Ethernet port

The AC power and fuse compartment (as called the power input module) is located on the rear panel, to the left of the Emergency Off button. It contains the main Echo liquid handler power on/off switch, AC power outlet, and fuse compartment.

The Echo liquid handler instruments are built in two AC power configurations: 230 VAC and 100–115 VAC.

The fuse compartment holds two fuses. For information on replacing fuses, see “Replace the AC power fuse” on page 6-18.

Figure 3.15 AC power and fuse compartment

Data connection

The Echo liquid handler includes several communication connections to the right of the Emergency off button. The RJ-45 Ethernet port is the only data connection required for normal Echo liquid handler operation. The Ethernet cable connects the Echo liquid handler directly to the client PC or to a network. This connection allows you to monitor and control the Echo liquid handler. If the Echo liquid handler is connected directly to a client PC, the Ethernet cable must be a “cross-over” cable.

Other data connection ports are used by Labcyte field engineers.

Figure 3.16 External connections: power and data.

3-14 Echo Liquid Handler User Manual

3.2.10 Coupling fluid bottle

Coupling Fluid Sensor

Coupling Fluid Bottle

The Echo system comes with a 1000 mL coupling fluid bottle. See Figure 3.17.

Figure 3.17 Coupling fluid bottle.

Note: Older Echo systems are also equipped with a 500 mL waste bottle.

Next to the coupling fluid bottle is a sensor that is connected by cable to the Echo liquid handler. The sensor monitors fluid level. When the fluid falls below a specific level, the sensor sends a low fluid volume alert to the status panel. See Figure 3.18.

Figure 3.18 Coupling fluid bottle and sensor connections.

System Description 3-15

3.2.11 Fluid chiller

Another essential component of the Echo liquid handler system is the fluid chiller. This component draws water from the coupling fluid bottle, adjusts the fluid temperature, and circulates the fluid through the acoustic transducer.

Coupling fluid temperature control

Correct Echo liquid handler coupling fluid temperature is 22.0°C. This temperature can be monitored on the DIAGNOSTICS tab of the Echo liquid handler software application. See “Coupling fluid temperature and

indicator” on page 5-36.

Set the fluid chiller to 22.0°C. See Appendix C for chiller instructions.

Caution: Incorrect fluid temperature can reduce the precision and accuracy of the fluid that is transferred, and potentially damage the acoustic transducer. Ensure that the chiller is correctly connected to the Echo instrument and is running at recommended temperature.

3.2.12 Surge tank

A 10 L surge tank is standard with the Echo liquid handler system. When vacuum demand is at its highest, the surge tank ensures a continuous source of vacuum. It prevents system errors resulting from low vacuum in laboratories where house vacuum barely meets the minimum vacuum requirement or is inconsistent due to other laboratory demands. Surge tanks are available for purchase for older Echo units.

Figure 3.19 Surge Tank

3-16 Echo Liquid Handler User Manual

3.2.13 Fluidics panel

To chiller

From chiller

Compressed

Vacuum line

air line

The fluidics panel contains tubing and hose connections to the following components:

Figure 3.20 Fluidics panel

 Chiller tubing. The chiller tubing provides the coupling fluid

circulation path between the Echo liquid handler and the chiller. The

chiller maintains the fluid temperature at 22°C.

Tubing is sufficiently long to allow the chiller to be placed on the floor

below the Echo liquid handler (for reduced vibration and good

ventilation).

 Compressed air line. The compressed air line connects an external

air supply to the Echo liquid handler. Two Echo liquid handler

mechanisms use compressed air: process door and ionizer bar.

Air pressure must be between 80 to 150 psi.

 Vacuum line. The vacuum line shown in Figure 3.20 connects the

Echo liquid handler to the surge tank, and then to the vacuum supply.

The vacuum lines inside the Echo liquid handler pull coupler fluid back

from the acoustic transducer and return the fluid to the coupler fluid

bottle. Another vacuum line removes coupler fluid from the bottom

of the source plate just before it is unloaded. Vacuum flow depends

on the vacuum source (house or pump). See vacuum specifications on

page 3-18

System Description 3-17

3.3 Specifications

Each Echo liquid handler conforms to the following specifications.

3.3.1 Physical

Height: 92.5 cm (36.4 in). Width: 53.9 cm (21.2 in). Depth: 68.3 cm (26.9 in).

Operating space envelope: Maintain a minimum operating space of

59 cm (23.2 in) width, 99 cm (39 in) depth, and 118 cm (46.5 in) height. Position the Echo liquid handler in the operating space envelope to allow

2.5 cm (1 in) side clearance (add 17.8 cm (7 in) if the coupling fluid bottle is placed on the side of the instrument), 30.5 cm (12 in) rear clearance,

25.4 cm (10 in) top clearance, and sufficient front clearance to load and unload microplates by hand or by robot. See the illustration on page 2-2.

Weight: 128 kg (283 lbs).

3.3.2 Mechanical

Air pressure: Clean, dry air at 552 kPa (80 PSI) minimum, 1034 kPa (150 PSI) maximum

Air connection: Male coupling insert (LC Series, ¼ NPT male, straight-thru) and coupling body (LC series, in-line ferruleless fitting) for 6.4 mm (¼ in) ID tubing (included in Echo liquid handler accessory kit)

Air consumption: <28.3 liter/min @ 1034 kPa (<1 cf/m @ 150 PSI)

Vacuum source:

• House vacuum: mbar, 22 in Hg Vac) Vacuum system capacity steady state: 15 L/min (0.9 m3/h, 0.53 cf/m) Drying surge:

•

Vacuum pump:

Minimum vacuum at Echo connection: 100 Torr (133 mbar, 26 in Hg Vac) Pump speed:with 10 L surge tank: 40 L/min (2.4 m3/h, 1.4 cf/m)

without surge tank (not recommended): 80 L/min (4.8 m

Note: The 10 L surge tank comes standard with the Echo liquid handler to ensure consistent vacuum.

Vacuum connection: Male coupling insert (HFC Series, 3/8 NPT male, straight-thru) and coupling body (HFC12 series, in-line hose barb) for 9.5 mm (3/8 in) ID tubing (included in Echo liquid handler accessory kit)

Minimum vacuum at Echo connection: 200 Torr (266

80 L/min for 10 sec, (4.8 m3/h, 2.8 cf/m), 33% max duty cycle.

/h, 2.8 cf/m)

Chiller performance: 22.1°C ± 0.9°C (71.8°F ± 1.6°F)

3-18 Echo Liquid Handler User Manual

3.3.3 Environmental

Temperature, operating: 21°C ± 5°C (70°F± 9°F)

Temperature, storage: 5°C–45°C (41°F–113°F) Humidity, operating: 10–80%, non-condensing

Mounting surface: Sturdy, stable, capable of handling 128 kg (283 lbs)

weight

Pollution degree: 2

Installation category: II

3.3.4 Electrical

There are two electrical configurations of the Echo liquid handler:

Config. Voltage Frequency Power Fuse

A 100– 120 VAC 50/60 Hz 10 A 250 VAC, 10 A, Fast Blow B 200–240 VAC 50/60 Hz 5 A 250 VAC, 5 A, Fast Blow

3.3.5 Client PC

Operating system: Microsoft® Windows XP SP3 or Windows 7 using 32-or 64-bit OS; Windows Net Framework 2.0

CPU: Intel® Core™ 2 Duo or later

Memory: 1GB or greater

Hard drive storage: 1GB

Video: 1280 x 1024 resolution

API: ActiveX

Programming languages: C, C++, C#, Visual Basic, Visual J#, or

Windows Net Framework compatible

3.3.6 Communications

Network connection: 10/100/1000 BaseT Network protocol: TCP/IP

User account: Labcyte (The client PC must have a Labcyte account, with

administrator privileges, that is accessible by LabcyteService and Support to service the Echo liquid handler system.)

System Description 3-19

3.3.7 Fluid transfer

Volumetric Accuracy: <10% error (single source plate-wide average from target volume)

Volumetric Precision: <8% CV

DMSO/water concentration: <8% accuracy error; <5% precision CV

(Echo 550 and 555 models only)

Fluid temperature: 21°C ± 5°C (70°F± 9°F) Compound should be in liquid form prior to transfer.

Throughput: Transfer time for fluids in different plates and on different Echo systems.

Table 3.2 Fluid throughput comparison

Plate type

1 384PP_DMSO2 2.5 1.4 2.5 2.5

2 384PP_AQ_BP2 2.5 1.8 2.9 --

3 384PP_AQ_CP 2.5 2.7 3.7 --

4 384PP_AQ_GP2 2.5 1.8 2.9 --

5 384PP_AQ_SP2 2.5 1.7 2.7 --

6 384LDV_DMSO 2.5 1.5 2.6 2.6

7 384LDV_AQ_B 2.5 1.5 2.6 --

8 1536LDV_DMSO 2.5 2.6 4.5 4.4

9 1536LDV_DMSO_HT 2.5 2.6 -- --

10 1536HB_DMSO 2.5 3.1 5.0 5.0

1 384PP_DMSO2 100 1.9 3.8 7.7

2 384PP_AQ_BP2 100 3.1 4.2 --

3 384PP_AQ_CP 100 6.0 7.2 --

4 384PP_AQ_GP2 100 3.1 4.2 --

5 384PP_AQ_SP2 100 2.2 3.3 --

6 384LDV_DMSO 100 2.0 3.9 7.7

7 384LDV_AQ_B 100 2.8 3.9 --

8 1536LDV_DMSO 100 5.8 8.2 12.7

9 1536LDV_DMSO_HT 100 4.7 -- --

10 1536HB_DMSO 100 6.2 10.2 25.5

These plate type names are used with instruments that have shipped with Echo Server 2.4. Instruments that shipped with earlier versions of software may use a different naming terminology and may have different transfer specifications. Contact your Labcyte account representative with any questions for your specific instrument.

Plate calibration 384PP_AQ_CP works with a much wider range of fluids. There are use cases (particularly in protein crystallography applications) where the volumetric accuracy may deviate from this specification. Please contact your Labcyte account representative with any questions.

Transfer volume (nL)

Echo 555

(min)

Echo 550

(min)

Echo 520

(min)

3-20 Echo Liquid Handler User Manual

Additional notes on Table 3.2

1. At low volumes, transfer times for the Echo 520 and 550 systems are similar; however, as the volume increases, the Echo 550 system shows a faster transfer time.

2. Transfer time is dependent on the number of transfers per plate; therefore, for a full plate, transfer times for a 384-well plate will always be faster than for 1536well plate.

3. At higher volumes, the difference in transfer time between the Echo systems changes significantly.

Supported fluids: Echo 550 and Echo 555 support a wide range of aqueous and DMSO-based solutions. Echo 520 system supports DMSObased solutions only.

Minimum Transferred Volume: 2.5 nL

Maximum Transferred Volume:

DMSO

 384-Well PP (polypropylene): 10 μL  384-Well LDV (low dead volume): 0.5 μL  1536-Well LDV (low dead volume): 2 μL  1536-Well LDV HT (low dead volume, high throughput): 2 μL  1536-Well HB (high-base): 2 μL

Aqueous

 384-Well PP AQ BP2 (aqueous w/buffer, protein, and up to

25% glycerol): 10 μL

 384-Well PP AQ CP (crystallography fluids): 10 μL  384-Well PP AQ GP2 (aqueous w/ glycerol at 25 to 50%): 10 μL  384-Well PP AQ SP (aqueous w/surfactant and protein): 10 μL  384-Well LDV AQ B (aqueous w/buffer only): 0.5 μL  384-Well LDV AQ P (aqueous w/protein only): 0.5 μL

Transfer Resolution: 2.5 nL

System Description 3-21

3.3.8 Supported labware

Source microplates: The following Echo qualified source microplates are available:

Table 3.3 DMSO Source Plate Types

Working vol.

Plate Calibration Vol. range (μL)

384PP 384PP_DMSO2 15-65 50

384LDV 384LDV_DMSO 2.5-12 9.5

1536HB 1536HB_DMSO 2- 6 4

(μL)

1536LDV 1536LDV_DMSO

1536LDV_DMSO_HT

HT=High Throughput. This calibration is available on Echo 555 models only. It uses the same 1536LDV plate, but with a faster transfer rate. For transfer of higher volume or where precise droplet placement is not required, see “1536-plate to 1536-plate liquid transfer” on page 1-17.

1.0 - 5.5 4.5

Table 3.4 Aqueous Source Plate Types

Plate Calibration Vol. range (μL) Working vol. (μL)

384PP 384PP_AQ_BP2

384LDV

(buffer w/protein) 15 - 65

384PP_AQ_SP2

(buffer w/surfactant &

protein)

384PP_AQ_CP

(dynamically adjusts for

surface tension/viscosity)

384PP_AQ_GP2

(aqueous w/glycerol,

from 25 to 50% volume)

384LDV_AQ_B

(buffer)

15 - 65

25-50 25

15 - 65

3 - 12 9

Volume range 15-65 is the working range but performance for sparse matrix crystallography sets will be more optimal in the 25-50 ul range.

Plate 384LDV aqueous plate precision and accuracy are only guaranteed for a maximum depletion of 500 nL from a single well without resurveying. The software will present a warning message when the depletion exceeds 500 nL, but will allow you to continue. Labcyte recommends that you test the performance of any single well depletion greater than 500 nL without a re-survey.

3-22 Echo Liquid Handler User Manual

384LDV_AQ_BP2

(buffer w/protein)

6 -14 8

Destination microplates: All 96-, 384- and 1536-well plates that meet ANSI/SBS

1-2004 standards for plate dimensions and are less than 16 mm

in height.

Short and medium flange height microplates (SBS-3, 4.1 and 4.2) are supported. Tall flange height microplates (SBS-3, 4.3) are not supported. Supported microplate height (SBS-2, 4.2.1.1) is 8.0–16 mm (0.31– 0.63 in).

Barcode types:

• Code 93

• Code 39

• Code 128

• Codabar

• Pharmacode

• Interleaved 2 of 5

•UPC/EAN

3.3.9 Coupling fluid

Fluid: Distilled water + algaecide.

Approved algaecide: Labcyte catalog number ECHO-AL04 (4 mL bottle

with dropper). Contact Labcyte to reorder.

Water/algaecide ratio: 50 μL (one drop) algaecide per 1000 mL distilled water.

ANSI=American National Standards Institute; SBS=Society for Biomolecular Sciences.

For information on plate standards, go to

http://www.slas.org/education/microplate.cfm

System Description 3-23

3-24 Echo Liquid Handler User Manual

C HAPTER

CHAPTER 0MANUAL OPERATION

This chapter describes the manual operation of the Echo® Liquid Handler, which involves a human operator performing the following procedures:

 Startup/Shutdown

 Main screen

 Create liquid transfer protocol

 Start liquid transfer run

This chapter takes the reader through the basic steps necessary to transfer sample from one plate to another with the Echo liquid handler. It is intended to present the entire range of activity in a start-to-finish flow. It is not intended to provide all of the information necessary to understand each individual step.

Refer to the chapter, “Echo Liquid Handler Software” on page 5-1, for a detailed description of each software function.

Manual Operation 4-1

4.1 Startup/Shutdown

At this point, the hardware and client PC software installations should be complete. All fluid, air, and electrical connections should be complete and functioning. Your Labcyte field engineer performs a set of function tests to ensure that the system is operating correctly.

4.1.1 Turn on Echo power

Turn on the Echo liquid handler by pressing the AC power switch located on the back panel.

The following message may appear while the Echo liquid handler initializes:

Figure 4.1 Connection failure message

Possible causes for connection failure include:

 The Echo liquid handler has not completed its initialization.

 Check the gripper stage homing sequences, particularly the theta

rotation of the destination gripper stage.

 Check the door opening sequence.

 The Ethernet cable is not connected between the client PC and the

Echo liquid handler.

 The Echo software is not configured to connect to the instrument (IP

address or hostname is missing or incorrect).

Check for the cause, attempt to resolve the problem, and try to launch the software again. If the message persists, call Labcyte Service and Support.

4-2 Echo Liquid Handler User Manual

Echo initialization

The Echo liquid handler performs a system initialization that includes the following actions:

 All three stages move to their home position:

 Source stage moves front to rear on its Y axis.  Destination stage moves left to right on its X axis, front to rear on

its Y axis, up and down on its Z axis, and partially rotates on its Theta axis.

 Acoustic transducer stage moves left to right on its X axis, front to

rear on its Y axis, and up and down on its Z axis.

Note: The LCD screen reports each step of the homing sequence as it occurs. Homing synchronizes the mechanical stages with the controlling software.

 Process door opens.  Destination stage extends, then retracts.  Process door closes.  Upper anti-static bar moves to its uppermost position, then both anti-

static bars turn off.

 Plate dryer nozzle moves to the down position.  Coupler fluid nozzle moves to the down position.  Power lamp turns on (green).  Warning lamp turns off (yellow to clear).  Fault lamp turns off (red to clear).  Coupler fluid pump turns on.  Power to the RF amplifier turns on.

Echo liquid handler warm-up

The Echo liquid handler requires a warm-up period after the power is turned on. The RF amplifier and the acoustic transducer must reach a stable operating temperature. If the instrument has been off for an extended period of time, a 60-minute warm-up period is recommended to allow all components to reach proper operating temperatures.

Notes: Echo liquid handler warm-up time depends on the starting temperature of its internal components. Always allow sufficient time for the Echo liquid handler to reach thermal stability. Fluid survey or transfer precision and accuracy will be affected by using the Echo liquid handler before it is warmed up.

Manual Operation 4-3

4.1.2 Launch Echo software

The Echo software can be launched by either of the following methods:

 Open the Labcyte Echo folder on the desktop and double-click the

icon for your Echo liquid handler.

Figure 4.2 Starting the Echo liquid handler from the Windows Desktop

 Go to START>PROGRAMS>LABCYTE>Echo 55X>Echo Liquid Handler Software.

Figure 4.3 Starting the Echo liquid handler from the Start menu

The Echo software connects to the Echo liquid handler and displays the following welcome screen.

Figure 4.4 Echo liquid handler welcome screen

4-4 Echo Liquid Handler User Manual

4.1.3 Turn off Echo power

You may need to power off the Echo liquid handler for any of the following reasons:

 Extended periods of non-use

 Preventive maintenance

 Service or repair

 Power interruptions, such as scheduled facility power interruptions

 Moving it to a new location.

When you need to turn the power off, use the following procedure:

1. Shutdown the controller inside the Echo liquid handler. From the ADVANCED window, click the SHUTDOWN button.

Figure 4.5 System reset and shutdown controls

2. Leave the vacuum supply line connected and the vacuum source turned on.

3. Switch off the AC power switch on the Echo liquid handler.

4. Wait three minutes to allow the coupling fluid to completely drain out of

the catch basin.

5. Leave the chiller on and the air pressure and vacuum lines connected.

Note: If the Echo liquid handler is going to be shut down for longer

than a week, start up and run the pump once a week to keep the acoustic transducer wet. See “Extended Non-Use and Storage” on

page 6-19.

Manual Operation 4-5

Emergency Off switch

The Emergency Off switch, also known as EMO (Emergency Motion Off) switch, can be used in an emergency to shut down mechanical Echo liquid handler activity. There are two Emergency Off switches, one on the front panel and one on the rear panel of the Echo liquid handler.

To use the Emergency Off switch in an emergency, push it in.

When to use the Emergency Off switch

 Any time the safety of the operator is in question.

 Any time strange, loud, or potentially damaging noises are coming

out of the Echo liquid handler.

 When the instrument door will not open and the source or

destination plate gripper stages collides with the door.

 If water is leaking from the Echo liquid handler.

 In any other situation that suggests immediate shutdown of the Echo

liquid handler is appropriate.

Using the Emergency Off switch

The Emergency Off switch performs the following actions:

 All motors are disabled.

 The coupling fluid pump is turned off.

 All pneumatic actuators are sent to their default positions.

 The anti-static bars are turned off.

 All RF devices (RF power supply, RF amplifier, RF switch, and so on)

are turned off.

 The HV grid is turned off.

 The internal controller aborts all processes in progress, returns an

error to the system invoking the aborted process, designates the three stages as not calibrated, and goes into a halted state.

Activating the Emergency Off switch does not turn off the following:

 Internal Echo liquid handler controller

 AC power inside the Echo liquid handler power distribution panel

 Internal 24 VDC power supply (power output, however, is

interrupted)

 External communications (Ethernet, USB)

 Coupling fluid chiller

4-6 Echo Liquid Handler User Manual

Recovering from an EMO shutdown

If the Emergency Off switch has been used to shutdown the Echo liquid handler, use the following procedure to start up again:

1. Determine and resolve the original problem.

2. Turn the Echo liquid handler back on by twisting the Emergency Off switch clockwise. It will pop out to its normal operating position.

When the Emergency Off switch is returned to its normal position, power is again supplied to most internal systems and most internal mechanisms are enabled. The Echo liquid handler stage motors, however, are not enabled yet; the operator must explicitly command a stage homing procedure (see “Motion calibration” on page 5-45

Note: If you cannot determine the original problem or you cannot correct the problem, you must arrange for a Labcyte field engineer to restore your Echo liquid handler to normal use.

Manual Operation 4-7

Tabs for each function

Protocol list In user interactive area

Server and client status window

Display options for server and client status window

Function keys

4.2 Main screen

The Echo software opens in the Main screen. The Main screen shows tabs for each major function and user interactive area for selecting from a list of protocols or clicking options or function keys. The first tab shows the PROTOCOLS window.

Figure 4.6 Below the user interactive area is the status window. This window displays every action that the Echo hardware or software performs. The follow page describes the elements of the Main screen. First window in

the Echo software

4-8 Echo Liquid Handler User Manual

 Tabs: Each tab opens a window for a function or related group of

functions.

 Protocols: In the Echo software, a protocol is a set of user-selected

parameters (plate/fluid type, plate mapping, transfer volume, and repetition) that define a single liquid transfer procedure. The PROTOCOLS window lists the protocols available to run. From this window you can create, modify, run, or delete a protocol.

 Labware: The LABWARE window shows a list of pre-defined types

of microplates. You can select these microplates during the creation of a protocol. From this window you can also create additional destination microplates.

 Diagnostics: The DIAGNOSTICS window displays the status of the

Echo liquid handler. From this window you can also manipulate some of the individual motor controls and perform some maintenance tasks.

 Calibration: The CALIBRATION window provides calibration

procedures for microplate registration, power modulation, transducer focus, and barcode alignment.

 Advanced: The ADVANCED window provides additional liquid

transfer options and system functions, such as shutting down or restarting the instrument.

 Function Keys: Function keys perform tasks specific to the window

in which they are displayed, such as creating, editing, or removing a protocol.

 Status window: Two status windows are available: Client and

Server. The Client status window shows the applications that are executed during a protocol run. The Server status window shows the individual tasks that are performed within an application.

The status windows also display specific types of errors that the system encounters during a protocol run.

 About and Help menu (not shown): Right-clicking on a tab window

background, will display a menu that provides two choices: About and Help.

About: Select ABOUT to display the software version and build numbers.

Help: Select H

Each function window is described in more detail in Chapter 5, "

ELP or press the F1 key to open the online Help file.

Echo

Liquid Handler Software"

Manual Operation 4-9

4.3 Create liquid transfer protocol

The liquid transfer protocol specifies how much fluid will be transferred from the source microplate to the destination microplate, and in what well order

—a plate map that the Echo instrument uses to perform the transfer.

For example, a simple transfer protocol would transfer compound from all source wells to the matching destination wells, replicating the contents of the source plate. More complex protocols would transfer compound from different source plates to one or more destination microplates. The Echo liquid handler software provides a plate map to specify the wells that will be used in the liquid transfer. For a detailed discussion of different types of liquid transfer protocols, see “

supported?” on page 1-12.

Note: To create transfer protocols for specific applications or specialized plate configuration, such as dose-response or protein crystallization, you can purchase optional software applications or the Echo Software Development Kit to create your own application.

See “Echo applications” on page 1-19.

Sample protocol parameters

What types of liquid transfer are

The example in the following procedure uses these parameters:

 Source microplate:384LDV_AQ_P

(384-well, COC, low dead volume microplate for aqueous protein solution)

 Destination microplate: An empty 384LDV microplate

 Plate map: The twenty wells defined as A1 through E4

 Copies: 5

 Volume: 10 nL transfer into each well

 Source well contents: Sample fluid consisting of 1 mg/mL protein

diluted in aqueous buffer.

4-10 Echo Liquid Handler User Manual

4.3.1 Add new protocol

1. From the PROTOCOLS window, click the ADD button. The New Protocol wizard will begin.

2. Enter the protocol name (for example, 384LDV Aqueous P 10 nL) and select the source and destination microplate types from the drop-down lists.

Figure 4.7 Entering protocol name

3. Click the NEXT button. The second window will open.

Figure 4.8 Add new protocol wizard, page 2

4. Enter the number of copies (for example, 5). Each copy represents a single plate transfer.

5. Enter the liquid transfer volume (for example, 10 nL). If you enter the transfer volume directly instead of using the increment/decrement buttons, remember that the value must be an even multiple of the minimum transfer volume (2.5 nL). If you enter any other value, you will see an error message.

Manual Operation 4-11

6. Click the V

IEW/DEFINE button to display the DEFINE WELLS window.

Figure 4.9 Defining transfer wells

7. Specify the source wells from which to transfer fluid. To define the region A1 through E4, click cell A1 and drag the mouse pointer to cell E4.

8. Click the OK button to return to the previous window.

9. Click the FINISH button to store your new protocol definition.

For more information about creating a 384-well protocol, see “Create a

new 384:384 protocol definition” on page 5-5.

4-12 Echo Liquid Handler User Manual

4.4 Run the liquid transfer protocol

Before running the liquid transfer protocol, you will need to prepare the compound to be transferred and set up the microplates.

4.4.1 Prepare the microplates

Prepare the microplates as follows:

1. Prepare the source microplates. Consider the following factors:

 Fluid height for a given fluid volume can vary, depending on the

surface interaction of the fluid with the well walls.

 Fluids that cling to microplate plastics will form steep menisci,

pulling fluid away from the center of the wells, causing lower fluid height.

 Fluids that interact less with the well walls will form shallow

menisci, leaving more fluid in the center and greater fluid height.

 Fluids that contain higher concentrations of protein, or higher

concentrations of detergents, may also form steeper menisci. Since there is a wide variety of fluid compositions, you may find that the Echo liquid handler reads a different volume

 Bubbles in the fluid prevent the formation of a fluid meniscus. For

optimal results, remove any bubbles in the wells and check for welldefined menisci.

Note: Spinning down the microplate can remove bubbles and even out variations in menisci. Use 1000 x g for DMSO samples; 3000 x g for aqueous samples.

 If the source microplates were made earlier and stored refrigerated or

frozen, allow them to equilibrate or thaw to room temperature.

 Aqueous solutions evaporate quickly at low volumes. For longer runs,

fill wells to the maximum volume specified for the source plate type used. Volume ranges may differ for different aqueous solutions as well as aqueous vs. DMSO-based solutions.

 DMSO at 100% absorbs water rapidly from the atmosphere and can

increase the final volume of the compound in the source microplate, which might exceed the maximum volume allowed by the Echo liquid handler. Fill plates below the maximum volume to account for hydration. This may be dependent on your laboratory conditions and frequency of source plate use.

Manual Operation 4-13

2. Place the appropriate plate insert onto the plate gripper (if it is not

already in position). See “

3. Collect the microplates that will serve as destination plates to receive the compound. A destination microplate definition must exist or be created. For example, knowing the plate height of the destination microplate allows the Echo liquid handler to lower the destination microplate into very close proximity to the source microplate, improving droplet ejection accuracy and reliability.

Source plate insert” on page 3-6.

4.4.2 Start liquid transfer run

You are ready to run the liquid transfer protocol:

1. From the PROTOCOLS window, select the protocol to be run. If you move the mouse pointer over a protocol name, a tool tip displays some of the protocol settings.

Figure 4.10 Protocols window

2. Click the RUN button. You will be instructed to click the NEXT button. The process door will open and the source plate gripper stage will extend outside of the Echo liquid handler.

4-14 Echo Liquid Handler User Manual

Figure 4.11 Starting transfer protocol

3. Load the source microplate with well A1 in the inner-left corner. Click the N

EXT button.

Figure 4.12 Loading source plate

The source plate gripper stage retracts into the Echo liquid handler and the destination plate gripper stage is extended outside of the Echo liquid handler.

4. Load the destination microplate with well A1 in the inner-left corner. Click the N

EXT button.

Figure 4.13 Loading destination plate

Manual Operation 4-15

While the destination plate gripper stage inverts the microplate, the source microplate is surveyed. The Echo liquid handler determines the fluid composition and fill height, and then calculates the fluid volume in each well. For aqueous solutions, the fluid volume is an approximation and may vary depending on protein and surfactant concentrations, among other factors.

After the source plate survey is done and the destination microplate is positioned correctly above the source microplate, the liquid transfer process begins.

Using the data contained in your destination microplate definition, the protocol definition, and the source microplate survey just performed, droplets are ejected from each specified source microplate well into the corresponding destination microplate well. A progress bar keeps you informed about the liquid transfer that is in progress.

If you need to abort the liquid transfer process, click the C

ANCEL button.

5. When the liquid transfer is complete, click the NEXT button.

Figure 4.14 Transfer in progress

The destination plate gripper stage extends outside of the Echo liquid handler and reverts the plate to its upright position.

6. Remove the destination microplate and click the NEXT button.

4-16 Echo Liquid Handler User Manual

Figure 4.15 Removing destination plate

The destination plate gripper stage retracts into the Echo liquid handler and the source plate gripper stage extends outside of the Echo liquid handler.

7. Remove the source microplate and click the D

ONE button.

Figure 4.16 Removing source plate

The source plate gripper stage retracts into the Echo liquid handler.

The liquid transfer protocol is done.

Manual Operation 4-17

4-18 Echo Liquid Handler User Manual

Labcyte Echo 520, Echo 550, Echo 555 User Manual

Specifications and Main Features

Frequently Asked Questions

User Manual

Intended audience

Preface vii

Safety warnings and precautions

Safety notation marks

Electrical safety warnings

Additional safety warnings

Safety cautions

Emergency Off switch (EMO)

Laser safety warnings

Product labels

Intellectual property label

ETL label

Laser safety labels

Electrical hazard

General warning label

Water system maintenance label

WEEE label

Emergency Off label

Laser safety labels

Product label

Pinch point label

High-voltage label

Using this manual

Who should read this manual?

What is in this manual?

Conventions used in this manual

Introduction 1-1

1.1 Acoustic Droplet Ejection

1.1.1 How does the Echo system work?

1.1.2 Will the fluid splash or fall out?

1.2 What are the Echo liquid handler features?

1.2.1 “Touchless” fluid transfers

1.2.2 Reliability

1.2.3 Ease of use

1.2.4 System integration friendly

1.2.5 Low maintenance requirements

1.2.6 Cost savings

1.2.7 High accuracy, precision, and speed

1.2.8 Value-added quality control

1.3 What types of liquid transfer are supported?

1.3.1 384-plate to 384-plate liquid transfer

1.3.2 384-plate to 96-plate liquid transfer

1.3.3 384-plate to 1536-plate liquid transfer

1.3.4 1536-plate to 384-plate liquid transfer

1.3.5 1536-plate to 1536-plate liquid transfer

1.4 What system configurations are supported?

1.4.1 Manual operation

1.4.2 Echo applications

1.4.3 Modular workstation operation

1.4.4 Small cell operation

1.4.5 Fully automated operation

1.4.6 Multi-client considerations

Installation and Relocation 2-1

2.1 Pre-installation requirements

2.1.1 Site requirements

2.1.2 Additional components

2.2 Installation overview

2.3 Software installation

2.3.1 Install the software

2.3.2 Uninstall the Echo software

2.3.3 Upgrade the Echo software

2.4 Instrument Relocation

2.4.1 Prepare the Echo liquid handler for a move

System Description 3-1

3.1 System overview

3.1.1 Front panel

3.1.2 Back panel

3.2 Component description

3.2.1 Process door

3.2.2 Source plate gripper stage

3.2.3 Source plate insert

3.2.4 Destination plate gripper stage

3.2.5 Status indicator lights

Power light

Warning light

Fault light