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A-6

Echo Liquid Handler User Manual

A.4.4 Examples of acoustic droplet ejection

Figure A.3 shows a 5 nL droplet being ejected from a volume of water. The 
acoustic energy pulse creates a mound of fluid at the surface, which is 
pushed up. A droplet forms on a neck and eventually pinches off. Its 
momentum carries it up to a receiving surface while the fluid mound 
recedes into the larger fluid volume.

Figure A.3  A droplet ejection

Summary of Contents for Echo 520

Page 1: ...For Echo models 520 550 555 ...

Page 2: ...ates 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 ...

Page 3: ... 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 maintenan...

Page 4: ...tware 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...

Page 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 Calibrati...

Page 6: ...rage 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 th...

Page 7: ... 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 Echo liquid handler operators must read this manual before ...

Page 8: ...tended 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 cond...

Page 9: ...he 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 ...

Page 10: ...tempt 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...

Page 11: ...ithout 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 ...

Page 12: ...tion 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 in...

Page 13: ...operty 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 ...

Page 14: ...instrument back cover This warning indicates that the Echo liquid handler optionally uses laser based barcode scanners The Echo liquid handler is considered a Class 1 laser light hazard under normal operating conditions with instrument covers on Electrical hazard The electrical hazard label warns of possible injury from electrical cables connections and circuits inside the Echo liquid handler Gene...

Page 15: ... 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 Emergency Off label The Emergency Off label identifies the EMO Emergency Motion 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 f...

Page 16: ...th 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 The pinch point label warns of possible injury if you do not keep your hands or fi...

Page 17: ...Preface xvii High voltage label The high voltage label warns of possible high voltage injury from electrical cables connections and circuits inside the Echo liquid handler ...

Page 18: ... 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 ...

Page 19: ...atting to help indicate emphasis or keystrokes The text conventions are as follows Convention Example Meaning Small caps bold Press the NEXT button Indicates an on screen button label menu title or menu item Courier client txt client old2 txt Indicates a display output printed output keyboard input or file names Blue underlined Software installation Signifies a hyperlink to a topic Quotation marks...

Page 20: ...xx Echo Liquid Handler User Manual ...

Page 21: ... 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 ...

Page 22: ... 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 r...

Page 23: ...e 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 ...

Page 24: ...e 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...

Page 25: ...plets 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 ...

Page 26: ... 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 follo...

Page 27: ...ls 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 trans...

Page 28: ...nation 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 pro...

Page 29: ...isms 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...

Page 30: ...s 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 compa...

Page 31: ...olutions 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 par...

Page 32: ... Table 1 1 Liquid transfer types Source Src Microplate Destination Dest Microplate Transfer map type full or partial 384 well 96 well destacking1 quadrant 4 regions from 1 src plate to 4 dest plates 1 Destacking transferring liquid from a single source src plate to several destination dest plates destacking interleaved 2x2 or every other dest well 384 well 384 well standard 1 src plate to 1 dest p...

Page 33: ...ombination 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 ...

Page 34: ...pplications 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 plate...

Page 35: ...ate 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 interfac...

Page 36: ...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 ...

Page 37: ... 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 ...

Page 38: ...eration 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 1 4 1 Manual operation Manual operation leaves control of every step to the operator Defining new fluid transfer protocols or new destination mi...

Page 39: ...enerates 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 labwa...

Page 40: ...s 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 rob...

Page 41: ...n 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 ...

Page 42: ...ination 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 No...

Page 43: ... 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 to day work flo...

Page 44: ...arantees 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 be...

Page 45: ...ional 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 ...

Page 46: ... x 99 cm 39 in depth x 118 cm 46 5 in height Figure 2 1 Instrument layout on bench top 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 ...

Page 47: ...m 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 I...

Page 48: ...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 ...

Page 49: ...tle 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 Co...

Page 50: ...nto 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 serve...

Page 51: ...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...

Page 52: ...he 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 ...

Page 53: ...e 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 transfe...

Page 54: ... 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 ...

Page 55: ... 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...

Page 56: ...he 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 Coupler fluid bottle cap blue bottle cap From vacuum source To coupler catch nozzle long straight tube From catch basin J tube From chil...

Page 57: ... 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 Fluidics panel From chiller Water In To chiller Water Out Compressed air lin...

Page 58: ...2 14 Echo Liquid Handler User Manual ...

Page 59: ... 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 flu...

Page 60: ...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 ...

Page 61: ...rmation 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 Emergency Motion Off switch shuts down all mechanical Echo liquid handler activity Th...

Page 62: ... 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...

Page 63: ... 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 assemblie...

Page 64: ...ed 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 Note Handle the plate inserts carefully They can be damaged from dropping or rough handling which can affect instrument performance Table 3 1 Labcyte plate inserts Plate insert Plate catalog Inse...

Page 65: ... 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 ...

Page 66: ...rred 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 re...

Page 67: ...s 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 pa...

Page 68: ...d 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 tur...

Page 69: ... 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 ...

Page 70: ...ect 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 Lower anti static bars Upper anti static bar Right Barcode Scanner Left Barcode Scanner Fron...

Page 71: ...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 Contac...

Page 72: ...igure 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 a...

Page 73: ...so 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 Coupling Fluid Sensor Coupling Fluid Bottle ...

Page 74: ... 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 handle...

Page 75: ...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...

Page 76: ... connection Male coupling insert LC Series NPT male straight thru and coupling body LC series in line ferruleless fitting for 6 4 mm 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 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...

Page 77: ...2 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 adminis...

Page 78: ...ype1 Transfer volume nL Echo 555 min Echo 550 min Echo 520 min 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...

Page 79: ...555 support a wide range of aqueous and DMSO based solutions Echo 520 system supports DMSO based 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 ...

Page 80: ... Types Plate Calibration Vol range μL Working vol μL 384PP 384PP_AQ_BP2 buffer w protein 15 65 50 384PP_AQ_SP2 buffer w surfactant protein 15 65 384PP_AQ_CP1 dynamically adjusts for surface tension viscosity 25 50 25 384PP_AQ_GP2 aqueous w glycerol from 25 to 50 volume 15 65 50 384LDV2 384LDV_AQ_B buffer 3 12 9 384LDV_AQ_BP2 buffer w protein 6 14 8 1 Volume range 15 65 is the working range but per...

Page 81: ...eight 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 2 ANSI American National Standards In...

Page 82: ...3 24 Echo Liquid Handler User Manual ...

Page 83: ...ol 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 p...

Page 84: ...ear 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...

Page 85: ...ition 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 turne...

Page 86: ...e 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 ...

Page 87: ...uid 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 conne...

Page 88: ...y 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 o...

Page 89: ... 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 o...

Page 90: ...on 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 Tabs for each function Protocol list In user interactive area Server and client status window Display options for ser...

Page 91: ...s 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 win...

Page 92: ...e liquid transfer For a detailed discussion of different types of liquid transfer protocols see What types of liquid transfer are 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...

Page 93: ...ng 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 m...

Page 94: ...rce 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 ...

Page 95: ...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 well defined 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 earl...

Page 96: ...quid handler to lower the destination microplate into very close proximity to the source microplate improving droplet ejection accuracy and reliability 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 ...

Page 97: ...the NEXT 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 NEXT button Figure 4 13 Loading destination plate ...

Page 98: ... 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 liqui...

Page 99: ...s 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 DONE button Figure 4 16 Removing source plate The source plate gripper stage retracts into the Echo liquid handler The liquid transfer protocol is done ...

Page 100: ...4 18 Echo Liquid Handler User Manual ...

Page 101: ...clude the following Protocols window defines new liquid transfer protocols and edit or delete existing protocols run the protocols Labware window defines new destination plates and edit or delete existing destination plate definitions Diagnostics window monitors Echo liquid handler status directly control most of the Echo liquid handler motor function and perform diagnostic procedures Calibration ...

Page 102: ...arameters From the PROTOCOLS window you can perform the following tasks ADD create a new liquid transfer protocol definition EDIT change an existing protocol definition REMOVE delete an existing protocol definition RUN execute a liquid transfer protocol Figure 5 1 The Protocols window Echo software can be used to create several types of transfer protocols Examples of protocols that can be created ...

Page 103: ...src plate to 4 dest plates destacking interleaved 2x2 or every other dest well 384 well 384 well standard 1 src plate to 1 dest plate 384 well 1536 well stacking2 quadrant 4 src plates to 4 regions on 1 dest plate stacking interleaved 2x2 or every other dest well 384 well 3456 well stacking quadrant 9 src plates to 9 regions on 1 dest plate stacking interleaved 3x3 or every third dest well 1536 we...

Page 104: ...umber of transfers and the specific wells involved Protocols are stored in the Echo database and displayed every time you view the PROTOCOLS window From the PROTOCOLS window you can click the ADD button to begin the Add New Protocol wizard After you have created the protocol you can select it in the Protocol List see Figure 5 2 to run edit or remove Figure 5 2 The protocol list Samples of protocol...

Page 105: ...EW PROTOCOL WIZARD Enter the following information Protocol Name Enter a name for the new protocol It must not already be in use Source Plate Type Select a source plate type from the first drop down list Note Only Echo qualified source microplates are listed in the Source Plate Type drop down list Contact Labcyte to add more source plates See Contact information on page 7 1 Destination Plate Type ...

Page 106: ...e plate wells that contain the compound to be transferred See Figure 5 5 For information on how to select transfer wells see Create a transfer map on page 5 14 b Click the OK button to return to the previous screen Figure 5 5 The Define Wells dialog box 4 Number of Copies Enter the number of times the protocol will be executed The valid range is 1 to 10 000 copies The operator is prompted to load ...

Page 107: ...ents of 2 5 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 If you type an invalid transfer volume you will receive an error message that displays the correct transfer range Figure 5 6 Invalid transfer volume for 384 well plate 6 Click the FINISH button to complete the new ...

Page 108: ...ew 384 1536 protocol definition Create a new 384 1536 protocol definition using the stacking features of the ADD NEW PROTOCOL WIZARD Compound from four 384 well source plates can be transferred to a single 1536 well destination plate See 384 plate to 1536 plate liquid transfer on page 1 15 for an illustration of plate mapping to more than one destination plates Note If the new liquid transfer prot...

Page 109: ...he protocol will be executed The valid range is 1 to 10 000 copies You are prompted to load and unload the source microplate and the specified number of destination microplates Transfer Volume Enter the total volume of fluid to be transferred The valid range is dependent on the source microplate see Maximum Transferred Volume on page 3 21 For example for the Labcyte 384 well PP source microplate t...

Page 110: ...ton to create a transfer map for the first source plate The Define Wells dialog box shows the wells that are available for transfer Select the wells you want to include in the transfer Repeat this step for the other source plates and click the OK button Figure 5 12 Define wells in a transfer map For an explanation on how to select wells in a transfer map see Create a transfer map on page 5 14 5 Cl...

Page 111: ...res a new destination microplate definition create it before running the ADD NEW PROTOCOL WIZARD See Manage labware specifications on page 5 24 1 Click the ADD button to start the ADD NEW PROTOCOL WIZARD 2 Protocol Name Enter a name for the new protocol It must not already be in use a Source Plate Type Select a source plate type from the first drop down list Note A 1536 well source microplate must...

Page 112: ...he number of times the protocol will be executed The valid range is 1 to 10 000 copies You are prompted to load and unload the source microplate and the specified number of destination microplates Transfer Volume Enter the transfer volume in 2 5 nL increments up to 2000 nL or up to 500 nL for aqueous transfers If you enter the transfer volume directly instead of using the increment decrement butto...

Page 113: ... Wells dialog box shows you the source wells white cells that can be transferred to the selected destination plate The gray cells represent the wells that are not available If you click the radio button for the different destination plates you will see that the transfer map changes for each destination plate Figure 5 18 Transfer maps for all four destination plates a Click the radio button for Des...

Page 114: ... maps for Dest 1 to Dest 4 6 Click the FINISH button after all four transfer maps have been created Create a transfer map A transfer map must be created for each source microplate The transfer map defines which source microplate wells contain compound that will be transferred to the destination plate When you click the VIEW DEFINE button the DEFINE WELLS dialog box opens Figure 5 19 The Define Wel...

Page 115: ...elected Click on a row label or a column label The entire row or column will be selected Click and drag the mouse point to define a rectangle of wells in one move If a group of wells is selected and another well or group of wells must also be selected hold the CONTROL key down so that the original group of wells is not cleared All wells can be selected by clicking in the upper left square above th...

Page 116: ...ouble click the protocol name in the PROTOCOL LIST One of the following edit protocol dialog boxes will open Figure 5 21 Edit 384 384 protocol dialog box Figure 5 22 Edit 384 1536 protocol dialog box Any parameter of a user defined protocol can be edited however the destination plate type cannot be changed from 384 well to 1536 well or vice versa ...

Page 117: ...l is required The protocol definition is deleted from the Echo database and from the Echo software PROTOCOL LIST 5 1 4 Run a protocol To run a liquid transfer protocol 1 From the PROTOCOLS window select the protocol to be run and click the RUN button The first of a series of screens is displayed to provide instructions to run the selected protocol Figure 5 23 Protocol selected to be run ...

Page 118: ... 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 your destination microplate with well A1 in the inner left corner Click the NEXT button While the destination plate gripper stage retracts the microplate the source microplate is surveyed The Echo liquid handler determines the fluid composition an...

Page 119: ...f the liquid transfer in progress You can abort the liquid transfer process by clicking the ABORT button 5 When the liquid transfer is complete click the NEXT button 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 The destination plate gripper stage retracts in...

Page 120: ...urce plate specifications cannot be added Edit To edit a plate specification either select the plate name and click the EDIT button or double click the plate name to open the PLATE SPECIFICATION window Editing source plate specification editing is limited to selecting a barcode location including none Destination plate specification editing is not restricted See the section Manage labware specific...

Page 121: ...h 384 well LDV and 1536 well LDV plates are manufactured by Labcyte to meet tight plate tolerances for lower volume applications Example 2 384PP_AQ_BP2 384 well polypropylene _ aqueous_buffer _ 2nd generation vol range This fluid type is available in Echo 550 555 models that ship with software version 2 4 and later Volume range is 15μL to 65 μL Example 2 384PP_AQ_CP 384PP_AQ_GP2 384 well polypropy...

Page 122: ...s destination plate can accept sample from any Echo qualified source plate Example 2 Greiner_1536HiBasePS_782096 vendor name which includes 1536 well polystyrene In this example the vendor name is used to describe the plate and there is no Dest ending Operators can create destination labware at any time and adopt whatever is the most convenient convention for their lab Laboratories using multiple ...

Page 123: ...ge 7 1 Note In Echo application software such as Echo Dose Response and Echo Cherry Pick version 1 5 or later existing source plates can be copied and renamed to allow for more specific identification for application workflows and then imported into Echo software However other plate parameters still cannot be changed except for barcode location Figure 5 25 Plate Specification for Source Plate Sour...

Page 124: ... the source plate specification looks the same but only the barcode location can be edited Figure 5 26 Plate Specification for Destination Plate Plate Name Enter a plate name to identify the new specification required Manufacturer Enter the manufacturer s name Lot Number Enter the plate lot number identification Part Number Enter the manufacturer s plate part number Rows Enter the number of rows T...

Page 125: ...s may already be specified in the SBS standards see SBS standards on page 5 27 Figure 5 28 Plate dimensions A B C and D A and B Well A1 X Y offsets Dimensions A and B locate the center of well A1 in relation to the plate s left and top outside edges A specifies the A1 well center to the left outside edge in millimeters B specifies the A1 well center to the top outside edge in millimeters Examples ...

Page 126: ...e valid range for C is 0 05 mm to 9 00 mm Refer to ANSI SBS 4 2004 for more information D Y center spacing Dimension D specifies the vertical distance between well centers in millimeters That is the distance between wells A1 B1 C1 and so on Example For a 1536LDV DMSO plate type D is 2 25 mm The valid range for D is 0 05 mm to 9 00 mm Refer to ANSI SBS 4 2004 for more information Figure 5 30 Plate ...

Page 127: ...d for source plates and is not necessary for destination plates Example For a 384PP_DMSO plate type G is 3 3 mm The valid range for G is 0 00 to 86 00 mm H Well volume Dimension H specifies the volume of the well in microliters Example For a 384PP_DMSO 50 0 μL The valid range for H is greater than 0 0 μL Note The Echo liquid handler does not currently use the destination well volume property and a...

Page 128: ...dicators Clicking the check box next to the device initiates the action for example clicking Source Plate In directs the Echo liquid handler to retract the source plate gripper stage In the SYSTEM STATUS section the user can monitor the current overall status of the Echo liquid handler through ten additional status indicators and two internal Echo temperature readouts In the MISCELLANEOUS section ...

Page 129: ...5 38 The MOTION section of the CALIBRATION window uses the HOME and REGISTER buttons See Motion calibration on page 5 45 Note The Echo liquid handler will not allow device movement that would cause damage to itself For example if the process door is closed when the source plate gripper stage is commanded to extend outside the Echo liquid handler opens the process door first Figure 5 32 Device cont...

Page 130: ... the source plate One light reports the current position in out of the source plate gripper stage the other light reports the presence of a microplate loaded onto the plate gripper The plate presence indicator has two states Clear Plate not present Green Plate present Destination plate in out and presence The destination plate gripper stage can be commanded to extend outside or to retract back ins...

Page 131: ...ndler when commanded to flip upright a Flip Dest Plate Up Error will appear The associated indicator lights report the current rotational position of the destination plate gripper stage Plate dryer up down on off The Echo liquid handler can be commanded to raise or lower the plate dryer nozzle or to turn its vacuum supply on or off Select the appropriate checkboxes If necessary the Echo liquid han...

Page 132: ...r anti static bar Coupler nozzle up down The Echo liquid handler can be commanded to raise or lower its coupler fluid nozzle also called the bubbler at any time The associated indicator lights report the position of the coupler fluid nozzle Coupler pump on off The Echo liquid handler can be commanded to turn the coupler fluid pump on or off The associated indicator lights report whether the couple...

Page 133: ... The Echo liquid handler is not powered on The Echo internal controller is not running The Echo liquid handler is still initializing after being powered on and is not yet ready for use Warning indicator The WARNING status indicator reports conditions that will not allow normal operation Gray OK The Echo liquid handler is ready for use Yellow One or more of the following is true The Echo stages are...

Page 134: ...n page 5 34 The waste bottle is full older Echo systems only See Waste bottle indicator on page 5 35 EMO indicator There are two EMO Emergency Motion Off switches one on the front and one on the rear of the Echo liquid handler Gray OK The Emergency Off switches are in their normal not activated position Red An Emergency Off switch has been activated Power has been turned off to many internal Echo ...

Page 135: ...tions or for the vacuum turned off at the source Coupling fluid indicator A sensor monitors the level of coupling fluid in its bottle Green OK Coupling fluid is above the minimum required level Yellow The coupling fluid bottle level is low 250 mL Note When the coupling fluid falls below its recommended minimum level bubbles may be introduced into the coupling fluid reducing the accuracy or precisi...

Page 136: ...the temperature of the acoustic transducer A correct and stable acoustic transducer temperature is required to maintain accurate and precise liquid transfers The coupling fluid temperature indicator reports three states Green Coupling fluid temperature is OK Yellow Coupling fluid temperature warning The coupling fluid temperature is outside of the normal range 21 2 C 23 0 C Red Coupling fluid temp...

Page 137: ...on off Coupler nozzle up down Executing the dry plate diagnostic procedure Executing the move to well diagnostic procedure Executing the perform survey diagnostic procedure Homing the stages Power calibration Acoustic transducer calibration Barcode scanner calibration The following Echo functions continue to be available during maintenance mode Creating editing or deleting fluid transfer protocol ...

Page 138: ...way and the coupling fluid pump is turned off The plate dryer nozzle makes a pass to ensure that all of the moisture is removed then displays a message when it is done Move to well The MOVE TO WELL procedure positions the acoustic transducer directly beneath a specified well 1 Select MOVE TO WELL and click the LAUNCH button The MOVE TO WELL dialog box is displayed 2 Enter the row and column addres...

Page 139: ...urce plate 1 Go to the DIAGNOSTICS window and use the Diagnostics controls to load a source plate into the Echo liquid handler 2 Select Survey from the MISCELLANEOUS drop down list 3 Click the LAUNCH button The SURVEY window will open Figure 5 35 Survey procedure 4 From the NAME drop down list select the correct source plate type ...

Page 140: ... automatically store the results of the acoustic survey in the client PC data directory C Program Files Labcyte Medman Data Also see File Save on page 5 42 Ignore Source Plate Sensor Select this checkbox to resolve problems with fitting a source plate onto the source plate gripper stage This option allows the Echo liquid handler to use a source plate without detecting its presence with the source ...

Page 141: ...in MRayl Fluid Thickness Displays well fluid thickness Post Processing Raw The measured fluid percentages are not processed Processed homogeneous The measured fluid percentages are processed using a moving average filter Processed inhomogeneous The measured fluid percentages are processed to reduce measurement noise Note Selecting processed or raw fluid percentages affects the following data The d...

Page 142: ...S section the minimum maximum average and CV data from the survey is reported These statistics are calculated from raw processed homogeneous or processed inhomogeneous survey data depending on which POST PROCESSING radio button is selected In the RESULTS section raw data collected during the survey is displayed This information is used only by Labcyte personnel during troubleshooting or system ana...

Page 143: ...at FILE SAVE FLUID COMPOSITION will save raw processed homogeneous or processed inhomogeneous survey data depending on which POST PROCESSING radio button is selected The SAVE AS dialog box is displayed Figure 5 39 Diagnostic survey file saved ...

Page 144: ...l 5 4 Calibration window Several Echo mechanisms can be user calibrated The following motion power acoustic transducer and barcode scanner related calibration procedures are found in the CALIBRATION window Figure 5 40 Calibration window ...

Page 145: ...ft right Y front rear and Z up down During the homing procedure the destination plate gripper stage extends outside of the Echo liquid handler to rotate safely Figure 5 41 Homing the stages After homing its stages the Echo controller knows the absolute position of each stage Register calibration The REGISTER button calibrates the well position for the selected source plate and updates the labware ...

Page 146: ...3 Click the PRESENT button to extend the source plate gripper stage 4 Place the source plate on the plate gripper 5 Click the RETRACT button to draw the plate into the instrument 6 Click the REGISTER button The Echo liquid handler calibrates the well position and displays the new well center values 7 Click the APPLY LABWARE button at the bottom of the dialog box then YES in the Confirmation Update...

Page 147: ...will not need to be performed by a user However it can be useful during certain repair diagnostic or recovery procedures Perform power calibration only as directed by Labcyte personnel If manual power calibration is required use the following procedure 1 On the CALIBRATION tab click the CALIBRATE button in the POWER section The CALIBRATE POWER dialog box will open Figure 5 44 Calibrate Power dialo...

Page 148: ...E button in the POWER section of the CALIBRATE POWER dialog box The Echo liquid handler performs the following procedure The most recent power calibration record stored in the Echo database is read This data will be used in the next step An RF pulse is generated Several RF measurements are taken A new amplifier feedback value is calculated and displayed as POWER AMPLIFIER FEEDBACK A new pulse ener...

Page 149: ...performed and or the SYSTEM GAIN parameter has been modified there are three actions to choose from OK The OK button applies the new AMPLIFIER FEEDBACK value and or the new System Gain value and closes the CALIBRATE POWER dialog box Change confirmation is not requested Apply The APPLY button displays a dialog box asking for confirmation of the AMPLIFIER FEEDBACK and or SYSTEM GAIN change s Answeri...

Page 150: ...ect droplet ejection requires the ability to accurately focus the acoustic energy in the well fluid Changes to the transducer focal length over time may lead to a mispositioning of the Z axis Measurement of the DMSO water ratio during a source plate survey will be affected In turn inaccurate DMSO water ratio measurement can degrade the accuracy of fluid droplet ejection Calibrating the Echo acoust...

Page 151: ...ues and do not select the OVERRIDE CALCULATED RANGE checkbox 3 Click the NEXT button The Echo process door opens the source plate gripper stage extends and the Echo liquid handler performs a focal sweep The focal sweep uses acoustic pings bounced off an internal registration plate while the transducer is being moved vertically The focal sweep process measures the transducer s focal length After th...

Page 152: ...libration page 2 new focal length calibration not required Note If your graph shows two curves the Echo software has detected older calibrations software version 2 2 and older and has performed a focus calibration check for both current and older calibrations ...

Page 153: ...ch liquid transfer protocol that is run Each setting has a default state enforced when the Echo software is launched or restarted User choices are not stored or remembered upon shutdown Survey interval Select the interval between source plate surveys by choosing a value from the PERFORM EVERY SURVEY drop down list The default setting of every 1 plate causes the source plate to be surveyed every ti...

Page 154: ...urce microplate is one where the solution to be transferred is very similar well to well If the USE PREDEFINED checkbox is selected the Assume Homogeneous Plate feature is unavailable and its checkbox is grayed out Default Assume homogeneous plate off Power Monitoring Selecting the POWER MONITORING checkbox causes the Echo liquid handler to monitor the RF power used to eject each fluid droplet Lab...

Page 155: ...etect sensor is not ignored Ignore Destination Plate Sensor Check Selecting the IGNORE DESTINATION PLATE SENSOR CHECK checkbox bypasses the destination plate present sensor This may be useful when an individual destination plate does not fit correctly or droplets are being transferred to a microplate lid or other substrate that does not fit like a microplate A run protocol procedure can execute wi...

Page 156: ...ll of the plate concentration map and enter the DMSO concentration for that well For example if the fluid contains 72 5 DMSO enter 72 5 Figure 5 50 Solvent platemap Solvent Concentration Percentage Enter a DMSO concentration in preparation for clicking the FILL button Plate Type Select the source microplate type Fill Clicking the FILL button loads the entered DMSO concentration percentage into eac...

Page 157: ...e running on the client PC clicking the HARD RESET button will reset the client PC and clicking the SHUTDOWN button will shutdown the client PC Soft Reset Clicking the SOFT RESET button causes the Echo service to be restarted on the Echo controller Hard Reset Clicking the HARD RESET button causes the Echo controller to reboot After rebooting the Echo controller any attached client computer program...

Page 158: ...n for the client log files is C Program Files Labcyte Medman Data The most recent client log file is named client_localhost_MedmanGUI txt and older client log files are named client_localhost_MedmanGUI old 0 txt client 1 txt client 2 txt and so on The most recent ten historical client log files are stored in addition to the log file for the current session Server log messages Messages in the serve...

Page 159: ...ssages are generally not of use to Echo users They provide detailed information to Labcyte representatives Warning Warning messages report unexpected events These events may not cause a functional or operational problem since the Echo liquid handler can recover from most situations that create a warning Info Info messages report the normal activity of the Echo liquid handler Error Error messages a...

Page 160: ...5 60 Echo Liquid Handler User Manual ...

Page 161: ...bars and destination plate gripper grid Six month preventive maintenance is provided by Labcyte field service representatives if the Echo liquid handler is still under warranty or under a service contract All user maintenance procedures are provided in this chapter This chapter contains the following sections Maintenance schedule Daily maintenance Echo maintenance alerts Scheduled maintenance Main...

Page 162: ...fluid bottle level daily If the coupling fluid level is at 350 mL or less clean and refill the coupling fluid bottle and add algaecide See Refill the coupling fluid bottle on page 6 8 If the coupling fluid is cloudy clean and refill the coupling fluid bottle and add algaecide See Clean and refill the coupling fluid bottle on page 6 10 Check the waste bottle daily older Echo systems only Empty it i...

Page 163: ...g fluid temperature is outside the normal range 21 2 C 23 0 C a yellow warning light appears on the Echo front and back panels If the coupling fluid temperature continues to rise or fall outside of the warning range 20 2 C 24 0 C a red fault light appears When you see the warning or fault lights click the Diagnostics tab in the Echo software to view the specific alert Figure 6 1 Diagnostic window ...

Page 164: ...rmal range 21 2 C 23 0 C Check the chiller setting it should be at 22 C See Coupling fluid temperature and indicator on page 5 36 The waste bottle is full older Echo systems only Empty the waste water bottle See Empty and clean the waste water bottle on page 6 13 Fault light is red The user supplied compressed air supply is low 70 PSI or nonexistent Increase air pressure See Compressed air pressur...

Page 165: ...ing and faster depletion of coupling fluid Empty and clean the waste water bottle on older Echo systems only Note The waste water bottle should contain very little if any water If you are frequently emptying the waste water bottle contact Labcyte Service and Support Once per month Once per month the Echo user should perform the following procedure Clean the anti static bar emitter points and groun...

Page 166: ...th preventive maintenance Please talk to your Labcyte service representative or call Labcyte Service and Support to schedule the date If you are no longer in warranty or have a service contract contact Labcyte to obtain a quote and schedule preventive maintenance See Contact information on page 7 1 ...

Page 167: ...re required during Echo maintenance procedures Metric Allen wrenches 2 5 mm 3 mm and 4 mm Phillips screwdriver 2 Long needle nose pliers Part retrieval tool claw style retractable spring Lint free wipes Penlight flashlight Dental mirror Shop cloth to clean up spilled liquids Compressed air or vacuum with brush Materials required The following materials are required during Echo maintenance procedur...

Page 168: ...luid has dropped to the refill required level Refer to the Labcyte Echo Integration Manual Materials required Distilled water sufficient to fill the 1L bottle to 900 mL Caution Do not use de ionized water as it may corrode the fluidics system 50 μL one drop algaecide Labcyte catalog number ECHO AL04 Gloves for protection when handling the algaecide Paper towels or shop cloths to deal with any unex...

Page 169: ...efully inspect the coupler fluid tube and attached flexible tubing for the presence of bubbles If you see bubbles you will need to purge then recycle the fluid through the system after you complete this procedure See Purge the fluid circuit on page 6 11 and Cycle fluid through the Echo system on page 6 13 8 Reconnect the vacuum line and turn the coupling fluid pump on through the DIAGNOSTIC window...

Page 170: ...ail Tools and materials needed 4 liters of distilled water The Echo fluidic system holds about 1100 mL of fluid approximately 225 mL in the fluidic circuit and 875 mL in the coupling fluid bottle Emptying purging and filling the system will be performed several times Caution Do not use de ionized water as it may corrode the fluidics system An empty clean 1000 mL coupling fluid bottle square bottle...

Page 171: ... coupling fluid bottle cap 4 Remove the coupling fluid bottle from its caddy and empty the bottle 5 Replace the cap and its attached tubing onto the empty coupling fluid bottle Return the coupling fluid bottle to its caddy Purge the fluid circuit 1 Turn off the coupling fluid pump 2 In the Diagnostic window of the Echo software select the reverse coupling fluid pump direction COUPLING PUMP DIRECTI...

Page 172: ...2 Inspect the bottle bottle cap and seal ring for cracks other damage or foreign material that would cause a vacuum leak Replace if necessary 3 Fill the coupling fluid bottle with 900 mL of distilled water and add 1 drop 50 μL of algaecide Note The ratio of algaecide to water is optimized for deterring the growth of algae while minimizing residual moisture on the bottom of the plate 4 Return the c...

Page 173: ...ed empty fill cycles have been performed remember to add algaecide at the ratio of 50 μL per 1000 mL of distilled water 5 Turn on the coupling fluid pump 6 Check the COUPLING FLUID status on the Echo software DIAGNOSTICS window The indicator should be green Note If the empty purge fill procedure is being performed more than once add the algaecide only during the last fill of the coupling fluid bot...

Page 174: ...and tube a Add Alconox or hand soap and water to the bottle b Scrub the bottle and tube with a plastic bristle bottle brush c Thoroughly rinse the bottle and tube to ensure there is no soapy residue Use distilled water for the final rinse Note If you see foam while refilling the bottle rinse the bottle and tube again with hot tap water 5 Inspect the bottle bottle cap and seal ring for cracks other...

Page 175: ...h maintenance is due replace it immediately Figure 6 4 Water filter in good condition versus discolored Tools and materials needed Inline water filter P N 001 4561 Nylon hose clamps 11 32 in if needed P N 001 4045 Compressed air or vacuum with brush To replace the water filter 1 Turn off the coupling fluid pump from the Diagnostics window 2 Remove the hose clamps from the ends of the water filter ...

Page 176: ...well Periodic cleaning will maintain proper anti static bar operation Once a month dry brush the anti static bars using the following procedure 1 Turn off the Echo carriage motors by pushing the Emergency Off switch 2 Open the process door by clicking Door Open in the Diagnostic window 3 Turn off the anti static bars by clicking Anti Static Bars On in the Diagnostic window 4 Locate the two anti st...

Page 177: ...nspected for particles and cleaned monthly Tool needed Compressed air or vacuum with brush To clean the destination plate gripper grid 1 Display the Diagnostics window in the Echo software 2 Click Door Open to open the process door 3 Click Destination Plate Out The software extends the destination plate gripper and automatically turns off the electrical current Figure 6 8 Destination plate gripper...

Page 178: ...C power switch if it is not already off 2 Unplug the AC cord from the Echo power entry module 3 Pry open the power entry module fuse cover with a small screwdriver 4 Extract the fuse holder with a small screwdriver 5 Replace the two AC power fuses See Table 6 3 Figure 6 9 Replacing the AC power fuses 6 Return the fuse holder to the power entry module 7 Close the fuse cover 8 Plug in the power cabl...

Page 179: ... the following options If the Echo liquid handler remains connected to the power source chiller air and vacuum lines then perform the maintenance procedure described in section 6 3 1 If the Echo liquid handler will be disconnected and stored for an indefinite period of time then the instrument must be prepared for storage See section 6 3 2 6 3 1 Maintenance During Extended Non Use The following pr...

Page 180: ... keep the acoustic transducer wet during storage When you are ready to return the Echo to full operation contact Labcyte Service and Support The Labcyte service engineer will determine what assembly or testing steps are required to ensure that the instrument is operating correctly Note If you plan to store the Echo liquid handler in a different facility follow the moving procedures in the section ...

Page 181: ... alert information 7 1 Contact information If you need help in the following areas Technical support Applications support Service Repair Parts supplies and accessories Please contact Labcyte Technical support line 408 747 2000 option 8 or 408 542 7866 Email support Labcyte com Address Labcyte Inc 1190 Borregas Ave Sunnyvale California 94089 877 742 6548 408 747 2000 ...

Page 182: ...er has been turned on but it 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 computer and the Echo liquid handler The Echo software is not configured IP address or hostname to connect to the instrument The Echo liquid handler...

Page 183: ...e chiller setting it should be at 22 C See Coupling fluid temperature and indicator on page 5 36 The waste water bottle is full on older systems only See Waste bottle indicator on page 5 35 The process door is neither fully open nor fully closed Check for obstruction Check compressed air pressure it could be too low See Compressed air pressure indicator on page 5 35 The upper anti static bar is ne...

Page 184: ... Motion Off switch has been activated See EMO indicator on page 5 34 A coupling fluid leak has been detected See Fluid leak indicator on page 5 34 LCD message indicates user assistance needed Follow instructions on the LCD screen Client PC indicates a problem This message is generated by custom programming See your System Administrator Table 7 1 Echo Maintenance Alerts Maintenance Alert Procedure ...

Page 185: ...ill maintained the high intensity of earlier devices These devices created drops with a continuous application of acoustic energy to form geysers of small droplets or patterns of disturbances on the liquid surface where some of the swells grew large enough to pinch off and become drops The introduction of a lower intensity process that both focused and pulsed the acoustic energy to create a single...

Page 186: ...l ADE All Echo liquid handlers are pre calibrated to optimize these parameters for each fluid type Source microplate material and characteristics A transfer protocol s source microplate is specified in the transfer protocol definition from a list of Labcyte validated and predefined source plates Well fluid fill height The fluid fill height in a microplate well is measured by the Echo liquid handle...

Page 187: ...lses used during the source plate survey does not eject droplets of fluid it measures the fluid composition impedance and calculates the well fill height for each individual source well If the fluid is very stable the user can go to the Transfer Option in the Advanced tab to schedule surveys to be performed less often after a designated number of runs for quicker transfer throughput A source micro...

Page 188: ...ucer and source microplate well The acoustic transducer assembly includes a fixed focal length lens In Figure A 1 below two adjacent wells with different fluid fill heights are shown Before a droplet can be ejected the acoustic transducer must be positioned vertically on its Z axis to correctly focus the acoustic energy near the fluid s meniscus Figure A 1 Fixed focal length dictates Z axis adjust...

Page 189: ...ing fluid is responsible for Maintaining a stable acoustic transducer temperature Transducer performance is not consistent if its temperature changes during use Efficiently coupling the acoustic energy between the transducer lens and the bottom of the source microplate If the acoustic energy were to travel through air it would be substantially attenuated and the Echo acoustic droplet ejection woul...

Page 190: ...being ejected from a volume of water The acoustic energy pulse creates a mound of fluid at the surface which is pushed up A droplet forms on a neck and eventually pinches off Its momentum carries it up to a receiving surface while the fluid mound recedes into the larger fluid volume Figure A 3 A droplet ejection ...

Page 191: ...plet is traveling upward at approximately 1 m s to a glass slide In each of the four images the droplet has been photographically captured at six points in time by a multi flash strobe at intervals of 400 μs The four images show 1 20 40 and 100 drops deposited on the slide Figure A 4 Multi stroboscopic image view of ADE ...

Page 192: ...A 8 Echo Liquid Handler User Manual ...

Page 193: ... Note You can use either the front scanner or the side scanner but not both The load registration pins on the source and destination microplate assemblies obstruct the barcode scanners from seeing all of a microplate s front and side surfaces The obstructed areas are different depending on the microplate s flange height Refer to the ANSI SBS 3 2004 Bottom Outside Flange Dimensions specification fo...

Page 194: ...de of a microplate cannot be used for a barcode label Figure B 1 Short flange height barcode locations and obstructed regions B 2 Medium flange height microplates Left side Both source and destination microplates are obstructed on their left sides The 1 52 cm 0 6 in obstructed area is shown in Figure B 2 Front side There is no obstruction on the front side of either source or destination microplat...

Page 195: ... coupling fluid that is circulated through the Echo system Its solid state design is compact environmentally friendly energy efficient and designed for long life and ease of use Your Labcyte service representative will set up the chiller with the Echo system Refer to this section for safety information equipment setup if the Echo system is relocated operation and troubleshooting Figure C 1 Labcyte...

Page 196: ...his section during installation relocation maintenance and normal operation of a chiller Warnings Warning Do not use the chiller if the power cord is frayed This condition can cause a shock hazard Warning Do not disassemble the chiller Access to internal components can cause a shock hazard Improper servicing can damage the chiller unit and void the warranty Cautions Caution Do not operate the chil...

Page 197: ...ached to the fluid panel see Echo liquid handler user manual 1 Place the chiller on the benchtop near the Echo unit or on the floor under the Echo unit 2 Connect the Echo tubing assembly to the fluid connections on the chiller labeled Coolant Return for liquid returning to the chiller and Coolant Supply for fluid exiting the chiller Figure C 2 Tubing assembly connections 3 Connect the power supply...

Page 198: ...abcyte chiller has two system alarms that are shown on the chiller display RTD OPEN or RTD SHORT FAN FAIL RTD OPEN or RTD SHORT The connector has come loose or the temperature sensor has failed Turn off the chiller reconnect the cable and turn it back on If the alarm persists turn off and unplug the chiller and then contact Labcyte Service and Support See Contact information on page 7 1 FAN FAIL T...

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Page 200: ...d Handler User Manual Labcyte Part Number 001 5331 Labcyte Inc 1190 Borregas Avenue Sunnyvale CA 94089 1302 Telephone 1 408 747 2000 Toll free USA 1 877 742 6548 Fax 1 408 747 2010 email support labcyte com www labcyte com ...

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