1 - GE Healthcare Life Sciences
Contents
1. 6 1 3 Power requirements Mains voltage 100 120 V or 220 240 V 10 50 60Hz single phase Power consumption 200 VA 6 1 4 Media supply The Ettan Spot Picker requires a supply of deionized or distilled water Milli Q or the used wash destain solution 6 2 Moving the instrument WARNING The Ettan Spot Picker instrument weighs about 41 kg Two persons are required to lift the instrument CAUTION Do not use the Y arm as a handle when moving the Spot Picker Always lift the instrument from the base To move the system to another room 1 Shut down the Ettan Spot Picker and the PC 2 Disconnect the communication cables and the frame grabber cable at the rear panels on the instrument and the PC 3 Disconnect both mains supply cords from the wall outlets the instrument and the PC 4 Disconnect the monitor cable and the monitor mains supply cord at the PC and the monitor 48 Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA Moving and reinstalling the system 6 5 To prevent mechanical damage install the armlock securing the Y arm e mount the tensioning screw that immobilizes the Y arm e mount the two securing screws that hold the armlock in place Armlock Y arm Tensioning Securing screw crews 6 Carefully place the instrument the PC and the monitor on a trolley CAUTION Lift the instrument at the base Do not use the X or Y arms as handles 7 Move the system to the new location2. Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA 49 6 Moving and reinstalling the system 6 3 Reinstalling the Ettan Spot Picker 1 Selecta place for the Spot Picker where it is possible to access the mains power switch on the rear panel of the instrument 2 Remove the armlock 3 If the Z arm has been removed reinstall in its highest position If you need to reinstall the Ettan Spot Picker Instrument Control Software see instructions in Section 6 4 6 3 1 Electrical connections Reconnect the cables according to the diagram in Fig 6 1 PC Monitor Framegrabber COM 1 Serial A Ettan Spot Picker Output 24 V DC im To mains outlet MAINS RS 232 To mains outlet Network To mains outlet Fig 6 1 Electrical connectors WARNING Use only the power cords and cables supplied with the Ettan Spot Picker system Use of incorrect power cords can cause damage to the instrument Use of damaged power cords can cause injury 50 Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA Moving and reinstalling the system 6 Connecting to mains power WARNING If an Ettan Spot Picker configured for 100 120 V is connected to 220 240 V the instrument can be severely damaged Make sure that the correct fuse drawer for the voltage range is installed During the original installation the Ettan Spot Picker is configured for one voltage range by installing the appropriate fu
3. Maintenance EN Click Place Calibration Marker the picker head moves to the calibration position and down towards the tray The picker head stops at the Z coordinate shown under Calibration Marker the default Z height is 100 which will be above the tray The picker head immediately returns and the video camera is moved attempting to locate the calibration foot 8 Change the Z Coordinate stepwise to lower values and click on Place amp Calibration Marker again Repeat until the calibration foot is successfully attached onto the tape The final Z coordinate is normally 65 70 Note Check before each step that the remaining distance is not smaller than the change of Z coordinate 9 When the calibration foot has been placed onto the tape and then identified by the camera the frame around the detection window changes from red to green Camera Calibration Start Position X 100 250 Y 100 250 fi 50 0 fi 50 0 Maye Intalize Instrument r Calibration Marker Z Coordinate 30 150 ES Place Calibration Marker Temporary Calibration Values Cam X Distance 5 40 Ips OO a rea ed pr Cam Y Distance 20 60 48 60 Calibration marker found Step 4 Click Place Calibration Marker to put the marker down Adjust Z position until i 3939 the marker is placed successfully If the marker is not found adjust the iii anco MS Temporary calibration values manually lt Back Next gt
4. Camera Y Distance 2 lt Set Contrast gt lt Set Threshold gt New Calibration Values Cancel Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA 39 ED Maintenance 40 10 Repeat the procedure four times When the calibration has been successfully performed click on Save to save the new calibration values Camera Calibration Start Position x 100 250 Y 100 250 150 0 150 0 Move Rielle Instrument Calibration Marker Z Coordinate 30 150 les Hice Walibratiar MeTker lt Set Contrast gt lt Set Threshold gt Temporary Calibration Values Cam X Distance 5 40 2400 DE DA II Cam Y Distance 20 60 48 60 Ready A Im Step 6 New Calibration Values The offset marker has been detected 5 times Click Save to save the new l d close this window EE 409 ET values and close this window EES Camera Y Distance Save Cancel sae __ Cancel Troubleshooting the camera calibration procedure If the calibration foot is not visible in the camera view window although it is underneath the camera adjust the aperture until the calibration foot is visible If the calibration foot is visible in the camera view window but not visible in the detection window adjust the Contrast and Threshold sliding bars until an image is visible in the detection window If the cali
5. the instrument immediately ceases movement and all motors are de energized WARNING To prevent fire or shock hazard do not spill liquids into the camera body Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA Safety instructions E TEE 2 3 Other warnings WARNING HEAVY OBJECT The Ettan Spot Picker weighs about 40 Kg Two persons are required to lift the instrument WARNING NO SERVICEABLE PARTS INSIDE Do not open covers Service and maintenance should be performed by qualified personnel Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA 15 2 Safety instructions 16 2 4 Emergency procedures 2 4 1 Emergency shutdown In the event of an emergency press the Stop button on the Ettan Spot Picker front panel The instrument immediately ceases movement and all motors are de energized 2 4 2 Power failure routine In the event of a power failure the experiment is interrupted in an undefined state However the data for gel plugs picked up to the power interruption is saved When power returns this will happen 1 The picker head and the syringe pump dilutor are homed 2 The PC starts and displays the log in dialogue 2 4 3 Restart procedure In the event of system shutdown due to power failure emergency stop or process interruption malfunctions must be rectified before the Ettan Spot Picker is restarted To restart the Ettan Spot Picker 1 Check that the Power indicator on the
6. 0A 220 240V 1 0A 50 60 HZ Fig 1 2 Location and layout of rating labels 10 Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA Introduction ET 1 2 Package contents The Ettan Spot Picker instrument is delivered with the products listed in Table 1 3 Picker heads can be ordered separately see ordering information in Chapter 7 LETTIE TE LETTIE NN EE EEN TEEN EE EE We ET Glass lid for gel tray 1 Inlet tubing package Waste bottle and tubing Microplate rack RS 232 cable Accessories for camera calibration camera calibration foot 5 white sheet for camera calibration and click and pick function 2 Table 1 3 Ettan Spot Picker delivered products 1 3 Accessories In addition to the items delivered with the system Ettan Spot Picker requires the following accessories e image scanner Image Scanner or Typhoon e gel image evaluation software e g ImageMaster 2D Elite e 96 well microplates See Chapter 7 for specifications and ordering information If you want to use other scanners please contact Amersham Biosciences Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA 11 1 Introduction 12 1 4 The Instrument Handbook This handbook provides safety instructions technical information and basic operating instructions for the Ettan Spot Picker instrument In addition maintenance schedules and instructions for user maintenance are includ
7. 1147 62 Edition AA 67 Reference information 68 1 3 2 Related products Table 7 14 list products that are related to Ettan Spot Picker these products can be ordered from Amersham Biosciences Table 7 14 Products related to Ettan Spot Picker For code numbers marked with an asterisk see www amershambiosciences com Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA Index A AECESEDINES A cheese nepal Bots attaching a picker head DENCH Space Lima buttons front panel _ C calibratiing the camera CAMETA ui calibration us calibration procedure focus adjustment camera calibration troubleshooting changing picker head cleaning after spot picking regulai rieti communications settings connecting to the PC CONNECTIONS seriali control software reinstalling accio delivered products diagnostic guide E electrical connections Filiali Environment eec cesses Ettan Spot Picker instrument Ettan Spot Picker system F focusing the camera _ frame grabber card WOME Parel it fuse Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA 69 AAA A OE RE DE DE EE 41 G Ee EE O RA A 18 60 es oi ME IR OE EE ana 19 H HOME AA EA EE N EE N 22 indicators ronkpanel EE GR Ee e
8. Ettan Spot Picker instrument lights 2 Select Programs Ettan Spot Picker Ettan Spot Picker from the Windows Start menu 3 Reload the result file and continue picking see instructions in Ettan Spot Picker User Manual Chapter 6 Section Resume picking after Ettan Spot Picker has been stopped Note The last picked gel plug may be lost This depends on the exact moment when the interruption occurred Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA Basic Operation E 3 Basic Operation This chapter provides basic operating instructions for the Ettan Spot Picker instrument including starting stopping and preparing for gel loading Refer to the Ettan Spot Picker User Manual for instructions on picking gels It is assumed that the instrument and the PC have been correctly installed by Amersham Biosciences personnel 3 1 Preparing the instrument CAUTION Cleanliness is very important for subsequent analysis of the gel plugs Always wear protective gloves when handling gel and microplates WARNING The picker head and camera assembly can make sudden rapid movements Make sure that power to the Spot Picker is disconnected while working with the gel tray racks and picker head 3 1 1 Tubing Before starting a run a liquid supply of double distilled water or the wash destain buffer must be connected to the picker head via the Plumbing connec syringe pump The waste tube from the rinse station should be ton
9. procedure CAUTION Before starting camera calibration make sure that the Z arm is properly mounted as high as possible on the Y arm In the main application window under Load Pick List select marker type Black Markers Camera Calibration UM i ea e C sgi i i zz oa Calibration CEI Position x 100 250 Y 100 250 150 0 Calibration Marker Z Coordinate 30 150 fi 100 Temporary Calibration Values New Calibration Values Camera X Distance Camera Y Distance Intalee Instrument Place calibration arken Cam X Distance 5 40 24 03 Cam Y Distance 20 60 E 63 2 Select System Camera Calibration lt Set Contrast gt lt Set Threshold gt Pe Ready Move the picker head to an appropriate position using the Move button The camera will be centered at the position where the calibration marker will be placed lt Back Next gt Canes 3 Click on Move to move the camera to the position where the calibration foot will be placed X 150 Y 150 4 Place the calibration foot onto the picker head Locate the calibration position by placing an object under the camera or drawing a line with a pencil View the left window of the Camera Calibration screen for orientation Place a strip of double sided adhesive tape across the calibration position Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA
10. removing the picker head ooccconncoccconcnoronoacccaninaononcccnorononnns 35 rinse station setting position Of ee ee ee ee EE ee 27 Boks 2 CONNECCION lia DI safety les AE EE OE eo 14 DIEF ISIS me be ES O e EE EN 52 Site requirements lai 47 software PPP E OO E 53 SCC HO aaa 65 SIE aie iS e E 65 etlike sib ee 65 starting Ettan Spot Picker ooccccoonccconnnoncnnnnnnnoncnnnnrononcnnoncnnnnnnos 2 WII Lita 25 system setup SCreen siriani 25 T HOC lied SPECIAL aras 65 troubleshooting guide sisi i iit 43 UDC an io 17 V Videa Camela EE EE EE EE EE EE ER EG teed 64 W WasiVdestal SOLUTION usa iia on 48 EE N AA 48 rr EE eg Amersham Biosciences a Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA
11. to other products recommended or described in this manual and used in the same state as it was delivered from Amersham Biosciences except for alterations described in this manual All goods and services are sold subject to the terms and conditions of sale of the company within the Amersham Biosciences group which supplies them A copy of these terms and conditions is available on request Ettan ImageMaster ImageScanner PlusOne GelBond and Typhoon are trademarks of Amersham Biosciences Limited Amersham and Amersham Biosciences are trademarks of Amersham plc Gilson is a trademark of Gilson Inc Coomassie is a trademark of Imperial Chemical Industries PLC Decon is a trademark of Decon Laboratories Limited SYPRO is a trademark of Molecular Probes Inc Matrox is a trademark of Matrox Electronics Systems Ltd Microsoft and Windows NT are trademarks of Microsoft Corp Sony is a trademark of Sony corporation Amersham Biosciences AB 2000 All rights reserved Office Addresses Amersham Biosciences AB SE 751 84 Uppsala Sweden Amersham Biosciences UK Limited Amersham Place Little Chalfont Buckinghamshire England HP7 9NA Amersham Biosciences Inc 800 Centennial Avenue P O Box 1327 Piscataway N J 08855 1327 USA Amersham Biosciences Europe GmbH Munzinger Strasse 9 D 79111 Freiburg Germany Amersham Biosciences KK Sanken Building 3 25 1 Hyakunincho Shinjuku ku Tokyo Japan Et
12. 2 Edition AA 51 6 Moving and reinstalling the system 32 6 3 2 Power up check To verify the function of the Spot Picker at the new location 1 Switch on the mains power at the rear of the instrument 2 Check that the Power indicator lights 3 Switch on the mains power to the PC 4 To start the Ettan Spot Picker Instrument Control Software Select Programs Ettan Spot Picker Ettan Spot Picker from the Windows Start menu fo Ettan Spot Picker d Cj Ettan Spot Picker 5 Click on OK in the Homing screen to start the homing procedure where the picker head moves to the home position 6 3 3 Camera calibration Perform a camera calibration see Section 4 4 6 3 4 Shutdown To shut the Ettan Spot Picker instrument down 1 Close down the Ettan Spot Picker software by choosing File Exit 2 Switch off mains power on the Ettan Spot Picker rear panel Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA Moving and reinstalling the system 6 E 64 Reinstalling the Ettan Spot Picker Instrument Control Software IMPORTANT Before installing the software back up your result files To reinstall the control software run the installation program setup exe from the Ettan Spot Picker CD ROM and follow the instructions on the screen The installation program installs the software in the following order 1 Instrument software 2 Frame grabber software 3 Ettan Spot Picker software The PC must be restarted after each ste
13. 8 30 C 65 86 F Weight 41 kg GelBond PAG film backing Table 7 10 Ettan Spot Picker technical specification 1 2 2 Performance specification Picking efficiency gt 99 of spots transferred from gel to microplate Picking capacity 9600 protein spots per pick list Process capacity 96 samples in less than 30 minutes Table 7 11 Ettan Spot Picker performance specification Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA 65 Reference information 66 7 2 3 Regulatory requirements The Ettan Spot Picker system meets or exceeds the standards listed in Table 7 12 Safety UL 1260 UL 1492 camera CSA C22 2 No 151 IEC 1010 1 EMC EN 50082 1 EN 50022 frame grabber FCC Class A EN 50081 1 EN 50024 frame grabber Table 7 12 Regulatory requirements Spot Picker instrument Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA Reference information TEE 73 Ordering information Ettan Spot Picker including control software picker heads and reference markers is ordered under Code No 18 1145 28 The delivery package is detailed in Section 1 2 7 3 1 Accessories Table 7 13 lists the accessories that can be ordered from Amersham Biosciences LN E EU Camera calibration foot calibration foot Camera calibration foot 18 1143 33 EN GN EE click and pick function LEEN ON EE Table 7 13 Accessories for Ettan Spot Picker Ettan Spot Picker Instrument Handbook 18
14. Cleaning procedures a cio 33 4 2 1 Cleaning the instrument regularly ee see ii 33 422 Cleaning the instrument after spot DICKINB ee ee ee ee 34 Contents VI 4 55 OMANI PiEkEeriead ent is 35 45 Removing the picker head ss in riali fia 30 43 2 Attaching a new picker head ese se ee ee ee ee ee Ee 36 AA Callbratingthe Camera sutil 37 4 4 1 Required accessories ti 37 4 4 2 FOCUSING TIE IONO EA AAA EE CIA 4 4 3 calorada DIOCECOUIG vont set ke RR OO 38 4 5 Replacing the MAINS TUSE usina 41 4 6 Preventive maintenance sk ida tal GE 42 Troubleshooting TNA DE Mas E EE ER ON AE OR o 44 E A O O N EE De n 46 Moving and reinstalling the system 6 1 Me Te QUIENES aan 47 6 1 BenNGh SPICE Sraa OE NR ID 47 OAZ ENVIO ERA ME E 4 Ds P werreguiremernlS follia 48 6 1 4 MENS ODO iatale OE de GR iS 48 6 2 MOVINStNe RSTOMeni SEE EED GE eee ee 48 6 3 Reinstalling the Ettan Spot Picker iii nre 50 6 3 1 Fie liEal CONMECHONS Ad 50 03 7 MED 52 6 35 BE CQ OV QI OI RE TE OE EE N N 52 6 3 4 OO ea OE IE ON EE N 52 6 4 Reinstalling the Ettan Spot Picker Instrument Control Software 53 6 4 1 LFISTPUITICNT SOWIE is DEE lei 54 6 4 2 Frame grabber SO WATE ccccccecccceccccesecceseccecececaneecanseeenenes 55 6 4 3 Ellan Spot EIC SOWAT aa 58 Reference information Folks da da aa a lentes tees Me het 59 Ab CONC nadal nadia 60 PZ CON lle gini ro lr ail 62 LIS WE Keo 63 7 1 4 else ad EE ER ON ED 64 La SS DCCIICAUOWS ae diria oe Ee A Me
15. Eee nit 1D selector Q 2 Fig 7 6 Rear panel with mains power connector 62 Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA Reference information AE 71 3 The picker head The picker head is mounted on the Z arm just behind the video camera During a run the picker head is lowered into the gel and on to the gel backing The punch excises a gel plug by sideways movements jazz and the gel plug is then aspirated into the picker head The aspiration is achieved by the syringe pump via the liquid column in the tubing After moving to the microplate rack the picker head dispenses the gel plug into a microplate well Before the next pick the picker head moves to the rinse station where it is rinsed by aspirating and dispensing liquid Tube fitting Piston Retaining spring Punch Fig 7 7 The picker head The retaining spring creates a pressure against the gel backing during picking thus ensuring that the gel plug will be properly excised When the Z height is correctly adjusted the picker head should be lowered until a gap is formed between the piston and the body of the picker head thereby allowing the spring to press against the gel backing Fig 7 8 Spring action when lowering the picker head Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA 63 Reference information 7 1 4 Video camera The video camer
16. Fuse Fuse drawer for 220 240 voltage selection 5 Remove the old fuse s and insert the new fuse s 6 Insert the fuse drawer into its receptacle on the rear panel Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA 41 ED Maintenance 4 6 Preventive maintenance 42 Table 4 5 lists the preventive maintenance PM operations that should be performed by qualified service personnel during the yearly service visit Ettan Spot Picker system Overall inspection and cleaning Connections Check that all cables are properly connected and undamaged Homing phase Check function necessary Log file Check for errors that may require action Table 4 5 Preventive maintenance schedule Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA Troubleshooting 5 9 Troubleshooting This chapter provides basic diagnostic and troubleshooting guides The diagnostic guide is based on the error messages that are displayed by the Ettan Spot Picker Instrument Control Software see Table 5 1 The basic troubleshooting guide focuses on error symptoms related to instrument operation see Table 5 2 For errors related to gel picking refer to the Troubleshooting chapter in the Ettan Spot Picker User Manual Fig 5 1 shows a typical error message A short explanation is displayed by clicking on Details gt gt gt ERROR 004 ES Move failed Details gt gt gt MA Eo ES ERROR 004 Move failed Failed to move pi
17. GB manual proteomics Ettan Spot Picker Instrument Handbook Amersham um 18 1147 62 e Y Biosciences Important user information All users must read this entire manual to fully understand the safe use of Ettan M Spot Picker WARNING A The Warning sign highlights an instruction that must be strictly followed in order to avoid personal injury Be sure not to proceed until the instructions are clearly understood and all stated conditions are met Caution The Caution sign is used to call attention to instructions or conditions that must be followed to avoid damage to the product or other equipment Be sure not to proceed until the instructions are clearly understood and all stated conditions are met Declaration of conformity Safety standards This product meets the requirements of the Low Voltage Directive 72 23 EEC through the harmonized standard EN 61 010 1 1993 A2 1995 EMC standards This product meets the requirements of the EMC Directive 89 336 EEC through the harmonized standards EN 50081 1 and EN 50082 1 WARNING This is a Class A product In a domestic environment this product may cause radio interference in which case the user may be required to take adequate measures The CE symbol and corresponding declaration of conformity is valid for the instrument when it is used as a stand alone unit or connected to other CE marked Amersham Biosciences instruments or connected
18. a is used to establish the coordinates for the gel to be picked By detecting the reference markers on the gel and comparing with the coordinates in the pick list the picking coordinates can be determined Since the camera lens is located at a distance from the picker head its position relative to the picker head has to be precisely defined to ensure high precision picking The camera calibration routine provides the means for this see Section 4 4 The operator needs to adjust focus and aperture for optimal imaging Normally an aperture of 8 is used Brightness and contrast can be adjusted from the control software The lamps surrounding the camera lens provide the required lighting for reference marker detection and camera calibration Camera module Aperture Focus Fig 7 9 The video camera The video signal is processed by a frame grabber card in the PC before displaying the image in the Ettan Spot Picker Instrument Control Software 64 Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA Reference information TEE 7 2 Specifications Basic specifications for the instrument can be found in the Gilson 215 Liquid Handler User s Guide This section lists additional data for the Ettan Spot Picker 1 2 1 Technical specification Mains voltage 100 120 V 410 50 60 Hz or 220 240 V 410 50 Hz single phase Installation category 11 Mains fuse 1x T2 5 A 110 V or 2x T2 5 A 220 V Operating temperature 1
19. ad is just above the rinse station adjust the X Y position so that it is centrally placed Lower the picker head gently into the rinse station 6 Ifa gap is formed at the top of the picker head piston then the picker head has been lowered too far Raise the picker head slightly so that the gap is no longer present plus an additional 2 mm OK raise the picker head 2 mm from this position Too far Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA 27 ED Basic Operation AAA 7 Press the Set Rinse X Y Z button Rinse Station Position 2 coordinate 1535 Y coordinate EE coordinate 126 8 Set Rinse sr 3 5 4 Setting the positions of the microplates The microplates are numbered as illustrated in the System Setup screen They should be placed with the A1 well facing the operator During System Setup only this well has to be defined for each plate Fig 3 11 The numbering of the microplates To set the positions of the microplates 1 Using the Move X Y arrow move the picker head until it is centrally placed above the A1 well of microplate 1 2 Using the Move Z arrow slowly lower the picker head into well A1 28 Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA Basic Operation E E 3 The current position of the picker head is continuously read back from the instrument and displayed in the corresponding text boxes MicroPlate Positions Well Al Co
20. bration foot is not placed on the tape decrease the Z coordinate to make the picker head move closer to the tray Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA Maintenance EN TEX 45 Replacing the mains fuse A blown fuse may indicate the existence of another problem in the instrument If the replacement fuse blows don t try others Contact your local Amersham Biosciences representative WARNING For continued protection against risk of fire replace with fuses of the same type and rating only To replace the mains fuse 1 Disconnect the mains power cord from the power outlet and from the rear panel socket 2 Locate the fuse drawer on the rear panel 3 Insert a small screwdriver into the notch next to the fuse drawer Q amp SS 25 gt 3 o mE z Qu Go i ES sir O y EDE Q sw ETTING DI hd O F d for 100 120 i 3 ollags lection Fig 4 4 Replacing the mains fuse 4 Twist the screwdriver to open and remove the fuse drawer The fuse drawer contains one 2 5 A T slow blow fuse 5 x 20 mm size for a 100 120 voltage selection It contains two 2 5 A T fuses for a 220 240 voltage selection
21. c representation of Picker head movement the Spot Picker controls System Setup 0 0 X eo Rinse Station MicroPlates Fig 3 10 The System Setup screen Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA 25 ED Basic Operation AER 35 1 Moving the picker head around It is advisable to press the Go to home button prior to setting any of the EE system locations The blue arrows on the right hand side of the screen are used to move the picker head to various positions By pushing the buttons on the arrows the head will move stepwise in the corresponding directions As long as a button is pushed the head will keep moving Move X Y move the picker head horizontally the outermost 10 1 0 1 buttons will move the head in 10 mm steps the TEE middle buttons in 1 mm steps and the innermost 5 buttons in 0 1 mm steps 1 Move Z move the picker head vertically in 5 1 and 0 1 mm 0 1 steps respectively Note When the picker head is close to the rinse station the 10 mm step button can be disabled since the rinse station is located near the horizontal limit of the Spot Picker Use the 1 mm and 0 1 mm buttons to move the picker head at this stage CAUTION Be careful when using the Move Z buttons rapidly lowering the picker head into an object may damage the picker head If the picker head hits an obstacle the motor will be shut off and an error message will be disp
22. cause may be that the program and use the Home instrument has collided while Phase Tool For details see the moving in the Y direction user manual forward backward 025 Homing failed X and Y Homing failed for the X and Y Run the Set Model 215 axes axes The cause may be that program and use the Home the instrument has collided Phase Tool For details see the while moving diagonally user manual Table 5 1 Diagnostic guide Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA 45 ED Troubleshooting TEE 52 Troubleshooting Error symptom Possible cause Corrective action No picture from the camera Camera not connected to the Check camera cable and PC reconnect Reference detection unstable Contrast not properly set Adjust the aperture setting on the camera Light not coming on Power supply from 24 V output Check power cable from light disconnected to 24 V power supply Internal fuse has blow Call your Amersham Biosciences service representative No movement from the Spot Instrument not plugged in or Check that the Spot Picker is Picker following command sent mains switch turned off connected to a wall socket and from the PC that the mains switch is in the ON 1 position The Stop button has been Press the Start button on the pressed Spot Picker RS232 cable not properly Check RS232 cable and connected ensure that it is plugged into the correct port Other problems Call your Amersham Biosciences service represe
23. ce information WARNING Never put your hands underneath the picker head during instrument operation The picker head has the capacity to puncture skin In the event of an emergency press the Stop button on the front panel WARNING MOVING PARTS The picker head and camera assembly can make sudden rapid movements Always switch off mains power before replacing parts or cleaning the instrument 4 2 Cleaning procedures 4 2 1 Cleaning the instrument regularly For proper function the Ettan Spot Picker should be kept clean and dry Chemical stains and dust should be removed 1 Remove the gel tray and the microplate racks from the locator plate 2 Wipe the instrument cover with a soft damp tissue If needed use a mild detergent to remove stains 3 Clean the gel tray and the microplate racks using a mild detergent 4 Let the instrument dry completely before use Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA 33 ED Maintenance AE 422 Cleaning the instrument after spot picking When a picking run is completed 1 Remove the gel from the gel tray and store discard as appropriate 2 If the gel is to be discarded scrape the gel off the glass plate with a plastic spacer or similar object Then put the plates into a 5 Decon solution overnight to remove any gel fragments which adhere to the plate 3 Rinse the buffer lines with distilled water using the prime syringe tool 4 Rinse the liquid out of the gel t
24. cking head Try checking instrument connections Extra information Failed to read Yz motor status Fig 5 1 Typical error message Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA 43 ED Troubleshooting TEE 5 1 Diagnostics Error message Details Possible cause Corrective action 001 Reset failed Failed to reset the instrument Try checking instrument connections 002 Homing failed Failed to home the picker Try checking instrument head connections 003 Homing failed Failed to home dilutor Try checking instrument connections 004 Move failed Failed to move the picker head Try checking instrument connections 005 Move timed out Failed to move the picker Try checking instrument head connections 006 Movement out of range Movement outside travel range When picking Try changing system setup 007 Dilutor operation timed Dilutor operation failed Try checking instrument out connections 008 Dilutor operation failed Dilutor operation failed Try checking instrument connections 009 No connection Failed to read the picker head Try checking instrument position from the instrument connections and or reinitialize the instrument 010 No connection Failed to read syringe content Try checking instrument from the instrument connections and or reinitialize the instrument 011 Read version failed Failed to read instrument Try checking instrument version connections and or reinitialize the instrument 012 Read ve
25. creen to start the installation Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA 55 6 Moving and reinstalling the system AAA 3 Select No for new MIL 32 driver for Meteor II Click on Next i Meteor ll setup 56 Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA Moving and reinstalling the system Select 8 0 MB memory buffer size Click on Next Matrox DHA manager In order to be able tc grab in he Host system memory via the PCI bus bulfers mud be allocated in a physically linear memory array called non pag d memory Please select the number of megabytes KB of non peged memory to reserve fer your Matron frame grabber 1 0 ME 20MB O 4NMR Other GA The naximum size vou can allocate is 242 MB Back Cancel 6 Click OK to start copying files Matrox Imaging Products redistribution Setup le now ready to copy files to your system If you want to change any settings click Back Click Ok to begin copying files lt Back Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA 57 6 Moving and reinstalling the system E 7 When installation of the frame grabber software is finished select Yes and click on Finish to restart the PC Matrox Imaging Products Redistribuiion setup Setup has successfully installed the Matrox Imaging Product to your computer Before you can use it you must start ycur computer No wil estart me c
26. d to the PC see Chapter 6 Moving and reinstalling the system Note The Spot Picker control software will not run unless a frame grabber card is installed in the computer 2 Switch on the mains power at the rear of the instrument 3 Turn on the PC to which it is connected 4 To start the Ettan Spot Picker Instrument Control Software select Programs Ettan Spot Picker Ettan Spot Picker from the Windows Start menu fo Ettan Spot Picker E Ci Ettan Spot Ficker The picker head will be moved to the home position when the software starts For safety reasons the user must confirm the movement before starting Homing Before continuing the system must be homed Press OK to start homing Cancel 6 After homing which takes about 20 seconds the picker head is positioned as shown in Fig 3 8 22 Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA Basic Operation E Fig 3 8 Homing the picker head r Instrument Handbook 18 1147 62 Edition AA 23 ED Basic Operation 34 Software user interface When the homing procedure is ready the main application window is shown You can control the Spot Picker by using buttons in the window and by selecting functions from the pull down menus at the top of the screen Graphical representation Co ordinate of the pick list table picklist Menu bar E Ettan Spo Picker x File Syste
27. e ee 61 MOGUCU sleale 9 L locator DIATE ui dd 60 M main application WINDOW anar s 24 A Ge 62 Mans Use VO ACHING iaa de dl de ae 41 mains power connection ilatina 5 mans SWIET ladina 62 maintenance DreVerlVE sise das 42 Manuals oer aaa adi 12 Media SUPDIY lc 48 microplate TACKS silice 19 60 microplates setting positions Of ta SE ia 28 moving the instrument ets see EE in ds 48 lk De ea EO A 49 MOVINTE PICKE NCI se EES are 26 ieke Hesse hill 47 0 operating INStrUCHONS sasie ek RR n De hie se ed 17 orderne ING AO EE EE ARS 67 ACCESSO O S an ER AS A A a a 67 PelALCO LOGIC SR N a 68 P package COMETS daba 11 performance specification iia 65 pleker head EE a 214 65 lt Ie OE RR Ro RT 36 CHAMOIS RR GE A 35 OMONE dibatte 35 5 70 Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA IN n ower reduirements ee ee ee ee ee ee ee ee 48 power up check AA N riores 52 preparing the instrument o oo esse ss ee ee ee ee ee ee ee ee ee ee 17 preventive MAINTENANCE occcoccccnccoronoanroncnonorenccnnonanoccnacinoanoones 42 DION ei ii 30 R ieke or eras taa 19 60 les RE EE E EE EE EN 10 co sven 62 regular MAINTENANCE e 33 regulatory requirements ssa ans 66 reinstalling the system sesse see see ee Re ee ee Pe ee ee A 50 contro lsoftware OE N OE OE OO NN 53 frame grabber software oocoonccconcncnnnncnnnnnncnnonronennononcnnononos D5 instrument software iran 54 related DOUC Lilla lada 68
28. e the piston At the same time hold the top of the piston steady with your other hand 3 Slowly screw the picker head into the piston while holding the top of the piston still with the other hand The picker head only needs to be tightened to finger tight i Fig 4 2 Final appearance of the assembled picker head Proceed by calibrating the distance between the picker head and the camera see Section 4 4 36 Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA Maintenance EN 4 4 Calibrating the camera Aperture Focus 4 4 1 Required accessories You need the following accessories for the camera calibration e White sheet for calibration code no 18 1143 51 e Camera calibration foot code no 18 1143 33 e Double sided adhesive tape Fig 4 3 Accessories for camera calibration 4 4 2 Focusing the lens Before starting the camera calibration check that the camera lens is focused on the bottom of the tray 1 Use the Click and Pick screen for the adjustments select Tools Click n Pick 2 Place the white calibration sheet in the tray 3 Place an object on the sheet e g a piece of paper with printed text 4 With the Set Contrast and Set Brightness controls in the centre positions adjust the aperture to normal brigthness 5 Adjust the focus ring on the lens to a sharp picture Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA 37 ED Maintenance 38 1 Calibration
29. ed 1 5 Associated documentation The Ettan Spot Picker User Manual describes the experimental use of the instrument and includes descriptions of all software dialogues Gilson M 215 Liquid Handler User s Guide describes the unmodified instrument and contains technical specifications and instructions for installation and maintenance Reference will be made to instructions located in the User s Guide The Sony MBlack and White Video Camera Module XC 55 55BB Operating Instructions describes the camera The Matrox M Meteor II Installation and Hardware Reference manual describes the frame grabber board and provides installation instructions Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA Safety instructions E 2 Safety instructions IMPORTANT To avoid any risk of injury the instrument should only be operated by properly trained personnel and always in accordance with the instructions provided The purpose of this chapter is to describe safety precautions and present all safety labels that are attached to the instrument Built in safety functions are described and emergency and disposal procedures are included 2 1 Safety precautions 1 Read this entire manual before using the Ettan Spot Picker instrument 2 This instrument is designed for indoor use only 3 The instrument must always be used with the protective earth lead of the power cord correctly grounded to earth at the mains outlet 4 To per
30. imaging equipment by correcting the pixel co ordinates of the spots contained within the pick list 3 8 2 Calibration procedure The calibration works in the following way e A special calibration sheet is imaged on the scanner e The location of the spots in the subsequent image of the calibration sheet is determined e The image distortion is calculated using the differences between the known co ordinates of the calibration sheet and the measured spot locations e The image distortion data is stored as a correction table for the scanner e A gel pick list that is loaded into the Ettan Spot Picker Instrument Control Software is modified using the correction table for the particular scanner that was used when scanning the gel For detailed instructions see the documentation on the CD ROM included in the ImageScanner Geometry Calibration Kit Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA 31 Maintenance EN 4 Maintenance 4 1 Introduction Regular maintenance is important for safe and trouble free operation of the Ettan Spot Picker The user should keep the instrument clean and calibrate the camera whenever a picker head has been replaced Preventive maintenance should be performed on a yearly basis by qualified service personnel This chapter provides instructions for user maintenance and service operations and a schedule for preventive maintenance Contact your Amersham Biosciences representative for more servi
31. ins for securing microplate racks Fig 7 2 Locator plate with gel tray Two racks for microplates are placed beside the gel tray Up to four microplates can be placed in the racks at the same time However the software allows the operator to change microplates during picking enabling a large number of gel plugs to picked in one run 1 1 1 Front panel Start butt oo LED display Stop Power indicator light NE ri NOS y Fig 7 3 Indicators and controls on the front panel RE 60 Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA Reference information Power unmarked Lights when mains power is connected to the instrument and the mains switch on the rear panel is on in the 1 position yellow Lights when the Start button is pressed to home the picker head LED display yellow text Shows the current status of the Spot Picker and any error codes as they are encountered see Chapter 5 Table 7 4 Indicators on the front panel When the Start button is pressed the picker head moves to home position The yellow LED lights as long as the button is pressed The Start button can be used to home the picker head when the Ettan Spot Picker is first powered up or when the motors for the picker head have been relaxed When this button is touched the instrument immediately ceases movement and all motors are de energized When the Stop button is pressed the yellow LED in the Start button is turned off No
32. layed CAUTION Be careful when using the Move X Y buttons if the picker is in a low Z position Rapidly moving the picker head into an object may damage the picker head Always use the innermost Move X Y buttons when adjusting the sideways position at low Z heights The current X Y Z position of the picker head is continuously read back from the instrument and displayed in the corresponding text boxes Once the picker head is placed appropriately e g in the rinse station the location can be stored by pressing the appropriate Set button If the Cancel button is pressed any changes made to the System Setup will be lost To retain new System Setup parameters click on the Save amp Exit button 26 Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA Basic Operation E AAA 352 Camera calibration before picking If the picker head has been replaced the distance between the camera and the picker head needs to be calibrated before starting a picking run See instructions in Chapter 4 3 5 3 Setting the position of the rinse station 1 Using the Move X Y arrow move the picker head until it is placed above the centre of the rinse station 2 Using the Move Z arrow slowly lower the picker head into the rinse station 3 Ifit appears that the picker head will miss the centre of the rinse station adjust the X Y position of the picker head as appropriate and continue to lower the picker head 4 When the picker he
33. m Tools About File None Spot No ImageX Image PickerX PickerY Plate No Well a No Pick List Loaded Camera view n No pick list loaded m Select Marker Type white Markers C Black Markers Load Pick List Hest Move XA Load Pick List Camera controls lt Set Dentrast gt lt Set Threshold gt Initialize Instrument Go To Home Marker x Off set Marker Y Off set Adjust ural ms la a AMEN GP aa Aa At Picker head Instrument Marker detection Spot picking guide controls controls window functions Fig 3 9 The Ettan Spot Picker main application window For detailed descriptions of the Ettan Spot Picker software functions see Ettan Spot Picker User Manual 24 Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA Basic Operation E 3 5 Setting system parameters The following parameters must be set up before starting a picking run e The position of the rinse station and the microplates e The height of the gel backing material Gel Z position e The parameters used for picking the gel plugs e The acceptable area range for the reference markers in pixels This section describes the procedures for setting the positions of the rinse station and the microplates For instructions on setting parameters for gel handling see Ettan Spot Picker User Manual To set the Spot Picker parameters choose System System Setup Schemati
34. mit sufficient cooling ensure that the vents at the top and bottom of the instrument are not covered 5 Do not operate the instrument in extreme humidity above 95 Avoid condensation by letting the unit equilibrate to ambient temperature 6 Do not operate the instrument in direct sunlight 7 Keep the instrument dry and clean Wipe regularly with a soft damp tissue Let the instrument dry completely before use 8 Any equipment connected to the instrument should meet the requirements of the EN 61 010 1 or other international safety standards WARNING Do not use the Ettan Spot Picker in any other way than described in this Instrument Handbook The protection provided by the instrument may be weakened if other operations are performed Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA 13 2 Safety instructions 14 2 2 Safety labels The following safety labels are attached to the Ettan Spot Picker instrument to warn the user of potentially hazardous conditions At the picker head Fig 2 1 Safety label located on the Y arm near the picker head WARNING MOVING PARTS The picker head and camera assembly can make sudden rapid movements Keep all body parts clear when the Spot Picker is in operation WARNING Never put your hands underneath the picker head during instrument operation The picker head has the capacity to puncture skin In the event of an emergency press the Stop button on the Spot Picker
35. ng image analysis These reference markers are used to transform image pixel co ordinates into distance values for spot picking Following excision from the gel gel plugs are dispensed into 96 well microplates along with a volume of dispensing fluid 1 1 The Ettan Spot Picker system Fig 1 1 The Ettan Spot Picker system Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA 9 1 Introduction The Ettan Spot Picker system comprises the following e Ettan Spot Picker instrument e Ettan Spot Picker instrument control software running under Windows NT operating system on a PC e PC with frame grabber card for video display The package contents are described in Section 1 2 A detailed description of the Ettan Spot Picker instrument is provided in Chapter 7 of this manual 1 1 1 Rating labels A rating label is located on the rear panel of the Ettan Spot Picker instrument see Fig 1 2 Pay attention to the component ratings These ratings determine the degree of electrical hazards in the eguipment connected to the supply voltage There are however other hazards which may be more severe see Chapter 2 Safety instructions FUSE 100 120V 220 240V EE EI 250V 2 5A T 250V 2 5A T 5x20mm 5x 20mm 2 CAUTION FOR CONTINUED PROTECTION AGAINST RISK OF FIRE REPLACE ONLY WITH THE SAME TYPE AND RATING OF FUSE MAINS SHOWN IN OFF POSITION 100 120V 2
36. ngtensa ee AE Ee Ee 65 ae Technical specication EE nsec in Be SR DEER a 65 LEC Performance specification ri 65 LO Regulatory TEQUES avisa di 66 LS Orderirie IO RMALON ss AS 67 Ll ACCES SONOS aa Roa 67 LE I CIAL prady CiS oas eE id 68 Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA List of figures List of figures Fig 1 1 Fig 1 2 Fig 2 1 Fig 3 1 Fig 3 2 Fig 3 3 Fig 3 4 Fig 3 5 Fig 3 6 Fig 3 7 Fig 3 8 Fig 3 9 Fig 3 10 Fig 3 11 Fig 4 1 Fig 4 2 Fig 4 3 Fig 4 4 Fig 5 1 Fig 6 1 Fig 6 2 Fig 6 3 Fig 7 1 Fig 7 2 Fig 7 3 Fig 7 6 Fig 7 7 Fig 7 8 Fig 7 9 Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA The Ettan Spot Picker SysteM ese see sere ee ee ee ee ee ee 9 Location and layout of rating labels ee ee ee ee ee ee eek ee 10 Safety label located on the Y arm near the picker head 14 Spot Picker tubing Sel cata 18 Placing the gel tray on the locator plate ese see see ee ee ee ee 18 FOS MOMS die Be US atada 19 Fastening lie SE EG OG ee sais AAE 19 Placing a microplate rack on the locator plate 20 Installing microplates in the racks well Al marked 20 Tie DIC KET leads narra oa 2 MOMINS ie DiEker Head RE GE Se aes 23 The Ettan Spot Picker main application window 24 The System Oe LENO SEE SR spalle Ee ie Ie ie 25 The numbering of the micr
37. ntative Table 5 2 Troubleshooting guide 46 Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA Moving and reinstalling the system 6 6 Moving and reinstalling the system TheEttan Spot Picker system is installed and tested by Amersham Biosciences service personnel after delivery This chapter provides instructions for reinstallation should you need to move the instrument to another location WARNING Always shut down Ettan Spot Picker and the PC before disconnecting or connecting cables See instructions in Chapter 4 Before moving the system check that the new location fulfils the site requirements listed below 6 1 Site requirements 6 1 1 Bench space The Spot Picker should be placed on a stable and level laboratory bench Allow space for a water bottle on the left hand side of the instrument and for the computer and monitor on the right hand side Note that the Z arm can move up to 56 cm above the bench Make sure that any shelves are mounted at sufficient height Dimensions Ettan Spot Picker 91x61 x56 cm instrument W x D x H 6 1 2 Environment Temperature operating 18 30 C 65 86 F Maximum relative humidity 80 non condensing Air pressure 75 105 kPa up to 2000 m altitude The system must not be exposed to direct sunlight Dust in the atmosphere should be kept to a minimum Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA 47 6 Moving and reinstalling the system AE
38. ompurer later Wher pour computer nas restarted you can Use pour applicationwilh the Matrox Imaging Product 6 4 3 Ettan Spot Picker Software The installation procedure for the Ettan Spot Picker software is straight forward and requires no special selections For easy set up accept the suggested file location and program group Close any running application then click OK in the first screen to start the installation see Fig 6 3 dd Ettan Spot Picker v1 00 Setup ES Welcome to the Ettan Spot Picker 1 00 installation program ia Setup cannot install system Files or update shared files if they are in use Before proceeding we recommend that you close any applications you may be running Exit Setup Fig 6 3 Ettan Spot Picker software installation start screen 58 Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA Reference information 7 Reference information Front panel tes A_N F SI da N Valve Inlet tubin Syringe pump Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA 7 1 The Spot Picker Instrument The Ettan Spot Picker is a robotic system that can move the picker head in three directions X Y and Z The syringe pump allows precise handling of liquid and picked gel plugs The valve next to the syringe pump controls the flow from the liquid supply and to from the picker head The picker head is rinsed between each pick and waste liquid is dis
39. oplateS esse see ee ee ee ee ee 28 APS IC So MO EDE N N GE N Se De DiE 36 Final appearance of the assembled picker head 36 Accessories for camera calibration ee ee ee ee ee 37 Replacing the Mains USE ido 41 Typical error message sesse reo 43 Electrica Connect Sube 50 Communications settings on the Ettan Spot Picker Sd Ettan Spot Picker software installation start screen 58 fie GATES DOL PICK OTs si Me a 59 Locator pate Wit Sel ada 60 Indicators and controls on the front panel 60 Rear panel with mains power COnnEctor de ee ee 62 hie be A CI one ge n EPA 63 Spring action when lowering the picker head 63 NRO lo aio Es AA mesa mses ua eine tala S 64 vil Introduction ET 1 Introduction The Ettan Spot Picker is designed to automatically excise and transfer polyacrylamide gel plugs containing proteins from 2D gels to microplates These gel plugs are suitable for further processing prior to mass spectrometry analysis The Spot Picker is compatible with 2D gels that have been post stamed with Coomassie silver or SYPRO dyes and for pre labelled proteins Spots of interest can be evaluated and selected by using the ImageMaster 2D Elite gel image analysis software A pair of reference markers must be included within the gel and also detected duri
40. ordinate Plate Mo 1 Co ordinate 150 2 Set Plate z 4 If it appears that the picker head will miss the centre of the well adjust the X Y position of the picker head as appropriate and continue to lower the picker head When the picker head is in the A1 well adjust it so that it is centrally placed 5 The Z position of the picker head should be adjusted so that approx 2 mm of the tip is within the well This will enhance the dispensing efficiency of the spot picker 6 When the picker head is correctly positioned in the well press the Set 1 button 7 At the first microplate press the Set Plate Z button this sets the Z height for all of the microplates and does not need to be set for each plate individually CAUTION Raise the picker head before moving to the next Al well 8 Repeat steps 1 6 for the remaining microplates without pressing the Set Plate Z button 3 6 Setting picking parameters Before picking spots from a gel a number of Picking Parameters must be set as well as Gel Z Position and Marker Area Instructions for these settings can be found in the Ettan Spot Picker User Manual rr ES Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA 29 ED Basic Operation Tf 37 Priming the system Priming the syringe removes any air bubbles in the spot picker tubing and also rinses the tubing with fresh solution It is recommended that 5 10 priming strokes are used Note It is important to ensure that the
41. p For information on the instrument software and the frame grabber software read any readme files in the picker and framegrabber folders on the CD ROM During the software installation factory defaults are stored for system parameters Therefore you need to e re calibrate the camera Section 4 4 e perform a system setup Section 3 5 Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA 53 6 Moving and reinstalling the system E 641 Instrument software Before starting installation make sure that the Windows control panel D Regional Settings is set up as follows Regional Regional Settings English United States Settings i Input Locales choose your national keyboard 1 Select the appropriate COM port where the instrument communication cable is attached Click on Next COM Port Selechon Select the communications pot that wil serve the GSIOC channel Eack Cancel 2 When the instrument software is installed click on Finish to restart the PC Setup Complete Setup has finshed coming files to pour computer Before you can use the program you must restart Windows or your computer Ho wil estart me compurer later Remove any disks from their dives and then cick Finish to complete sctup 54 Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA Moving and reinstalling the system 6 E 6 4 2 Frame grabber software 1 Click on Next in the first s
42. pensed into the rinse station The waste liquid flows through the waste tube to a container placed below the Spot Picker The picker head movements are guided by a camera system The camera that is attached to the Z arm is connected to the PC via a frame grabber card Reference markers on the gel are identified using the camera This enables the pick list co ordinates from the scanned gel to be transferred to picker head positions during automated picking The operator can also pick gel plugs manually using the camera image for orientation di gs Waste tube Y arm 2 Z arm X arm mm TNN Locator plate IE Gel tray Microplate racks Fig 7 1 The Ettan Spot Picker 59 Reference information ri A gel tray is placed on the locator plate on the base of the instrument The gel to be picked is fastened in the tray using gel holders When calibrating the camera a white sheet is fastened in the tray The white sheet is also used during manual picking to enhance contrast in the camera image Rinse station Gel holders P
43. plates which are placed in two racks on the right hand side of the locator plate 1 Install the racks on the locator plate aligning the holes on the underside with the pins on the locator plate see Fig 3 5 The rack label should face the front of the Spot Picker 2 Place up to four microplates in the racks Note that well A1 should face the front of the instrument see Fig 3 6 Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA 19 ED Basic Operation Fig 3 6 Installing microplates in the racks well A1 marked 20 Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA Basic Operation E A 3 2 Selecting a picker head Picker heads are available in different sizes for different types of gel See Chapter 7 for specifications If you need to change the picker head follow the instructions in Chapter 4 Maintenance After changing the picker head a camera calibration should be performed Instructions for calibration can also be found in Chapter 4 Tube fitting Piston Retaining spring Exchangeable part with variable diameter and height Punch Fig 3 7 The picker head rr ES Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA 21 ED Basic Operation 3 3 Starting Ettan Spot Picker WARNING The picker head and camera assembly can make sudden rapid movements Keep all body parts clear when the Spot Picker is In operation 1 Check that the Spot Picker instrument is connecte
44. position of the rinse station is correctly set in the System Setup see Section 3 5 3 before priming the system 1 Check that the tube fittings at the valve are properly tightened 2 Select Tools Prime Syringe 3 In the Prime Syringe window enter the number of priming strokes and press the Prime button Prime Syringe Priming Strokes tt lb Press Prime to start Stop Prime Exit 4 The Stop button is now enabled and can be used to interrupt the priming Prime Syringe Priming Strokes H E Friming stroke 2 of 5 30 Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA Basic Operation E 3 8 Calibrating the scanner 3 8 1 Background Ettan Spot Picker uses the X Y image co ordinates of protein spots to determine the location of the gel plugs to be removed Some imaging systems have inherent imperfections which lead to a slightly distorted gel image The distortion of the image will lead to a distortion of the spot positions The picker head will move to a position which although the correct location that it has been sent to is not representative of the true location of the protein spot To correct for such distortions the ImageScanner can be calibrated using a separate software that is included in the ImageScanner Geometry Calibration Kit The ImageScanner Geometry Calibration Tool software corrects for the distortions of the gel image that are introduced by the
45. ray with distilled water and leave the gel tray to dry 34 Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA Maintenance EN 4 3 Changing picker head The Ettan Spot Picker can be used with a range of picker heads with varying diameter and height The picker heads are easily changed by a simple screw in screw out method After changing a picker head a camera calibration should be performed to establish the offset between the picker head and the camera lens The gel Z position may also have to be adjusted in the System Setup Retaining spring O ring EE Punch Fig 4 1 The picker head 4 3 1 Removing the picker head WARNING Switch off mains power to the Spot Picker before removing the picker head 1 When mains power has been switched off move the picker head to a convenient position to work in 2 Hold the top of the picking piston with one hand and with the other unscrew the picker head from the base of the piston 3 When the picker head is completely unscrewed from the base of the piston place it in a safe location until it is reguired once more Make sure that the retaining spring is also kept safe Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA 35 ED Maintenance 4 3 2 Attaching a new picker head 1 Place the retaining spring over the screw thread on the top of the picker head 2 Screw the picker head onto the base of the piston making sure that the screw thread is insid
46. rsion max flow Failed to read dilutor max flow Try checking instrument failed connections and or reinitialize the instrument 013 Read travel range failed Failed to read the picker head Try checking instrument travel range connections and or reinitialize the instrument 015 File error Could not access file or missing file 44 Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA Troubleshooting 5 ES 018 File error Trying to access output Enter a valid output directory directory which is not valid or or change attribute for write protected directory e g uncheck read only 019 File error Could not access file or Example move scanner missing file calibration file to correct directory 020 File format error Cannot recognize the file Errors in pick list e g multiple format IR1 or IR2 coordinates 021 Internal error An application error has If the problem persists please occured contact Amersham Biosciences support 022 Scanner correction failed Failed to calculate scanner Try running the scanner correction calibration program again and replace the calibration file 023 Homing failed X axis Homing failed for the X axis Run the Set Model 215 The cause may be that the program and use the Home instrument has collided while Phase Tool For details see the moving in the X direction left user manual right 024 Homing failed Y axis Homing failed for the Y axis Run the Set Model 215 The
47. s connected to a waste bottle placed beneath the instrument see Fig See Gilson 215 3 1 Liquid Handler User s Ad AAS Meen The liguid is used for aspirating gel plugs and dispensing them into the onapter wells of the microplates The picker head is rinsed in liquid at the rinse station between each pick EE Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA 17 ED Basic Operation Inlet tubing E i p DAS Y Valve gJ Liquid supply Syringe pump CL tE AH A di Waste tubing gt Fig 3 1 Spot Picker tubing setup 3 1 2 Positioning the gel tray Place the gel tray on the left hand part of the locator plate as shown in Fig 3 2 The guide feet on the tray should fit onto pins on the locator plate It is only possible to fit the gel tray in one direction with the corner mounted guide feet facing the operator see Fig 3 3 Fig 3 2 Placing the gel tray on the locator plate 18 Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA Basic Operation E Fig 3 3 Positioning the gel tray The gel to be picked is fastened in the gel tray using adjustable clamps gel holders as shown in Fig 3 4 pan ye j V ei Fig 3 4 Fastening the gel 3 1 3 Racks and microplates The gel plugs are collected in micro
48. se drawer Before reinstalling the instrument make sure that the correct fuse drawer is installed for the mains voltage at the new location If you need to change the mains voltage configuration refer to instructions in Section 4 5 for installing mains fuses To connect the Ettan Spot Picker system to mains power 1 Connect one end of the supplied mains cable to the mains inlet on the Spot Picker and the other end to a mains supply outlet with protective ground 2 Check that the mains voltage marking on the PC corresponds to the mains supply voltage 3 Connect one end of the supplied mains cable to the mains inlet on the PC and the other end to a mains supply outlet with protective ground 4 Connect the supplied monitor power cable between the mains inlet on the monitor and a mains supply outlet with protective ground Connecting to the PC Connect the supplied communication cable between the RS 232 connector on the Ettan Spot Picker and the COM1 port on the PC Check that the switches SW1 and SW2 are set as shown in Fig 6 2 ES SW1 UNIT ID 20 SETTING SW2 SETTING BAUD MODE 0 19200 GSIOC SLAVE EXTERNAL CLOCK 2 9600 19200 GSIOC SLAVE SW1 Unit ID 22 INTERNAL CLOCK 9600 1 2 3 i 2 GSIOC MASTER Baud rate 19200 f BREAK ACTIVE SW2 Mode GSIOC 8 6 9 GSIOC MASTER BREAK PASSIVE Master Fig 6 2 Communications settings on the Ettan Spot Picker Ettan Spot Picker Instrument Handbook 18 1147 6
49. tan Spot Picker Instrument Handbook 18 1147 62 Edition AA Contents Contents Introduction 1 1 The Elam Spot Picker Sy Mila 9 lib Rane le RE DE AE noaeeenis 10 1 2 Package COmentS varas 11 LS ACCESSONES stc ER ER oe EE EE RE GE DOER EER ee 11 tA Thelnsirument Hand si ot ela 12 1 5 Associated documentation ee ee ee ee ee ee ee ee ee 12 Safety instructions Zul NS EES N EG ER EE Ee DE GO 13 22 Saly aBel RE EE ER EN a Leona 14 29 ier War ES PP 15 24 A e MEE EE ee ee N i 16 Zad Emergency SIUMOOWD soia 16 2 4 2 ROME TINS TONNE an Seen i a r one Se 16 2 4 3 Restart DIOCCO Cia ci 16 Basic Operation DL voPrepatine ne instrument asi ia 17 3d LO 17 Ek FOSON THC CONT a 18 odo PACKS and microplates des 19 Sue sel ecune a pieke Head EE ai 21 3 3 Starting Ettan Spot PIEKE pretorio lare ee ee GE 22 3 4 Software user interface ipa 24 3 5 Setting system parameters EE EDE ss 25 core Moving the picker head AFOUNC cccccccccccccceccccceceececeeceneess 26 SoZ Camera calibration before picking cccccccccceeccceccceeseneeseneeees 27 Cioe Setting the position of the rinse station ee ee ee ee 27 3 5 4 Setting the positions of the microplateS ee ee ee ee 28 3 6 Setting picking parameters esse see ee ee ee ee ee ee EE ee 29 A AS EE EDE GE Se he GE EE eer eo 30 3 8 C librating Te scanner birillo Ee ie Ee Poe ee 31 EL BICI elle 31 3 8 2 Eara dese ad RA aiar Sl Maintenance AT TMP OCIA CTI OR sro 33 4 2
50. te that the Stop button is very sensitive do not touch it unnecessarily during a run Table 7 5 Buttons on the front panel Ettan Spot Picker Instrument Handbook 18 1147 62 Edition AA 61 Reference information AE 7 1 2 Connectors All connectors are located on the rear panel of the instrument see Fig 7 6 The mains switch and mains fuse drawer are part of the mains inlet connector at the lower end of the panel Es ES O 2500449 WUT WT OBN WT OAN MUN INDICATES CIRCUIT GROUND NPUT MADE IN USA RE L SW1 UNIT ID 20 SETTING SW2 SETTING BAUD ODE __ 19200 GSIOC SLAVE EXTERNAL CLOCK 19200 GSIOC SLAVE INTERNAL CLOCK GSIOC MASTER BREAK ACTIVE GSIOC MASTER RS BRES ES o o 0000 O o o OOOO O LASA AH HY 240 Roa Qzev S w N a Zz Cc O 207 LO CAUTION FOR CONTINUED PROTECTION AGAINST RISK OF FIRE REPLACE ONLY WITH THE SAME TYPE AND RATING OF FUSE 5 Mains fuse drawer BREAK PASSIVE R R Rear panel connectors and switches GSIOC O 1 LED driver output for camera BE eae A 3 EE 2 7 3 Not used EI O 4 RS 232 port for connection to PC 1 O MAINS RS 232 O 6 Mains power switch 190 120 20 T i ea 7 Mains power socket ee a Unit ID select CE Q
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