User Manual - Spectra Services
Contents
1. 31 APPENDIX F DRIERITE MATERIAL SAFETY DATA SHEET 33 APPENDIX G LIMITED WARRANTY 35 APPENDIX H DISCLAIMER 37 INDEX u fk e ek kk Ba l i a Ka besi a ke ak Sak 39 TABLE OF FIGURES Figure 3 1 Components of the XenoWorks Digital MicroinjJector 3 Figure 3 2 Controller Compressor features front view essere tenens 5 Figure 3 3 Controller Compressor features rear view 5 Figure 3 4 Remote User Interface 7 Figure 4 1 Connecting the micropipette holder to the pressure tubing 9 Figure 4 2 Attaching the pressure fitting to other end of pressure tubing 10 Figure 7 1 Connecting the micropipette holder to the pressure tubing 22 TABLE OF TABLES 2 1 Mains fuse type and ratings iii euet ness breue dass daal 1 XENOWORKS DIGITAL MICROINJECTOR OPERATION MANUAL REV 4 13 20100914 Table 3 1 Pressure Vacuum range settings ii it 6 Table 5 1 Continuous mode ac2. 9 28 81 Micropipette holder diameter 31 mounting screws 29 O Operating Environment 81 OPERATING INSTRUCTIONS 11 packaging ee 29 Packing Taste see een 3 Piezo impact drive compatibility 81 Power Cord 3 precautions w aq 2 2 2 Pressure channels 81 Pressure Conversion 25 Pressure Pulse Width Display 8 Pressure Pulse Width Key 8 PRODUCT DESCRIPTION 3 Pulse Mode 8 14 Pulse Continuous Injection Mode Key 7 Regulatoria iu 31 remote user interface 3 4 5 safety warning 1 Shipping 4 eae n 9 29 shipping 29 shipping screws enne 29 Spare Parts hited tte dettes 23 Specifications 81 XENOWORKS DIGITAL MICROINJECTOR OPERATION MANUAL REV 4 13 20100914 40 Storage Environment 81 T technical specifications electrical intem 32 mains f se uite dese
3. 6 3 4 3 Range Setting for Transfer and Compensation Pressures 6 un ns Dy am OOO A 7 9 2 3 1010 DIA uod ipao a EA md empate d uy a Ad m ey ead 7 2 46 Hold DISDIAY us ott stretto 7 3 4 7 Pulse Continuous Injection Mode Key 7 3 48 Press Ur WISE Width Dial ee dash iden e a a b edd 8 3 4 9 Pressure Pulse Width Display U 8 3 4 10 Pressure Pulse Width Key wiccccccccccssessessevsescessessnsssssssesessessssesssssessssessessessessssssssesensessessessesseses 8 Clear Key nee ER 8 24 72 InJeeti a ans dee a ea 9 9 4 13 Transfer Dial cto OLIM 9 32 14 Transfer e ang asna 8 4 INSTALLATION Pcr p GG X 9 5 OPERATING INSTRUCTIONS case el 11 5 1 THE HOLD FUNCTION u ite A 11 5 2 THE TRANSFER FUNCTION ccccccssscccssssecssscssscecsssceessssesssscssseecsseeeessssesseecseeecseeeesscsessecsseeeessnees 12 5 2 1 Transfer Channel and Settin s un uuu 12 9 2 3 Performance AOVIGO a n An S T y aa Y 15 Did DHE INJECT FUNCTION 5 6 reed dae AI 14 5 3 1 Setting up the
4. 32 mains voltage 32 power consumption 32 power 32 technical support l a sus 1 Transfer Display u erint ctas ees 8 Transfer Function 11 12 18 Transfer pressure 81 transporting nia 29 Transporting the Microinjector 29 TROUBLESHOOTITING 17 Tubing etse 3 31 U user interface module 6 9 17 W Warranty n iocis eiie 35 a 32 XENOWORKS DIGITAL MICROINJECTOR OPERATION MANUAL REV 4 13 20100914
5. The XenoWorks Digital Microinjector Models BRE110 and BRE220 should only be used in a laboratory environment for use on animal tissues It is not intended for use nor should be used in human experimentation or applied to humans in any way This is not a medical device Do not open or attempt to repair the instrument without expressed and explicit instructions from Sutter Instrument Company Extreme heat and high voltages are present and could cause injury Do not allow unauthorized and or untrained operatives to use this device Any misuse will be the sole responsibility of the user owner and Sutter Instruments assumes no implied or inferred liability for direct or consequential damages from this instrument if it is operated or used in any way other than for which it is designed XENOWORKS DIGITAL MICROINJECTOR OPERATION MANUAL REV 4 13 20100914 39 INDEX A I allen wrench ua Hanne 29 C Cleaning x och awd o eos 32 Clear function eiecit aa irit ie 31 Clear Key tette eet ret tede es 8 COMPTESSOT een sense 10 compressor module 9 10 11 12 13 17 19 29 Continuous Mode 8 14 control DoX i A oot icis tire eee aee 3 controller compressor 4 controller compressor 29 Controller Compressor 3 Controls msn a muras 4 6 7 31 D Dime
6. order for more than five part BR DT minutes when the 2 The pressure fitting has not Push the pressure fitting into its port until instrument 18 been fully engaged in the a click is felt If no tubing is fitted to a but not being used pressure port on the front ofthe port check that the port is closed by compressor module pressing the release tab on top of the port 3 There is a loose pressure pressure fittings should be finger tight fitting no tools are necessary to tighten pressure fittings 4 There is no micropipette in the Insert a micropipette into the holder If no holder micropipettes are being used switch the Hold pressure off and turn the Transfer pressure down to zero If after performing the above checks the compressor motor continues to run for longer than five minutes shut the instrument down and contact your Sutter distributor The compressor is The shipping plate and screws on Turn off the compressor Place the cabinet loud when the right side panel have not been on its left side front side up and remove activated removed the hex head screws and shipping plate XENOWORKS DIGITAL MICROINJECTOR OPERATION MANUAL REV 4 13 20100914 20 Problem Cause Solution NOTE To replace the shipping plate before storage or shipping place the cabinet on its side again so that the receiving holes inside on the compressor line up with the external holes ERR appears on all B displa
7. Plastic tip of the pipette holder value as a guide only Make sure that the tubing connector is plugged in all the way into the port Carefully tighten the pipette holder s plastic tip is not tightened down XENOWORKS DIGITAL MICROINJECTOR OPERATION MANUAL REV 4 13 20100914 19 6 4 Inject Function Problem Cause Solution There is no 1 The microscope optics are Switch to suitable optics if available material flowing unable to resolve the stream Brightfield illumination will be unlikely to from the tip ofthe resolve the stream unless the injected micropipette even solution is colored Phase contrast will be when positive more effective and Nomarski and pressure is applied Hofmann better still A constant stream 80 psi 5566 hPa can also be detected by moving the on display micropipette tip close to a freely floating cell and trying to blow it away 2 The micropipette is blocked Press the Clear Key a number of times holding the key down longer each time until the blockage is cleared If this does not clear the blockage the pipette should be replaced CAUTION Some residual pressure may remain in the tubing Take great care to point the micropipette in a safe direction when removing it from the holder 6 5 General Problem Cause Solution Compressor motor 1 There is a leak in one of the Check each pressure line for kinks or runs continuously pressure lines holes Replace lines if necessary
8. properly engaged in the Hold Channel port the left hand port on the front of the compressor module and that all fittings are finger tight 2 Fit the holding micropipette into the micropipette holder by loosening the clear plastic chuck turn gently inserting the micropipette and tightening the chuck until the micropipette cannot be moved in or out of the holder CAUTION Do not over tighten the chuck doing so could crush the O rings and prevent them from making a airtight seal around the base of the micropipette Take care not to force the micropipette into a holder If the micropipette will not easily slide in it is because either the chuck is locked down or there is an obstruction such as broken glass in the holder Care should be taken to ensure the back end of the micropipette is clean and free of cracks or breakages to avoid damage to the O rings Despite these precautions the O rings will eventually wear and they should be inspected from time to time and replaced when necessary 3 Once the micropipette is fitted into the holder the holder can be locked into the micropipette holder clamp on the micromanipulator drive module and the tip of the micropipette lowered into the injection chamber Caution Take care not to let the holder snap into the spring loaded clamp since this may cause damage to the micropipette tip Turn the Hold Port on and dial the hold pressure up to 20 30 This will prevent oil or media from drawing into
9. 2 each including 2 5 m 8 ft clear ETFE tubing pressure fitting 5 spare ferrules Power cord Instruction Manual Controller Compressor Q Remote User Interface Pipette Holder Remote Interface 2 Cable Power Cord Foot Switch Manual Figure 3 1 Components of the XenoWorks Digital Microinjector XENOWORKS DIGITAL MICROINJECTOR OPERATION MANUAL REV 4 13 20100914 3 2 Instrument Design The XenoWorks Digital Microinjector is a pneumatic pressure control system for single cell microinjection and manipulation Two independently controlled pressure channels are available Hold and Inject Transfer The Hold Channel is designed to create negative and positive pressure sufficient to gently hold and release a single cell usually mammalian oocyte zygote or early embryo including fully hatched blastocyst stage embryos against a hollow fire polished glass micropipette Ascidian amphibian and insect embryos can be immobilized in a similar fashion depending upon their physical characteristics and the type of micropipette used The Inject Transfer Channel provides negative and positive pressures required to aspirate single cells such as embryonic stem cells into an appropriately fashioned micropipette in preparation for transfer to another location such as the blastocyst of an early mammalian embryo The transfer of cells is achieved by reversing the pressure in the tra
10. power cord To remove the canister from the device pull off the output left air tube from the white plastic connector on the canister 2 The two black plastic hold downs that secure the canister to the rear panel can be released by forcing one half of the connector out of the other half at the point where they meet Remove the canister and unscrew the plastic end cap being careful not to lose the black rubber sealing ring that forms the airtight seal within the cap 3 With the cap off remove the spring the perforated aluminum plate and the first filter and second filter The far aluminum plate can be kept in place Pour the Drierite out onto a glass or metal tray spreading it evenly one granule deep and heat it for one hour at 200 C Pre dry both filters at 100 C for 30 minutes just prior to assembly 4 Before refilling the canister the Drierite granules should be cooled in an airtight container After cooling replace the Drierite granules keeper and filters into the canister in the order they were removed 5 First install the filter against the keeper that was left in the canister 6 Ifthis keeper was removed slide it in first ensuring that it lays flat against the plastic shoulders at the far end of the canister With the far keeper and filter in place pour in the new or regenerated Drierite 7 Next insert the second filter followed by the keeper and the spring 8 Check that the rubber seal is in the proper position and i
11. that the instrument fails to power up when it is switched on the line power fuses should be checked to determine whether they have blown The fuses are located in the fuse holder in the power entry module on the rear of the control module To replace a fuse first unplug the power cord from the power entry module revealing the fuse holder below Remove the fuse holder Note that both fuses in the fuse holder are used simultaneously Either one or both may have blown Discard the blown fuse s Insert appropriately rated replacement fuses see below Replace the fuse holder in the power entry module and reconnect the power cord Replacement Fuse Ratings Table C 1 Mains fuse type and ratings Mains Fuse Voltage Type Time Delay 5mm x 20mm glass tube Setting use Rating Manufacturer Examples 110 VAC 110 4A 250V Bussmann GMC 4A GMC 4 R RoHS GDC 4A or 100 120 S506 4A RoHS VAC Littelfuse 239 004 239 004 P RoHS 218 004 or 218 004 P RoHS 220 VAC 220 2A 250V Bussmann GDC 2A or 5506 2 RoHS 200 240 Littelfuse 239 002 or 239 002 P RoHS VAC XENOWORKS DIGITAL MICROINJECTOR OPERATION MANUAL REV 4 13 20100914 29 APPENDIX D TRANSPORTING THE MICROINJECTOR When transporting use the original packaging and the supplied shipping screws and shipping plate Ifthe packaging and shipping screws have been misplaced contact your local Sutter distributor Install the four shipping sc
12. the pipette 4 When the micropipette is lowered into the medium a small quantity of medium will be aspirated by capillary action into the tip It is unnecessary to aspirate any more medium than this 5 With the Hold Pressure Dial set at approximately zero switch the Hold Function on The holding channel is now ready to use XENOWORKS DIGITAL MICROINJECTOR OPERATION MANUAL REV 4 13 20100914 12 6 Under certain conditions such as a broken micropipette it can become possible to aspirate fluid from the injection chamber through the tubing and into the compressor module through the front port Particular care should be taken not to allow this to occur To safeguard against this event a fluid sensor has been installed inside the controller If fluid is drawn into the controller the fluid sensor will be activated and a compensating positive pressure will turn onto expel the fluid The positive pressure will stay on continuously even if you power off and on the injector To avoid getting aspirating fluid into the controller a Making sure that the micropipettes are removed from the injection chamber when it is left unattended by for example using the micromanipulator s Home Function b Switch off the Hold Pressure Port and turn the Transfer pressure to zero when the device is to be left unattended If fluid has been inadvertently aspirated into the Hold Port tubing it can be purged by connecting the Hold Port to th
13. 0 A Power 280 VA For manufacturer examples refer to Table C 1 Mains fuse type and ratings XENOWORKS DIGITAL MICROINJECTOR OPERATION MANUAL REV 4 13 20100914 33 APPENDIX F DRIERITE MATERIAL SAFETY DATA SHEET IDENTITY INDICATING DRIERITE DATE PREPARED 1 3 96 DESCRIPTION 1 16 TO 1 4 BLUE GRANULES SECTION I MANUFACTURER S NAME W A HAMMOND DRIERITE CO LTD ADDRESS 0 BOX 460 138 DAYTON AVE XENIA OH 45385 EMERGENCY PHONE NUMBER 513 376 2927 INFORMATION PHONE NUMBER 513 376 2927 SECTION II INGREDIENTS CHEMICAL IDENTITY 5 PEL ACGIH TLV UNITS C A S CALCIUM SULFATE 97 15 10 mg M 7778 18 9 COBALT CHLORIDE 3 0 05 0 05 mg M 7646 79 9 AS COBALT METAL HAZARDOUS MATERIALS IDENTIFICATION SYSTEM HMIS HEALTH FLAMMABILITY REACTIVITY PROTECTIVE EQUIPMENT 1 0 1 SECTION III PHYSICAL CHEMICAL CHARACTERISTICS SPECIFIC GRAVITY H 0 1 1 87 SOLUBILITY IN WATER 0 25 GRAMS PER LITER MELTING POINT 1450 C DECOMPOSES APPEARANCE BLUE GRANULES NO ODOR SECTION IV FIRE AND EXPLOSION HAZARD DATA FLASH POINT NONE EXTINGUISHING MEDIA NOT COMBUSTIBLE SPECIAL FIREFIGHTING PROCEDURES NONE UNUSUAL FIRE AND EXPLOSION HAZARDS NONE SECTION V REACTIVITY DATA STABILITY STABLE INCOMPATIBILITY MATERIALS TO AVOID STRONG ACIDS HAZARDOUS DECOMPOSITION BYPRODUCTS CI 318 C SO 1450 C
14. 00914 APPENDIX A SPARE PARTS BR MH BR DT 870700 Micropipette holder includes micropipette holder body 9 O rings Digital tubing kit includes 2 x 2 m ETFE tubing pressure fitting 6 ferrules Drierite canister XENOWORKS DIGITAL MICROINJECTOR OPERATION MANUAL REV 4 13 20100914 23 25 APPENDIX B PRESSURE CONVERSION XenoWorks Digital Microinjector displays pressure in hecta Pascals hPa The following conversion factors should be used for other pressure units 1 hPa 100 Pascals Pa 1 Pascal 0 01 hPa 1 hPa 1 millibars 1 millibar 1 hPa 1 hPa 001 bars 1 bar 1000 hPa 1 hPa 0 0145 psi 1 psi 68 9655 hPa Table B 1 Pressure Vacuum range settings Range Scale Pressure Applications Setting Range 1 Unit Transfer Dial approx hPa hPa psi 1 Low 2 5to 2 5 172 to 1 0 015 Standard setting for ICSI 172 NT ES Cell Pronuclear Injection and Adherent Cell microinjection 2 Medium 5to 5 345 to 2 0 03 Used when higher 345 compensation pressures are needed for smaller volumes 3 Medium 10 to 7 689 to 4 back loaded into injection High 483 needle 4 High 15 to 7 1034 to 0 12 Used for font loading pipette 483 with very small volumes femtoliters and to create higher compensation pressures for these small volumes XENOWORKS DIGITAL MICROINJECTOR OPERATION MANUAL REV 4 13 20100914 27 APPENDIX FUSE REPLACEMENT In the event
15. Activate the holding pressure with the Hold on off switch green LED will light Connect the tubing to the left hand holding port There will be a positive click when the fitting is engaged Check the tubing for kinks holes and other damage and replace if necessary order part BR DT Tighten all fittings Replace ferrules Slowly rotate the Hold control clockwise until the air medium meniscus is as close to the tip as possible Note that it is possible to create an effective holding pressure and still show a positive value in the Hold LED display Carefully tighten the pipette holder s plastic tip Carefully tighten the rod holder s black parts located behind the plastic tip XENOWORKS DIGITAL MICROINJECTOR OPERATION MANUAL REV 4 13 20100914 18 6 3 Transfer Function Problem The Transfer Cause Solution Connect the tubing to the right hand pressure control seems unresponsive 1 The tubing is not connected to the transfer pressure port 2 There is a leak in the tubing or the fittings are loose 3 The air medium interface is too far inside the micropipette tip 4 The plastic tip of the pipette holder is not tightened down 5 The black parts on the rod holder behind the plastic tip are loose inject transfer port There will be a positive click when the fitting is engaged Check the tubing for kinks holes and other damage and replace if necessary order part BR DT
16. Before operating the instrument check that the instrument s voltage rating corresponds to the supply voltage The voltage rating can be found on the power entry module on the rear of the instrument Before making electrical connections ensure that the instrument is switched off AN Replace fuse only with the same type and rating as indicated in the following table Table 2 1 Mains fuse type and ratings Mains Mains Fuse Power Voltage Type Time Delay 5mm x 20mm glass tube Source Setting Fuse Rating Manufacturer Examples 110 VAC 110 4A 250V Bussmann GMC 4A GMC 4 R RoHS GDC 4A or 100 120 S506 4A RoHS VAC Littelfuse 239 004 239 004 P RoHS 218 004 or 218 004 P RoHS 220 VAC 220 2A 250V Bussmann GDC 2A or S506 2A RoHS 200 240 Littelfuse 239 002 or 239 002 P RoHS VAC A spare fuse is provided with fuse holder located in the power input module Please refer to Appendix C Fuse Replacement for instructions on how to change the fuse prevent fire or shock hazard not expose the unit to rain or moisture To avoid electrical shock and exposure to hazardous electrical voltages not disassemble the unit Refer servicing to qualified personnel Always use the grounded power supply cord provided to connect the unit to grounded outlet 3 prong This is required to protect you from injury in the event that an electrical hazard develops To comply with FDA
17. G CONDITIONS WASTE DISPOSAL METHOD THIS MATERIA SOLID WASTE IN AN APPROVED LAND FIL STATE AND LOCAL REGULATIONS PRECAUTIONS TO BE TAKEN IN HANDLING AND STORING KEEP CONTAINER CLOSED STORE IN A COOL DRY PLAC AVOID GENERATING DUST F E WASTE x R FOR DISPOSAL AVOID CAN BE DIS OR BY OTH POSI R P ED OF A ROCEDUR 5 AN INERT ES ACCEPTABL EX h h ER F ED ERAL F F Di SECTION VIII CONTROL MEASURES HA APPROVED FOR DUST RESPIRATORY PROTECTION NIOSH OS E IENTS VENTILATION TO MEET TLV REQUIR EYES SAFETY GLASSES OR GOGGLES OTHER PROTECTIVE EQUIPMENT GLOVI MAY BE DESIRAB IN SPECIFIC WO T T F FS OR K SITUATIONS CLOTHING NOT USUALLY N EC ESSARY F Nun SECTION IX REFERENCES 0 5 DEPARTMENT OF LABOR OSHA FORM APPROVED 0 1218 0072 OSHA HAZARD COMMUNICATION STANDARD 29 CFR 1910 1200 U S GYPSUM CO Although the information and recommendation set forth herein are presented in good faith and believed to be correct as of the date hereof the W A Hammond DRIERITE Co makes no representation as to the completeness or accuracy thereof Information is su
18. HAZARDOUS POLYMERIZATION WILL NOT OCCUR SECTION VI HEALTH HAZARD DATA EYES PARTICLES MAY CAUSE IRRITATION SKIN THIS MATERIAL IS NOT TOXIC MAY DRY OR IRRITATE SKIN INHALATION MAY CAUSE AN IRRITATION OF RESPIRATORY ORGANS OF SENSITIVE RESULTING IN THE OBSTRUCTION OF AIRWAYS WITH SHORTNESS OF BREATH INGESTION MAY CAUSE VOMITING DIARRHEA AND SENSATION OF WARMTH SIGNS AND SYMPTOMS OF OVER EXPOSURE EYE NOSE THROAT OR RESPIRATORY IRRITATION FU ERSONS XENOWORKS DIGITAL MICROINJECTOR OPERATION MANUAL REV 4 13 20100914 CARCINOGENICITY OF INGREDIENTS MATERIAL CALCIUM SULFATI NOT LIST COBALT CHLORIDI YES NO COBALT amp COBAI POUNDS ARE CLASSIFIED AS GROUP 28 MEDICAL CONDITIONS GENERALLY AGGRAVATED BY EXPOSURE PRE EXISTING UPPER RESPIRATORY AND LUNG DISEAS BRONCHITIS EMPHYSEMA amp ASTHMA EMERGENCY AND FIRST AID PROCEDURES EYES FLUSH WITH WAT DUST INHALATION KIN WASH WITH WAT S INGESTION NTP NOT LIST OSHA NOT LIST NO F Di D SUCH 5 NOT LIMIT ED TO F TO FR ESH AIR F NONE SECTION VII SPILL OR LEAK PROCEDURES STEPS TO BE TAKEN IN CASE MATERIAL IS RELEASED OR SPILLED SWEEP OR VACUUM MATERIAL INTO APPROPRIAT CONTAIN DUSTIN
19. High Pressure Injection Channel and Micropipette 14 HARE AOI TINT OTS MIO m ne een 14 ODD e E VI OD Ca pae ti dada dede B A AT 14 6 TROUBLESHOOTING 17 6 1 INSTRUMENT POWER asien tit eee eret ee eot alee ceti eae asa uya as 17 6 2 HOLD FUNETION i e A cete eee decet eu Fee ee A ce M RU CORDE 17 6 3 TRANSFER FUNCTION S RP Su Qiu 18 XENOWORKS DIGITAL MICROINJECTOR OPERATION MANUAL REV 4 13 20100914 IV 6 4 INJECTEUNGCTION na eh e ee est aa aW u tte S ec REUS 19 i786 oho PUER ET 19 TMATN TENAN Cn A egal Lcd epu ee 21 7 1 REGENERATION OF THE DRIERITE 21 1 2 FITTING NEWOSRINGS aha aa nhu t a amaga h 22 APPENDIX SPARE PARTS 23 APPENDIX PRESSURE CONVERSION 25 APPENDIX C FUSE REPLACEMENTA A 27 REPLACEMENT FUSE RATINGS 27 APPENDIX D TRANSPORTING THE MICROINJECTOR 29 APPENDIX E SPECIFICATIONS
20. OPERATION MANUAL 4 13 20100914 XENOWORKS DIGITAL MICROINJECTOR INSTRUMENT XENOWORKS DIGITAL MICROINJECTOR OPERATION MANUAL REV 4 13 20100914 SUTTER INSTRUMENT COMPANY ONE DIGITAL DRIVE NOVATO CA 94949 VOICE 415 883 0128 WEB WWW SUTTER COM FAX 415 883 0572 EMAIL INFO SUTTER COM Copyright 2009 Sutter Instrument Company All Rights Reserved XenoWorks is a trademark of Sutter Instrument Company XENOWORKS DIGITAL MICROINJECTOR OPERATION MANUAL REV 4 13 20100914 TABLE OF CONTENTS L GENERALINFORMATJON o atop hM ua d p eid d e bam UD 1 JE ABOUT THIS MANUAL a si ie a n Q ete nieht 1 1 2 TECHNICAL SUPPORT ii a ii ba A Pede dit ei eere A 1 2 SAFETY WARNINGS AND PRECAUTIONS 1 PAE EEECTRICAE Bea ee c ay M in 1 2 2 PRECAUTIONS Yale a e e 2 9 PRODUCT DESCRIPTION 3 3 1 PACKING LIST ia aaa aa be etes ete e a n e TREO SR 3 3 2 INSTRUMENT DESIGN S u a etuer iste en aa e due eoe sa 4 3 3 IDENTIFICATION OF INSTRUMENT COMPONENTS AND CONTROLS 4 QIS LA AA AAA 5 3 4 GONTROLSAND FEATURES 000 nil a nutu S air Rain ras 6 Port uu ul nanan 6 3 4 2 Inject Transter Port suyu wn 3222 Qa Dn n
21. Tighten all fittings Replace ferrules Slowly rotate the Transfer control clockwise until the air medium meniscus is as close to the tip as possible Carefully tighten the pipette holder s plastic tip Carefully tighten the rod holder s black parts located behind the plastic tip Note that the Transfer LED display is a guide only and that it is possible to have a fully equalized micropipette no net in or out flow of material and still show a positive or a negative i e non zero value in the display If after performing the above checks the device is still unresponsive please contact your Sutter Instrument distributor for assistance The Transfer 1 Factors such as viscosity of This is normal Use the display pressure display shows a positive or negative pressure yet the cells are stationary inside micropipette medium and capillary surface tension forces create a negative pressure in the micropipette It may take slight POSITIVE pressure from the microinjector to balance this action and even with a net negative pressure at the tip the pressure applied to the tool may be slightly positive If the Transfer pressure can be dialed up or down with no effect on the aspirated cells there may be a problem with the pressure hose connection see The Transfer pressure control seems unresponsive above 2 Tubing connector not plugged in all the way into the port on the front panel of the digital injector 3
22. and CE electromagnetic immunity and interference standards and to reduce the electromagnetic coupling between this and other equipment in your lab always use the types and lengths of interconnect cables provided with the unit for the interconnection of the remote user interface and foot switch XENOWORKS DIGITAL MICROINJECTOR OPERATION MANUAL REV 4 13 20100914 2 2 Precautions Failure to comply with any of the following precautions may damage this device Please read this manual carefully before operating the instrument Use this instrument only for microinjection purposes in conjunction with the procedures and guidelines in this manual Do not operate if there is any obvious damage to any part of the instrument Do not operate this instrument near flammable materials Do not obstruct the side vents or cooling fan intakes As with all microinjection devices sharp micropipettes can fly out of their holder unexpectedly Always take precautions to prevent this from happening Never loosen the micropipette holder chuck when the tubing is pressurized and never point micropipettes at yourself or others Always wear safety glasses when using sharp glass micropipettes with pressure microinjectors This instrument is designed for use with capillary glass micropipettes with an outer diameter of 1mm For glass with outer diameters of 1 2 and 1 5 mm please contact Sutter Instrument to order an additional holder Adapters for other capill
23. ary diameters are available upon request Use this instrument only for microinjection purposes in conjunction with the procedures and guidelines in this manual Please retain the original packaging for future transport of the instrument Some applications such as piezo impact microinjection call for the use of mercury in the micropipette tip The use of any hazardous materials with any XenoWorks instrument is not recommended and undertaken at the users own risk Always transport the instrument in its original packaging and install shipping plate screws before shipping This instrument contains no user serviceable components do not open the instrument casing This instrument should be serviced and repaired only by Sutter Instrument or an authorized Sutter Instrument servicing agent This device is intended only for research purposes Sutter Instrument reserves the right to change specifications without prior notice XENOWORKS DIGITAL MICROINJECTOR OPERATION MANUAL REV 4 13 20100914 3 PRODUCT DESCRIPTION 3 1 Packing List XenoWorks Digital Microinjector is shipped with the following components If any items are missing or damaged contact Sutter Instrument immediately o Digital Microinjector cabinet controller compressor Digital Microinjector control box remote user interface Cable to connect remote user interface with controller Foot switch Micropipette holder 2 Tubing kits
24. compatible with a number of different micromanipulators the setup procedure used with those micromanipulators may vary slightly 1 Unpack the XenoWorks Digital Microinjector 2 Remove the 4 shipping screws and plate from the right side of the instrument Retain plate amp screws for possible future transport 3 Connect the microinjector compressor and user interface modules using the cable provided 4 Place the compressor module on a flat solid surface away from the microscope on a different surface from microscope s is strongly suggested 5 Ensure that the compressor is located close enough to the microscope that the pressure lines will reach without placing any strain on the micropipette holders or micromanipulators The user interface module can be located conveniently next to the microscope 6 Remove the micropipette holders from their cases and connect them to the free end of the pressure tubing as shown below O RINGS 2 Figure 4 1 Connecting the micropipette holder to the pressure tubing XENOWORKS DIGITAL MICROINJECTOR OPERATION MANUAL REV 4 13 20100914 10 9 Attach a pressure fitting to the other end of the pressure tubing as shown in below Do not use tools to tighten pressure fittings since damage may result Figure 4 2 Attaching the pressure fitting to other end of pressure tubing 10 Connect each of the pressure lines to the appropriate ports of the compressor module You will hea
25. cropipette holder by sliding it up or down the clamp so that the tip of the micropipette projects into the optical axis of the microscope For details on aligning micropipettes please refer to the instruction manual of the micromanipulator If using a micropipette with a bend rotate the holder in the clamp until the bend section is parallel to the bottom of the injecting chamber Turn the Transfer Pressure Dial 80 and lower the micropipette into the injection medium XENOWORKS DIGITAL MICROINJECTOR OPERATION MANUAL REV 4 13 20100914 13 5 A small amount of medium will be drawn into the micropipette by capillary action Wait approximately 30 seconds for the pressure to equalize and then slowly turn the Transfer Dial clockwise until the meniscus between air and medium is less than approximately 2 mm from the micropipette tip The Transfer Dial should read between 060 to 090 at this point 6 The micropipette can now be loaded with for example embryonic stem cells by placing the tip close to the material to be aspirated and slowly turning the Transfer Dial counter clockwise to lower settings e g between 30 70 7 Expelling cells from the micropipette is achieved by gently turning the dial clockwise in much the same way as with a standard micrometer microinjector 5 2 3 Performance Advice The XenoWorks Digital Microinjector uses precision regulated air pressure to hold inject and manipulate cells and other tissue sam
26. e Inject Transfer Port Then pressing the Clear key Fluid in the Hold Port tubing can also be purged by turning the Hold Dial to maximum positive pressure until the fluid is drained Never leave the unit unattended when there is fluid in the tubing If this happens contact Sutter Instrument for tech support and instructions on how to resolve this matter 5 2 The Transfer Function The Transfer Function works in a similar manner to the Hold Function but is always active there is no on off control Pressure ranges from 175 hPa to 175 hPa 7 hPa Range 1 and is applied to the pressure port used to control the pressure in the transfer inject micropipette A typical application for which this function might be used includes the transfer of embryonic stem cells into a blastocyst Caution Do not shut down and restart the device with a micropipette immersed in the dish since during the device s initialization process a small amount of pressure is sometimes released 5 2 1 Transfer Channel and Settings 1 3 As with the holding channel all fittings should be tight and fully engaged and the transfer micropipette inserted into the holder with care Remember not to tighten the chuck too far or the O rings will be crushed Insert the micropipette holder into clamp on the micromanipulator Take care not to let the holder snap into the spring loaded clamp since damage may occur to the micropipette tip Adjust the mi
27. endently controllable 5600 hPa 50 hPa 0 to 5600 hPa 50 hPa in 7 hPa increments 0 01 to 10 seconds In 0 01 sec increments Transient hand or foot switch 175 to 175 hPa 7 hPa in 7 hPa increments 350 to 350 hPa 7 hPa in 7 hPa increments Ramp from Inject value to maximum pressure 3x 7 segment LED Tactile keys rotary optical encoders 2mETFE 4 1 mm capillary glass other sizes available upon request Prime Tech Burleigh 0 70 C 32 158 F 0 95 non condensing 3 5 35 C 38 3 95 F 0 80 non condensing 80 31 C 87 8 F decreasing linearly to 67 35 C 95 EN61010 1 TUV CE EN61326 TUV CE XENOWORKS DIGITAL MICROINJECTOR OPERATION MANUAL REV 4 13 20100914 32 Cleaning Dimensions H x W x D Weight Controller compressor Remote user interface Electrical Mains voltage Power consumption Power cord Mains fuse rear of cabinet Input power BRE110 Input power BRE220 70 alcohol or e g UV Compressor module 407 x 440 x 150 mm 16 x 17 x 6 inches User interface module 164 x 123 70 mm 6 5 x 4 8 x 2 8 inches 1504 g 3 36 Ib 3 Ib 5 oz 42 g 0 093 Ib 1 48 2 100 240 V 50 60 H 440W 10A 250V with safety ground plug Time delay or time lag 5 x 20 mm glass tube 100 120 VAC x 1096 50 60 Hz Fuse rating 4 0 A Power 450 VA 220 240 VAC 10 50 60 Hz Fuse rating 2
28. er Compressor features rear view 8 3 1 Drierite Canister The XenoWorks Digital Microinjector incorporates an integral compressor which takes its air supply from the ambient air The canister of Drierite granules dries the air before it is compressed to ensure a minimum of condensation and a long working life for the compressor components The blue Drierite granules slowly turn pink as they absorb moisture from the input air Once all the granules have turned pink the Drierite should be desiccated or replaced described in the Maintenance chapter later in this manual The approximate life of the Drierite canister is one year under normal conditions XENOWORKS DIGITAL MICROINJECTOR OPERATION MANUAL REV 4 13 20100914 3 4 Controls and Features 3 4 1 Hold Port This port supplies a pressure range from 350 hPa to 350 hPa 7 hPa and is typically used to control the pressure in the holding micropipette The pressure can be varied using the Hold Dial on the remote user interface 3 4 2 Inject Transfer Port This port supplies four possible transfer compensation ranges Range setting 1 is the default setting providing from 175 hPa to 175 hPa 7hPa and is typically used to control the pressure in the transfer micropipette during embryonic stem cell transfer experiments and to create compensation pressure for high pressure injection of solutions The compensation pressure can be varied using the Transfer Dial on the user int
29. erface module The pressure pulse duration for pronuclear and adherent cell injection can be set with the Pressure Pulse Width dial 3 4 3 Range Setting for Transfer and Compensation Pressures This rotary switch is used to set the compensation pressure range on the Inject Transfer port The following table shows the compensation pressure range for each of the four settings Table 3 1 Pressure Vacuum range settings Range Scale Pressure Applications Setting Range 1 Unit Transfer Dial approx 1 Low 2 5to 2 5 172 to 1 0 015 Standard setting for ES Cell 172 Pronuclear Injection and Adherent Cell microinjection 2 Medium 5to 5 345 to 2 0 08 Used when higher 345 compensation pressures are needed for smaller volumes P Mediam back loaded into injection High 483 needle 4 High 15 to 7 1034 to 0 12 Used for front loading 483 pipette with very small volumes femtoliters and to create higher compensation pressures for these small volumes XENOWORKS DIGITAL MICROINJECTOR OPERATION MANUAL REV 4 13 20100914 PRESSUREIPULSE WIDTH DISPLAY PRESSUREIPULSE HOLD DISPLAY DIAL TRANSFER DISPLAY l DL HOLD PRESSURE TRANSFER PULSE WIDTH HOLD DIAL TRANSFER DIAL HOLD KEY INJECT KEY PRESSURE CONTINUOUS CLEAR KEY INJECTION MODE KEY PRESSURE PULSE WIDTH KEY Figure 3 4 Remote User Interface 3 4 4 Hold Key This key is used to toggle the Hold pressure on and of
30. f In the OFF position no pressure is applied to the Hold Port In the ON position the pressure in hPa shown in the Hold LED Display above the Hold Dial is applied to the Hold Port 3 4 5 Hold Dial Controls the pressure delivered to the Hold Port when the Hold Key is in the ON position The range is continuous from 350 hPa to 4350 hPa 7 hPa 3 4 6 Hold Display Displays the pressure delivered to the Hold Channel in hPa 3 4 7 Pulse Continuous Injection Mode Key This key toggles between the pulse mode and continuous mode used for high pressure injections The pulse pressure and duration are set with the Pressure Pulse Width Dial The continuous pressure is active as long as the Inject Key or footswitch is held down The mode selected and active is indicated with a green LED XENOWORKS DIGITAL MICROINJECTOR OPERATION MANUAL REV 4 13 20100914 3 4 8 Pressure Pulse Width Dial This dial is used to adjust the pressure and duration of injection Injection pressure can be adjusted from 0 to 5600 50 hPa 70 hPa increments Pulse duration can be adjusted from 0 01 to 10 seconds in 0 01 second increments 3 4 9 Pressure Pulse Width Display This display indicates either the injection pressure in hPa or the injection duration in seconds when in Pulse Mode Toggling between these two values is accomplished with the Pressure Pulse Width Key 3 4 10 Pressure Pulse Width Key This key is used to toggle the Pressure Pu
31. f you wish apply a thin layer of vacuum grease on to the rubber seal to ensure a tight fit There is no need to over tighten the cap but a good seal is needed to prevent air leaks 9 Reinstall the canister on the injector with the air tube connections to the top 10 Slide the black plastic hold downs inside one another and squeeze them together to firmly hold the canister in place 11 At this point re connect the air output tube the one which enters into the rear panel of the injector CAUTION Failure to maintain the Drierite may result in impaired instrument performance and can damage the instrument XENOWORKS DIGITAL MICROINJECTOR OPERATION MANUAL REV 4 13 20100914 22 7 2 Fitting New O rings Routine use will create wear on the three black O rings located in the tip of the micropipette holder chuck Occasionally the O rings will need to be replaced To do this gently remove the old O rings with an appropriate tool a bent paper clip for example Discard the old O rings and insert three new ones taking care to ensure that they are flat against the back of the chuck The new O rings can be pushed down inside the chuck by gently screwing the chuck onto the knurled black aluminum pressure fitting Take care not to damage the chuck during this procedure O RINGS 2 Figure 7 1 Connecting the micropipette holder to the pressure tubing XENOWORKS DIGITAL MICROINJECTOR OPERATION MANUAL REV 4 13 201
32. lay The display is showing only the pressure supplied to the micropipette even though the pressure effects observed at the micropipette tip are a combination of pressure supplied by the injector and capillary forces CAUTION Under certain conditions such as a broken micropipette it can become possible to aspirate fluid from the injection chamber through the tubing and into the compressor module through the front port Partieular care should be taken not to allow this to happen since the compressor module could be damaged Safeguard against such damage by 1 Making sure that the micropipettes are removed from the injection chamber when it is left unattended by using the micromanipulator s Home Function for example 2 Switch off the Hold Pressure Port and turn the Transfer pressure to zero when the device is to be left unattended XENOWORKS DIGITAL MICROINJECTOR OPERATION MANUAL REV 4 13 20100914 14 Fluid that has been inadvertently aspirated into the Inject Transfer tubing can be purged by pressing the Clear Key ensure there is no micropipette in the holder before doing this Never leave the unit unattended when there is fluid the tubing 5 3 The Inject Function 5 3 1 Setting up the High Pressure Injection Channel and Micropipette If the digital microinjector is being used for the high pressure injection of solutions such as DNA RNA proteins etc into cells through narrow microinjection micropipettes tip size
33. lse Width Dial Display between Pressure and Pulse Width 3 4 11 Clear Key Applies maximum pressure to the right hand Inject Transfer channel and is used to clear blockages of the micropipette When the key is pressed and held down the pressure applied increases from zero to maximum pressure 80 psi CAUTION THE CLEARING FUNCTION SHOULD NOT BE USED WITH MICROPIPETTES THAT HAVE TIP INNER DIAMETERS GREATER THAN APPROXIMATELY ONE 1 3 4 12 Inject Key This key applies the pressure set by the center Pressure Pulse Width Dial to the Inject Transfer pressure port When the instrument is in Pulse Mode the duration of the pressure will be for the time set by the Pulse Width Dial When the instrument is in Continuous Mode the pressure is applied for as long as the Inject Key or footswitch is held down 3 4 13 Transfer Dial This dial controls the pressure continuously supplied to the Inject Transfer Port Range Setting 1 standard setting provides 175 hPato 175 hPa 7 hPa of pressure 3 4 14 Transfer Display This display indicates the low pressure being constantly delivered to the Transfer Channel in hPa XENOWORKS DIGITAL MICROINJECTOR OPERATION MANUAL REV 4 13 20100914 4 INSTALLATION NOTE These instructions refer to the use of the XenoWorks Digital Microinjector with the XenoWorks Micromanipulator While the Digital Microinjector is supplied with micropipette holders that are
34. nsfer micropipette and gently injecting There are four range settings available for the Transfer Compensation channel allowing for the fine tuning of the pressure range for a given application In addition to these low positive and negative pressures the Inject Transfer Channel is capable of providing the high positive pressure required to inject solutions through very narrow typically less than one micron injection micropipettes during for example the injection of DNA solution into the pronucleus of a mammalian 2PN stage zygote Users should refer to later sections of this manual for more information on specific microinjection applications and to the application note Introduction to Microinjection available from Sutter upon request or for download from http www sutter com 3 3 Identification of Instrument Components and Controls The following figures illustrate the features on the front and rear of the controller compressor and the controls on the remote user interface XENOWORKS DIGITAL MICROINJECTOR OPERATION MANUAL REV 4 13 20100914 Spring release and push down on metal tab to remove tubing connector ON OFF SWITCH HOLD PORT RANGE CONTROL INDER UE INJECT TRANSFER FOR COMPENSATION LIGHT xi PORT PRESSURE Figure 3 2 Controller Compressor features front view POWER INPUT MODULE FOOT SWITCH CONNECTION REMOTE USER INTERFACE F CONNECTION DRIERITE CANISTER Figure 3 3 Controll
35. nsions 2d etr d tren Een aes 32 disclaimer Ai Ai tet tes 37 Display nu lets 7 8 31 Drierite nen lan lan 5 21 23 Drierite Canister 5 drierite material safety data sheet 33 E ETEEt bing 5 eet 3 23 F Features ten etc ertet teint aatete ees obs 6 Foot switch um E aa a 3 Fuse Replacement 27 fuses replacement MAINS m e este 1 G General Safety Information 1 H hecta Pascals eese 25 High pressure 14 31 Hold Channel tii iia 11 Hold Dial L REOR de 7 Hold Displays ee iil Rete egt tios 7 Hold Function 11 17 Hold Key ise oer SSS ae 7 Hold Port a sre Gaya uq ahus kaa 6 Hold pressure 31 Inject activation 31 Inject Function eese 14 19 Inject Transfer Port 20 6 Injection 31 Input power BRF110 32 Input power BRE220 32 INSTADDATION tte et 9 Instrument Design 4 M mains Uses ix sad d D UR TREE 1 Maximum pressutre 31 Micropipette compatibility 31 Micropipette holder
36. nuous mode actions and controls Action Control Used Adjust compensation pressure Transfer dial Select continuous injection Pulse Cont key to Cont mode Select high pressure Pressure Pulse Width key to adjustment Pressure Adjust high pressure value Pressure Pulse Width dial Inject Inject key or footswitch 5 3 3 Pulse Mode Pulse Mode provides high pressure injection by discrete pulses of a given pressure in hPa for a predetermined amount of time Pressure is applied for a predetermined duration no matter how long the Inject Key or footswitch is pressed This mode is useful for the injection of DNA into the pronucleus of amammalian zygote for example where successful injection XENOWORKS DIGITAL MICROINJECTOR OPERATION MANUAL REV 4 13 20100914 15 requires a high pressure and a slight inflating approximate doubling in size of the pronucleus can be seen Table 5 2 Pulse mode actions and controls Action Control Used Adjust compensation pressure Transfer dial Select pulse injection mode Pulse Cont key to Pulse Select high pressure Pressure Pulse Width key to adjustment Pressure Adjust high pressure value Pressure Pulse Width dial Select injection duration Pressure Pulse Width key to adjustment Pulse Width Adjust injection duration Pressure Pulse Width dial Inject Inject key or footswitch Once the base pressure injection pressure and if used injection duration have been set the sys
37. ples best control is attained when the interface meniscus between the air in the micropipette and the medium in the dish is as close as possible to the tip of the micropipette The optimum position of the meniscus will depend upon a number of factors including the type size and diameter of the micropipette the viscosity of the medium and the depth of the injecting chamber In general though the closer to the micropipette tip the meniscus can be brought the better the control over the pressure During the course of an experiment it may be necessary to ensure that the meniscus has not been allowed to be pulled too far up inside the micropipette Ifthe meniscus is too close to the micropipette tip control can become too sensitive and will not be smooth This is particularly so with embryonic stem cell transfer micropipettes If this is the case simply draw a little more medium into the micropipette by turning the Transfer Dial counter clockwise CAUTION Care should be taken not to apply too much positive pressure to the micropipette as with any other microinjection device it is possible to purge all the medium from the micropipette and stream bubbles into the injection chamber which may damage tissues and obscure the microscope opties NOTE The pressure reading on the LED display is a guide only it is possible to maintain embryonic stem cells stationary in the transfer micropipette tip yet still show a positive pressure in the disp
38. pplied upon the condi suitabili Co be re reliance expressed other nat informati tion that the person receiving same will make their own determination as to its F ty for their purpose prior to use In no event will the W A Hammond DRI sponsible for damages of any nature whatsoever resulting from the use of or upon information herein supplied No representations or warranties either or implied of merchantability fitness for a particular purpose of or any ure are made hereunder with respect to information or the product to which on refers XENOWORKS DIGITAL MICROINJECTOR OPERATION MANUAL REV 4 13 20100914 35 APPENDIX G LIMITED WARRANTY Sutter Instrument Company a division of Sutter Instrument Corporation limits the warranty on this instrument to repair or replacement of defective components for one year after the date of shipment provided the instrument has been operated in accordance with the instructions outlined in the instruction manual Abuse misuse or unauthorized repairs will void this warranty Limited warranty work will be performed only at the factory and the cost of shipment both ways is to be borne by the user The limited warranty is as stated above and no implied or inferred liability for direct or consequential damages is intended XENOWORKS DIGITAL MICROINJECTOR OPERATION MANUAL REV 4 13 20100914 37 APPENDIX H DISCLAIMER
39. r a click sound as the fittings engage in the port 11 Connect the power cord to the power input module on the rear ofthe compressor and a mains power outlet The XenoWorks Digital Microinjector is now set up and ready for use XENOWORKS DIGITAL MICROINJECTOR OPERATION MANUAL REV 4 13 20100914 11 5 OPERATING INSTRUCTIONS 5 1 The Hold Function If injecting adherent cells or if no Hold Function is required skip this section and move on to the Transfer Function The left hand rotary control and its associated LED display correspond to the pressure control of the left hand port on the front of the compressor module and of any tubing micropipette holder and micropipette attached to that port The pressure delivered by this port ranges from 350 hPa to 350 hPa 7hPa and is designed to allow a typical holding micropipette to hold amammalian oocyte blastocyst or zygote for example The Hold Function can be used a similar fashion to that of a traditional micrometer type air microinjector by dialing the pressure up or down to release or hold the cell in place A setting of 20 30 will advance meniscus to the top and reducing pressure to 10 will being to draw in the cell or egg Setting up the holding channel and micropipette 1 First ensure that the Hold Function is switched off the default position of the Hold Function after first powering up the unit then check that the tubing and fitting are
40. rews and the shipping plate to the right side of the compressor module using the following procedure 1 Align shipping plate holes with holes in the right side of the instrument 2 Mount the shipping plate using the four 10 32 socket head shipping screws provided by placing screws first through plate then through instrument side panel and finally engaging screws into interval motor bracket 3 Slowly advance the shipping screws in evenly to secure the compressor before storage or shipping 4 Tighten the 4 shipping screws using a 5 32 allen wrench 5 The controller compressor is now ready for transport and can be placed into the original foam packaging for shipping CAUTION Do not transport the compressor module without using the provided shipping screws and plate or instrument damage may result If screws plate misplaced and unavailable please contact Sutter Instrument to obtain the proper hardware XENOWORKS DIGITAL MICROINJECTOR OPERATION MANUAL REV 4 13 20100914 APPENDIX E SPECIFICATIONS 31 Pressure channels Maximum pressure High pressure Injection time Inject activation Transfer pressure Hold pressure Clear function Display Controls Tubing Micropipette holder diameter Micropipette compatibility Piezo impact drive compatibility Storage Environment Temperature Humidity Operating Environment Temperature Humidity Regulatory Safety EMC C 2 indep
41. s less than 1 micron the Transfer Function should be used as an adjustable positive pressure base pressure applied to the tip of the injection micropipette This ensures that there is a gentle constant flow of solution from the micropipette preventing dilution of the solution to be injected by the medium in the injection chamber The base pressure also minimizes to some degree blockage of the micropipette tip by foreign objects in the surrounding medium The wider the tip of the injection micropipette the less base pressure is required Asa rule start at 5 hPa and increase slowly until a gentle stream of material can be seen coming from the tip Under certain microscope optics this stream is easily visible as a Schlieren pattern resulting from the different densities of injected solution and surrounding medium Once the stream can be seen issuing from the micropipette tip reduce the base pressure until the stream almost disappears Two modes of high pressure injection are available to suit different users preferences 5 3 2 Continuous Mode High pressure of a predetermined value is applied for as long as the Inject Key or the footswitch is held down This mode is useful when injecting a field of cultured adherent cells where long durations and or high pressures will rupture and kill the cells The pressure can be set low and the duration short to deliver tiny discrete and reproducible volumes to each cell Table 5 1 Conti
42. tem is ready to begin injection Consult the application note Introduction to Microinjection for more information on microinjection techniques XENOWORKS DIGITAL MICROINJECTOR OPERATION MANUAL REV 4 13 20100914 6 TROUBLESHOOTING 17 6 1 Instrument Power Problem The instrument does not activate when the power is switched on 6 2 Hold Function Problem The holding pressure is insufficient to hold a cell or the hold control seems unresponsive Cause 1 The power cord is not connected to an appropriate supply 2 The protection fuse has blown 3 The user interface module is not properly connected Cause 1 The holding pressure is not activated 2 The tubing is not connected to the holding pressure port 3 There is a leak in the tubing or the pressure fittings are loose 4 The air medium interface is too far inside the micropipette tip 5 The plastic tip of the pipette holder is not tightened down 6 The black parts on the rod holder behind the plastic tip are loose Solution Connect the power cord to the mains supply Be sure that the power rating is matched to the requirements of the instrument Catalog number BRE110 requires 100 110v AC BRE220 requires 220 240v AC Replace the fuse and check that the power supply is appropriate to the requirements ofthe device Connect the user interface module to the compressor module using the cable provided Solution
43. tions and controls 14 Table 5 2 Pulse mode actions and controls er 15 Table B 1 Pressure Vacuum range settings 25 Table C 1 Mains fuse type and 27 XENOWORKS DIGITAL MICROINJECTOR OPERATION MANUAL REV 4 13 20100914 1 GENERAL INFORMATION 1 1 About This Manual The XenoWorks Digital Injector is a injection system comprised of three basic parts the controller compressor injection tubing and the user interface remote and foot switch In the next few pages you will find a product description to help you become acquainted with the parts followed by installation instructions and then detailed operating instructions Please take the time to read these instructions to assure the safe and proper use of this instrument 1 2 Technical Support Unlimited technical support is provided by Sutter Instrument Company at no charge to our customers Our technical support staff is available between the hours of 8 00 AM and 5 00 PM Pacific Time at 415 883 0128 You may also E mail your queries to info sutter com XENOWORKS DIGITAL MICROINJECTOR OPERATION MANUAL REV 4 13 20100914 2 SAFETY WARNINGS AND PRECAUTIONS Please read this manual carefully before operating the instrument 2 1 Electrical Use only a properly grounded power source and power cord both appropriately rated for use with this instrument
44. ys of the remote user interface 1 There is an electronic short on Contact Sutter Instrument the circuit board 2 There is a constant continuous Contact Sutter Instrument positive pressure and the compressor is always on It is possible that the solution is down all the way all way into the tubing and into the compressor If this has happened the fluid sensor has been activated triggered inside the injector to create continuous positive pressure with protectant fluid used to protect the compressor from water damage XENOWORKS DIGITAL MICROINJECTOR OPERATION MANUAL REV 4 13 20100914 21 7 MAINTENANCE 7 1 Regeneration of the Drierite Desiccant The XenoWorks Digital Microinjector needs no routine maintenance except for periodic replacement or regeneration of the Drierite desiccant The indicating Drierite found in the canister on the rear of the microinjector is a desiccant made of calcium sulfate 97 and cobalt chloride 396 Drierite is non toxic and can be handled with few precautions This material is used to remove water vapor from the air intake of the system As it absorbs more moisture it becomes pink in color and must eventually be regenerated dried or replaced with new indicating desiccant canister New canisters are available from Sutter order part number X870700 or the existing granules can be regenerated by following these instructions 1 Turn offthe device and unplug the
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