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EF600 Instruction Manual

Asymptote Ltd.

St John’s Innovation Centre, Cowley Road, Cambridge, CB4 0WS, UK

Tel: +44 1223 421161, Fax: +44 1223 421166, E-mail: [email protected]

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7.

References

Kim SJ, Park JH, Lee JE, Kim JM, Lee, JB, Moon, SY, Roh SI, Kim CG and
Yoon, HS (2004) Effects of type IV collagen and laminin on the
cryopreservation of human embryonic stem cells.

Stem Cells

22

, 950-961.

Lasalle P, Testart J and Renard JP (1985) Human embryo features that
influence the success of cryopreservation with he use of 1, 2 propanediol.

Fertility and Sterility

44

645-651.

Morris GJ, Acton E, and Avery S

(1999)

A novel approach to sperm

cryopreservation.

Human Reproduction

14

1013-1021

.

Appendix 1: Troubleshooting

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Summary of Contents for EF600 Series

Page 1: ...ries Instruction Manual Asymptote nitrogen free controlled rate freezer Asymptote Ltd St John s Innovation Centre Cowley Road Cambridge CB4 0WS Tel 44 1223 421161 Fax 44 1223 421166 e mail info asymptote co uk ...

Page 2: ...MES EF600 01 EF600 10 11 5 OPERATING PROCEDURE FOR THE EF600 FREEZER 13 5 1 PRE OPERATION CHECKS 13 5 2 PREPARING THE NUCLEATING TOOL 13 5 3 PREPARING THE STRAWS 13 5 4 LOADING THE STRAWS 13 5 5 STARTING THE DATA LOGGING EF600 01 EF600 10 14 5 6 STARTING THE FREEZING CYCLE 14 5 7 SAMPLE NUCLEATION 15 5 8 END OF CYCLE 15 5 9 STOPPING THE DATA LOGGING 16 5 10 RESTARTING THE CYCLE 16 5 11 CLEANING TH...

Page 3: ...IX 1 TROUBLESHOOTING 21 A1 1 FREEZING CYCLE WILL NOT START 22 A1 2 NO POWER TO UNIT 22 A1 3 DATA LOGGING DOES NOT FUNCTION 22 A1 4 STRAWS STUCK TO SAMPLE PLATE 22 APPENDIX 2 INSTALLING ASYMPTOTE S DATA LOGGING SOFTWARE 24 A2 1 SYSTEM REQUIREMENTS 24 A2 2 INSTALLING THE PROGRAM 24 APPENDIX 3 UNINTERRUPTIBLE POWER SUPPLY UPS 29 APPENDIX 4 CRYOPEN NUCLEATING TOOL OPERATION 29 ...

Page 4: ...man embryo freezing cycle 17 Figure 6 2 Pre programmed human oocyte freezing cycle 18 Figure 6 3 Pre programmed human blastocyst freezing cycle 18 Figure 6 4 Pre programmed human embryonic stem cell freezing cycle 19 Figure 6 5 Pre programmed human sperm freezing cycle 19 Figure 6 6 Pre programmed human spermatozoa non linear freezing cycle 20 Figure A2 1 Installation box 1 24 Figure A2 2 Installa...

Page 5: ...time of ordering EF600 10 Freezer with data logging to a dedicated PC supplied by Asymptote Ltd The EF600 10 is pre loaded with six standard freezing programs embryo oocyte blastocyst stem cell spermatozoa linear program and spermatozoa non linear program which the user selects in the supplied data logging software Additional programs up to a total of 10 may be specified at the time of ordering Th...

Page 6: ...te during and after the freezing cycle to avoid cold burns Do not move the freezer whilst operating as the baseplate will be hot Operating the cryopen nucleating tool Read the instruction manual and safety instructions supplied with the cryopen nucleating tool Ensure that gloves are worn during use and never point the cryopen towards the face Plate temperature The plate temperature should never ex...

Page 7: ...munications cable e Laptop computer with data logging software for either EF600 01 or EF600 10 installed f CD back up of installed software Optional extras such as a cryopen nucleating tool and UPS will have been supplied with the EF600 freezer if ordered If the unit shows any sign of damage or if any of the items listed above are missing please contact the supplier immediately 2 2 Positioning the...

Page 8: ...ions lead 9 pin D type male to female between com port 1 the serial port on the computer and the back of the freezer Power up the freezer unit using the ON OFF switch on the front panel of the unit figure 2 4 The switch illuminates red when the power is switched on Figure 2 4 Front panel of freezer unit keypad and ON OFF switch 3 Starting Asymptote s Data Logging Software EF600 01 EF600 10 ...

Page 9: ...e shortcut icon on your desktop figure 3 or by selecting Asymptote Data Logger from the program list in the start menu on your computer Figure 3 Desktop shortcut icon The program window will open Initial values for the Set Point Temperature Plate Temperature and Stirling Voltage V will be displayed as 999 00 these will automatically update once communications between the freezer and the computer h...

Page 10: ...temperature of the sample plate The current Stirling Voltage V is displayed in green on the right of the screen This is the output level of the Stirling Engine and is only provided for diagnostic engineering purposes only The graph can show either the last minute of the cycle or the whole run Check last minute or whole run to switch between these features The time elapsed and the total run time of...

Page 11: ... 10 When communications have been established user interface is no longer reading 999 select the logging interval and the desired freezing program from the drop down menus figure 4 1 10 seconds is suggested as a default for the logging interval Figure 4 1 Freezing profile 4 2 Log file names EF600 01 EF600 10 Choose profile EF600 10 Choose logging interval EF600 01 EF600 10 ...

Page 12: ...m is not logging by clicking on the grey SPECIFY FILE PATH button Choose a clear and distinguishable file name for the logged data Do not include or any other punctuation marks in the file name Do not include a file name extension The default extension is csv Files are also automatically logged as encrypted files with the extension aef If a file already exists with the file name chosen any logged ...

Page 13: ...ive tools for nucleating e g forceps may be used if sterility of the operating environment is not essential These should be placed in liquid nitrogen before the freezing cycle is started 5 3 Preparing the straws Prepare up to 12 0 25ml standard IMV straws as normal and seal with a straw plug crytaseal PVA powder or other approved sealant Thoroughly dry the outside of the straws after preparation t...

Page 14: ... has been given To go back and check press Cancel to start logging click on OK 5 6 Starting the freezing cycle When the straws have been positioned correctly wait for at least one minute before starting the freezing cycle to allow the straws to reach thermal equilibrium with the plate Note that the freezer must also have been switched on for at least two minutes in order for the controller to init...

Page 15: ... nucleated unless otherwise specified Remove the lid of the freezer completely Touch the nucleating tool if using the cryopen please refer to Appendix 4 for operating instructions to the first straw for approximately 1 second figure 5 7 Repeat with the remaining straws and close the lid of the freezer Turn off the buzzer by pressing the Buzzer Off button Inspect the straws approximately one minute...

Page 16: ...e available options Alternatively the file may be opened directly from Excel The encrypted file aef extension should be saved separately in an audit folder 5 10 Restarting the cycle The freezer must be switched off and switched back on again before another freezing cycle can run Switch the freezer off using the ON OFF switch at the front of the freezer leave the lid on and wait for the plate to wa...

Page 17: ...00 and EF600 01 will run the standard profile for human embryo cryopreservation section 6 1 The EF600 10 freezer is loaded with all six profiles below sections 6 1 to 6 6 6 1 Human embryo cryopreservation Start temperature 20 C Cool from 20 C to 7 C at 2 C min 1 Hold for nucleation at 7 C for 10 minutes Cool from 7 C to 30 C at 0 3 C min 1 Cool from 30 C to 100 C at 10 C min 1 Hold at 100 C 100 80...

Page 18: ... min 1 Hold at 100 C 100 80 60 40 20 0 20 00 00 00 20 00 40 01 00 01 20 01 40 02 00 Time hh mm Temperature C Figure 6 2 Pre programmed human oocyte freezing cycle 6 3 Human blastocyst cryopreservation Start temperature 20 C Cool from 20 C to 7 C at 1 C min 1 Hold for nucleation at 7 C for 10 minutes Cool from 7 C to 30 C at 0 3 C min 1 Hold at 30 C 30 20 10 0 10 20 00 00 00 20 00 40 01 00 01 20 01...

Page 19: ... Hold at 80 C 80 60 40 20 0 20 00 00 00 20 00 40 01 00 01 20 01 40 02 00 Time hh mm Temperature C Figure 6 4 Pre programmed human embryonic stem cell freezing cycle 6 5 Human spermatozoa cryopreservation linear cooling Start temperature 20 C Cool from 20 C to 7 C at 2 C min 1 Hold for nucleation at 7 C for 10 minutes Cool from 7 C to 100 C at 10 C min 1 Hold at 100 C 100 80 60 40 20 0 20 00 00 00 ...

Page 20: ...spermatozoa cryopreservation non linear cooling Start temperature 20 C Cool from 20 C to 7 C at 2 C min 1 Hold for nucleation at 7 C for 10 minutes Cool non linear from 7 C to 100 C at an approximate rate of 10 C min 1 Hold at 100 C 100 80 60 40 20 0 20 00 00 00 10 00 20 00 30 00 40 Time hh mm ss Temperature C Figure 6 6 Pre programmed human spermatozoa non linear freezing cycle ...

Page 21: ...nd Yoon HS 2004 Effects of type IV collagen and laminin on the cryopreservation of human embryonic stem cells Stem Cells 22 950 961 Lasalle P Testart J and Renard JP 1985 Human embryo features that influence the success of cryopreservation with he use of 1 2 propanediol Fertility and Sterility 44 645 651 Morris GJ Acton E and Avery S 1999 A novel approach to sperm cryopreservation Human Reproducti...

Page 22: ...the power lead is plugged into a socket and that the socket is switched on The protective fuse in the EF600 unit has blown Get a qualified person to check and replace the fuse in the power plug at the rear of the EF600 A1 3 Data logging does not function Communications lead between the EF600 unit and the computer is not properly connected Check the connections at both ends The communications lead ...

Page 23: ...eezing section 6 4 Straws not dried before placing in grooves Dry straws before freezing section 6 3 Note that the grooves fit tightly around the straws such but it should always be possible to remove the straws using tweezers For help with any other problems please contact Asymptote Ltd Asymptote Ltd St John s Innovation Centre Cowley Road Cambridge CB4 0WS UK Tel 44 0 1223 421161 Fax 44 0 1223 4...

Page 24: ... 1 1 To see if your computer requires this non critical update please visit Windows update at www windowsupdate com and scan for ALL available updates d A serial port connection lead 9 pin D type male to female e The software installation CD A2 2 Installing the program Insert the CD and wait for the prompts If the program does not open automatically open the CD using Windows Explorer and double cl...

Page 25: ...up Wizard will open to guide you through the steps required to install the software Select Next to move on from the second installation box Figure A2 2 Installation box 2 The third installation box figure 3 2 3 is a reminder about NET Framework 1 1 If this update has already been installed on your computer select Next to move to the next window Figure A2 3 Installation box 3 ...

Page 26: ...ll logged files and user profiles will be stored in this installation folder We recommend that you install the program in the default installation folder C Program Files Asymptote Asymptote Data Logger Select Browse to choose a different location or select Next to continue Figure A2 4 Installation box 4 Select Next to confirm the installation or Cancel to exit This is the final step before the pro...

Page 27: ...6 E mail info asymptote co uk 27 The Setup Wizard will then install the software The progress bar across the centre of Installation box 6 indicates how long the installation will take Figure A2 6 Installation box 6 When the installation has finished Installation box 7 will be displayed Select Close to exit Figure A2 7 Installation box 7 ...

Page 28: ...ation Centre Cowley Road Cambridge CB4 0WS UK Tel 44 1223 421161 Fax 44 1223 421166 E mail info asymptote co uk 28 The final indication that installation has been successful will be a small message box saying Setup succeeded Figure A2 8 Installation box 8 ...

Page 29: ...ing cycle will still run Connect the power lead from the rear of the freezer unit into the UPS and connect the UPS into the mains supply Appendix 4 Cryopen Nucleating Tool Operation Read the instruction manual supplied with the cryopen before use The cryopen is a medical device that can be used to nucleate the sample straws Connect the cartridge to the cryopen according to the instruction manual T...

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