Difference: HpfUse (9 vs. 10)

Revision 1018 Jan 2013 - Main.MarienaSilvestry

 
META TOPICPARENT name="HighPressFreezer"
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Principles & Protocols

Protocols - Using the Bal-Tec High Pressure Freezer

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  • If you are having problems turn the HPF off and restart
  • Download MSWord version of these procedures: HPF-procedures.doc
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Notes from repair done in the Fall of 2012 to the measuring holder:

In late Fall 2012 Bill Graham from Bibst Labs came to service the HPM 010 (Bal-Tac). He serviced the machine, changed the exhaust baffles on the machine (put 2 new in, old ones were cracked) and also serviced the heads on both the measuring and the sample holder. Both holders have new heads and the error message which the machine was previously showing was gone, therefore it is now possible to measure both pressure and temperature maintenance, which we couldn't do before.

Bill also created an operation manual/list for this machine, which is attached below. Mariena made new log sheets to keep track of the shots fired, samples frozen, etc. Shots are measured on the side panel of the machine, and you need a key to open the panel. Both Kent McDonald? and Bill Graham recommend getting the machine serviced every 1000 shots.

A copy of Bill's instructions on how to operate the machine can be found also on the clipboard where the shot log is. The old instructions are also there. A copy of the report is attached (see below) and can also be found on the HPF binder above Minotaur (Nicolas' computer).

* HPM_010_repair_report_10_12_12.pdf: Repair Report Fall 2012

* HPM_010_User_NYSBC.pdf: HPM 010 Manual

 

START-UP

  • Fill water recirculator
  • Turn on 2 power switches 1hr before use (objecthead temperature must read 40 b4 use)
  • Fill with 2-propanol
  • Turn on N2 tank (meter should read between 4-5 bar)
  • Purge system with N2 for 30-40 min (air heater button)
  • Turn on LN2 tank
  • Push START
  • Push NITROGEN
    • If no noise from LN2, tap gently on solenoid valve inside left side
  • Wait until red NITROGEN light goes out
  • Push DRIVE IN
  • Wait until drive in light goes out (pressure should increase to ~200 bar)
  • Push AUTO (pressure should increase to 310 bar and subsequently cycle 295-310)
  • Record freezing parameters with measuring holder before commencing
    • Make sure planchette is loaded in measuring holder
    • Pressure Maintenance should remain >0.5 sec, or grease/change o-ring
    • Use laptop and Matlab to record analogue trace of pressue and temperature
    • Save pdf of trace on laptop

CHANGE LN2 TANK DURING RUN

  • Push STOP
  • Warm up LN2 connection with heat gun
  • Change tank quickly, especially when humid
  • Push START
  • Push NITROGEN
  • Wait until red NITROGEN light goes out
  • Push AUTO (drive in not necessary)

SHUTDOWN

  • Push STOP
  • Turn off LN2 tank
  • Turn off N2 tank
  • Release pressure at 2-propanol inlet
  • Turn off key switch
  • Leave on Mains switch this will continue recirculating the water for 10 hours before shutdown
    • make sure water bath is full of water
  • Remove holder from Objective head and put on black plastic cap

Visit Cryo EM website at http://www.nysbc.org/facilities/CEM

-- DavidStokes - 18 Mar 2005

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META FILEATTACHMENT attr="h" comment="" date="1111119919" name="HPF-procedures.doc" path="HPF-procedures.doc" size="20992" user="DavidStokes" version="1.1"
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META FILEATTACHMENT attr="" autoattached="1" comment="Repair Report Fall 2012" date="1358544381" name="HPM_010_repair_report_10_12_12.pdf" path="HPM_010_repair_report_10_12_12.pdf" size="57743" user="Main.MarienaSilvestry" version="1"
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META FILEATTACHMENT attr="" autoattached="1" comment="HPM 010 Manual" date="1358544403" name="HPM_010_User_NYSBC.pdf" path="HPM_010_User_NYSBC.pdf" size="167554" user="Main.MarienaSilvestry" version="1"
META FILEATTACHMENT attr="h" autoattached="1" comment="" date="1144161563" name="HPF-procedures.doc" path="HPF-procedures.doc" size="20992" user="DavidStokes" version="1.1"
 
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