Difference: HpfUse (7 vs. 8)

Revision 802 Oct 2009 - Main.KdDerr

 
META TOPICPARENT name="HighPressFreezer"
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Return to Cryo EM web page at http://www.nysbc.org/facilities/CEM
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Principles & Protocols



Protocols - Using the Bal-Tec High Pressure Freezer

Jump to Principles

 
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Return to Cryo EM website at http://www.nysbc.org/facilities/CEM
 
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Using the Bal-Tec High Pressure Freezer

 
  • If you are having problems turn the HPF off and restart
  • Download MSWord version of these procedures: HPF-procedures.doc
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START-UP

 
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Contents

STARTUP

 
  • 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
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    • Make sure planchet is loaded in measuring holder
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    • 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

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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