Difference: InsertCryoSample (2 vs. 3)

Revision 305 Oct 2006 - Main.KdDerr

 
META TOPICPARENT name="TecnaiInstructions"
Changed:
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<		Return to Cryo EM Website at http://www.nysbc.org/facilities/CEM

Cryo sample loading

Contents

Apertures

  • Condenser aperture #2
  • Objective aperture #4

Check for beam before loading sample

  • Insert specimen holder to first stop
  • In setup menu "open column valves"
  • If no beam lower magnification
  • In low dose menu Lexposure mode, spot size should read 2 if not reload good configuration (i.e. JB1) save as your own
  • Check alignment of scope before inserting sample

Loading cryo sample into holder

  • Reset holder
  • Set holder value ? to -79
  • Top off LN2 in cold trap
  • Retrieve sample
  • Practice removing clip w/o LN2
  • Cool down sample dewer and sample loading end (fill about ½ with LN2)
  • After boiling subsides top off
  • Remove grid
  • Place in holder
  • Replace clip ring
  • Press firm with sample tap
    • Keep 2mm of LN2 covering sample at all times

Loading cryo holder into scope

  • Insert sample to first stop (do not let go until vacuum kicks in)
  • Identify holder at the bottom of the screen
  • When light goes out, push Reset holder
  • Hold sample turn upright and insert slowly
  • Top off dewar
  • Clear with blow off pipe to calm LN2
  • Cover Dewar
  • Wait one hour before imaging to achieve equilibrium temperature with no gradients

Finally

  • Open shutter and move sample to make sure vacuum does not fluctuate before opening column valves

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

-- DavidStokes - 18 Mar 2005

>
>		Return to Cryo EM Website at http://www.nysbc.org/facilities/CEM

Cryo sample loading

Contents

Apertures

  • Condenser aperture #2
  • Objective aperture #4

Check for beam before loading sample

  • Insert specimen holder to first stop
  • In setup menu "open column valves"
  • If no beam lower magnification
  • In low dose menu Lexposure mode, spot size should read 2 if not reload good configuration (i.e. JB1) save as your own
  • Check alignment of scope before inserting sample

Loading cryo sample into holder

  • Reset holder
  • Set holder alpha value to -79
  • Top off LN2 in cold trap
  • Retrieve sample
  • Practice removing clip w/o LN2
  • Cool down sample dewer and sample loading end (fill about ½ with LN2)
  • After boiling subsides top off
  • Remove grid
  • Place in holder
  • Replace clip ring
  • Press firm with sample tap
    • Keep 2mm of LN2 covering sample at all times

Loading cryo holder into scope

  • Insert sample to first stop (do not let go until vacuum kicks in)
  • Identify holder at the bottom of the screen
  • When light goes out, push Reset holder
  • Hold sample turn upright and insert slowly
  • Top off dewar
  • Clear with blow off pipe to calm LN2
  • Cover Dewar
  • Wait one hour before imaging to achieve equilibrium temperature with no gradients

Finally

  • Open shutter and move sample to make sure vacuum does not fluctuate before opening column valves

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

-- DavidStokes - 18 Mar 2005

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