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High-pressure Freezer &
Automated Freeze-substitution System

Standard vs HPF, yeastHPM100 instrumentAutomated Freeze Substitution
click image for larger version

Model Information

  • High-pressure Freezer: Leica HPM-100
  • Automated Freeze-substitution System: Leica AFS-2
  • Cite in your manuscripts: NIH Grant S10 RR026445 (Institute:Research Resources [RR])
    Installed: Mar 2011

Specifications

  • Freeze specimen under 20atm within milliseconds! Prevent formation of ice-crystals that damage cell membranes.
     Freezing Curve
  • When frozen, crosslinking agents, particularly membrane-stabilizing reagents (e.g. osmium tetroxide), still work! Preservation is excellent!
  • Correlative light/fluorescence microscopy:
    • Grow cells on special sapphire coverslips.
    • Observe cells on light/fluorescence/confocal microscope (your choice!)
    • After walking down the hall, freeze within 10s!
    • After processing, compare light/fluorescence images with EM.

Applications

  • Preserving delicate membrane morphology:
    • nuclear membrane, nuclear pores
    • endoplasmic reticulum, both tubule, cisternae and plasma-membrane-associated ER (tubER, cecER, PM-ER)
  • Preserving large structures:
    • cell clusters (bacteria/biofilms)
    • whole Drosophila embryos

Examples

HPF yeast
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Standard vs HPF, Drosophila
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