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High-pressure Freezer &
Automated Freeze-substitution System
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- High-pressure Freezer: Leica HPM-100
- Automated Freeze-substitution System: Leica AFS-2
- Freeze specimen under 20atm within milliseconds! Prevent formation of ice-crystals that damage cell membranes.
- 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.
- 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