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Protocols
cryosectioning,
avidin-biotin, sample fixation |
Buffers
tissue fixation, permeabilization,
PBS ... |
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Links
to
microscopy and histochemistry resources
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| cryosectioning |
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1.
Cool the cryostat to approximately -20 C (if it
is too cold, the tissue will be difficult to cut)
2. create a small bowl shaped aluminum foil basket
to hold the tissue sample (see images)
3. pour some cryomatrix embedding agent into the
foil and place the tissue sample into the liquid
matrix
4. freeze the sample by dipping the foil into liquid
nitrogen (use forceps to hold the foil)
5. place the frozen tissue/ cryomatrix block on
the 'chuck' that guides the sample against the cryostat
blade (adhere with a dab of cryomatrix solution
6. once the tissue block has frozen into place,
move the block into place and cut the sections to
the desired thickness (7 - 20 microns)
7. pick each slice up by pressing a glass microscope
slide on the tissue so that it melts/adheres
8. leave each microscope slide containing tissue
section in the cryostat chamber in a microscope
rack to keep them frozen while you cut the rest
of the block
9. wrap the rack in aluminium foil and store at
-80 C
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