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Ice crystal damage in frozen thin sections: freezing effects and their restoration
Author(s) -
Frederik P. M.,
Busing W. M.
Publication year - 1981
Publication title -
journal of microscopy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.569
H-Index - 111
eISSN - 1365-2818
pISSN - 0022-2720
DOI - 10.1111/j.1365-2818.1981.tb01212.x
Subject(s) - ice crystals , osmium tetroxide , cryoprotectant , microanalysis , osmium , freeze drying , materials science , optical microscope , crystal (programming language) , dry ice , electron microscope , chemistry , mineralogy , scanning electron microscope , cryopreservation , composite material , chromatography , optics , embryo , biochemistry , physics , organic chemistry , ruthenium , computer science , biology , programming language , microbiology and biotechnology , catalysis
SUMMARY Thin sections of unfixed kidney, fast frozen without cryoprotectants, were fixed in osmium tetroxide vapour directly after freeze drying or after 30 min in a moist atmosphere. Dry sections fixed in vapour showed ice crystal damage characteristic for the freezing procedure. This was demonstrated with freeze fracture replicas from the same preparation. Ice crystal holes were obscured in serial sections which were freeze dried and allowed to rehydrate in a moist atmosphere. The same ultrastructural appearance was observed in frozen sections brought to room temperature immediately after cutting. Frozen thin sections from unfixed tissue, if freeze dried, are very sensitive to atmospheric conditions and need some form of stabilization (e.g. osmium vapour fixation, sealing with an evaporated carbon film) before electron microscope images can be interpreted as representative for the frozen state. Restoration of ice crystal damage can occur by melting frozen sections or by rehydration of freeze dried frozen sections. Restoration phenomena will impair studies aimed at the localization of diffusible substances by autoradiography or X‐ray microanalysis.