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A copper block method for freezing non‐cryoprotected tissue to produce ice‐crystal‐free regions for electron microscopy
Author(s) -
Dempsey G. P.,
Bullivant S.
Publication year - 1976
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.1365-2818.1976.tb02405.x
Subject(s) - ice crystals , electron microscope , liquid nitrogen , copper , cryofixation , crystal (programming language) , materials science , cryoprotectant , microscopy , chemistry , crystallography , optics , biology , cryopreservation , physics , metallurgy , embryo , organic chemistry , computer science , programming language , microbiology and biotechnology
SUMMARY A wide variety of plant and animal tissues were prepared for electron microscopy by freeze‐substitution, after rapid freezing on a liquid nitrogen cooled copper block by the van Harreveld method. Measurements were made of ice crystal size versus depth in tissues that had not been treated with any cryoprotectants. Ice crystal size increased exponentially with depth. It was confirmed that a narrow surface band (approximately 12 μm) was frozen sufficiently rapidly to prevent the formation of electron microscopically‐visible ice crystals.