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Freeze‐fracture for scanning electron microscopy
Author(s) -
Haggis G. H.,
PhippsTodd Beverly
Publication year - 1977
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.1365-2818.1977.tb00059.x
Subject(s) - cryoprotectant , scanning electron microscope , electron microscope , fracture (geology) , microscopy , chemistry , biophysics , biology , materials science , pathology , cryopreservation , microbiology and biotechnology , composite material , medicine , optics , embryo , physics
SUMMARY Two different freeze‐fracture methods are explored for preparation of biological material for scanning electron microscopy. In the simpler method the tissues are first fixed and dehydrated. They are then frozen and fractured, and after thawing, critical‐point dried. This method has already been used in a number of studies of animal tissues (heart, liver, kidney). It is applied here to the examination of plant material (leaf mesophyll cells). In the second method tissues, or cells, are first infiltrated with cryoprotectant (dimethylsulphoxide) then frozen and fractured, and not fixed until after thawing. The fixed tissues are finally dehydrated and critical‐point dried. This method also has previously been used in the study of animal tissues, and is applied here to carrot protoplasts, chicken erythrocytes, and leaf mesophyll cells.