Freeze-substitution: Origins and applications

Freeze-substitution is a physicochemical process in which biological specimens are immobilized and stabilized for microscopy. Water frozen within cells is replaced by organic solvents at subzero temperatures. Freeze-substitution is widely used for ultrastructural and immunocytochemical analyses of c...

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Bibliographic Details
Published inInternational Review of Cytology Vol. 206; pp. 45 - 96
Main Author Shiurba, Robert
Format Book Chapter Journal Article
LanguageEnglish
Published United States Elsevier Science & Technology 2001
Subjects
Animals
Biochemistry
Biophysics
Cellular biology
Coloring Agents - metabolism
Cryopreservation
Fixatives - chemistry
Freeze Drying - history
Freeze Substitution - history
Freeze Substitution - instrumentation
Freeze Substitution - methods
Freeze-substitution
History, 19th Century
History, 20th Century
Humans
Immunocytochemistry
Immunohistochemistry - instrumentation
Immunohistochemistry - methods
Microscopy - instrumentation
Microscopy - methods
Phenotype
Solvents - chemistry
Water - chemistry
Cryopreservation
Immunocytochemistry
Freeze-substitution
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Summary:Freeze-substitution is a physicochemical process in which biological specimens are immobilized and stabilized for microscopy. Water frozen within cells is replaced by organic solvents at subzero temperatures. Freeze-substitution is widely used for ultrastructural and immunocytochemical analyses of cells by transmission and scanning electron microscopy. Less well recognized is its superiority over conventional chemical fixation in preserving labile and rare tissue antigens for immunocytochemistry by light microscopy. In the postgenome era, the focus of molecular genetics will shift from analyzing DNA sequence structure to elucidating the function of gene networks, the intercellular effects of polygenetic diseases, and the conformational rearrangements of proteins in situ. Novel strategies will be needed to integrate knowledge of chemical structures of normal and abnormal macromolecules with the physiology and developmental biology of cells and tissues from whole organisms. This review summarizes the progress and future prospects of freeze-substitution for such explorations.
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ISBN:0123646103
9780123646101
ISSN:0074-7696
2163-5854
DOI:10.1016/S0074-7696(01)06019-3