title: Freeze-substitution: origins and applications.
abstract: 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.
authors.affiliation: Misato Inc., Satte-shi, Saitama, Japan.
authors.forename: R
authors.initials: R
authors.lastname: Shiurba
mesh_terms: D000818:Animals; D004396:Coloring Agents / Q000378:metabolism; D005404:Fixatives / Q000737:chemistry; D005612:Freeze Drying / Q000266:history; D017110:Freeze Substitution / Q000266:history / Q000295:instrumentation / Q000379:methods*; D049672:History, 19th Century; D049673:History, 20th Century; D006801:Humans; D007150:Immunohistochemistry / Q000295:instrumentation / Q000379:methods*; D008853:Microscopy / Q000295:instrumentation / Q000379:methods; D010641:Phenotype; D012997:Solvents / Q000737:chemistry; D014867:Water / Q000737:chemistry
chemical_list: D004396:Coloring Agents; D005404:Fixatives; D012997:Solvents; D014867:Water