Freeze-fracture cytochemistry: a new fracture-labeling method for topological analysis of biomembrane molecules.
Freeze-fracture cytochemistry allows visualization of cellular and molecular characteristics of biomembranes in situ. In this review, we discuss freeze-fracture cytochemistry with special reference to a new cytochemical labeling of replicas, the detergent-digestion fracture-labeling technique. In this procedure, unfixed cells are rapidly-frozen, freeze-fractured, and physically stabilized by evaporated platinum/carbon. The frozen cells are then removed from the freeze-fracture apparatus to thaw and are subsequently treated with detergents. After detergent-digestion, replicas are labeled with cytochemical markers. We demonstrate that the technique is a versatile tool for direct analysis of the macromolecular architecture of biomembranes and allows identification of particular intracellular membrane organelles. In addition, we demonstrate the application of ultrasmall gold to freeze-fracture immunocytochemistry. Freeze-fracture cytochemistry is a valuable technique for investigating topology and dynamics of membrane molecules.
Takizawa
T
T
Department of Anatomy, Jichi Medical School, Tochigi, Japan. ttakizawa@jichi.ac.jp
Robinson
J M
JM
D002462:Cell Membrane / Q000378:metabolism; D003902:Detergents; D005614:Freeze Fracturing / Q000379:methods*; D005960:Glucosides; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D008854:Microscopy, Electron / Q000379:methods
D003902:Detergents; D005960:Glucosides; C018619:octyl-beta-D-glucoside