A surface replica method: a useful tool for studies of the cytoskeletal network in red cell membranes of normal subjects and patients with a β-spectrin mutant (spectrin Le Puy: β220/214)

Visualization of the components of the red cell membranes, and especially the structure of cytoskeletal proteins in situ, has become a requisite in studies of red cell membrane disorders. There has been a search for a consistent and dependable method for detecting these structures. In the present study, the surface replica method was used with transmission electron microscopy to examine the cytoskeletal network of the red cell ghosts of a normal control and patients with a β-spectrin mutant (β-spectrin Le Puy). The surface replica method is well-suited to observation of the cytoskeletal network of the membranes in a nearly native in situ condition. Immunogold labelling with anti-membrane protein antibodies is easily applicable to the identification of each component of the cytoskeletal proteins. The findings obtained under normal and pathological conditions using the surface replica method corresponded with those made by the quick-freeze, deep-etching method.