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Sample preparation and imaging of erythrocyte cytoskeleton with the atomic force microscopy.

Authors
  • Liu, Fei
  • Burgess, Joel
  • Mizukami, Hiroshi
  • Ostafin, Agnes
Type
Published Article
Journal
Cell biochemistry and biophysics
Publication Date
Jan 01, 2003
Volume
38
Issue
3
Pages
251–270
Identifiers
PMID: 12794267
Source
Medline
License
Unknown

Abstract

A novel method for the covalent attachment of erythrocytes to glass microscope coverslips that can be used to image intact cells and the cytoplasmic side of the cell membrane with either solid or liquid mode atomic force microscopy (AFM) is described. The strong binding of cells to the glass surface is achieved by the interaction of cell membrane carbohydrates to lectin, which is bound to N-5-azido-2-nitrobenzoyloxysuccinimide (ANBNOS)-coated coverslips (1). The effectiveness of this method is compared with the other commonly used methods of immobilizing intact erythrocytes on glass coverslips for AFM observations. Experimental conditions of AFM imaging of biologic tissue are discussed, and typical topographies of the extracellular and the cytoplasmic surfaces of the plasma membrane in the dry state and in the liquid state are presented. Comparison of the spectrin network of cell age-separated erythrocytes has demonstrated significant loss in the network order in older erythrocytes. The changes are quantitatively described using the pixel height histogram and window size grain analysis.

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