Preparation of basal cell membranes for scanning probe microscopy

• Published in: FEBS letters Vol. 436; no. 2; pp. 179 - 184
• Main Authors: Ziegler, U, Vinckier, A, Kernen, P, Zeisel, D, Biber, J, Semenza, G, Murer, H, Groscurth, P
• Format: Journal Article
• Language: English
• Published: England Elsevier B.V 02.10.1998
• Subjects: AFM, atomic force microscopy; Animals; Atomic force microscopy; Basal cell membrane; Cell Line; Cell Membrane - ultrastructure; Confocal laser scanning microscopy; Cytoskeleton; Dogs; FITC, fluorescein-5-isothiocyanate; Kidney; MDCK, Madine-Darby canine kidney; Microscopy, Atomic Force - methods; Microscopy, Electron; Microscopy, Fluorescence; PBS, phosphate buffered saline; Scanning probe microscopy; Sensitivity and Specificity; SNOM, scanning near field optical microscopy; SPM, scanning probe microscopy; Atomic force microscopy; SNOM, scanning near field optical microscopy; SPM, scanning probe microscopy; Scanning probe microscopy; Basal cell membrane; MDCK, Madine-Darby canine kidney; PBS, phosphate buffered saline; Cytoskeleton; Confocal laser scanning microscopy; AFM, atomic force microscopy; FITC, fluorescein-5-isothiocyanate
• ISSN: 0014-5793; 1873-3468
• DOI: 10.1016/S0014-5793(98)01118-1

Scanning probe microscopy has the potential for investigating membranes in a physiological environment. We prepared with a lysis-squirting protocol basal cell membranes, that are suitable for scanning probe microscopy. Investigations using atomic force microscopy under liquid revealed cellular filaments which correlated perfectly with fluorescently stained actin filaments. Globular structures with a diameter as little as 10 nm could be resolved by stripping cytoplasmic components from the membranes. Therefore, cytoplasmic sides of supported basal cell membranes prove useful to gain high resolution with scanning probe microscopy in studies of plasma membrane associated structures and processes under buffer solution.