An Imaging Protocol to Quantitate Microvilli Ultrastructure Induction by Epithelia.

Le résumé fourni par la source

Current methodology to quantitatively study microvilli induction by epithelia involves analysis by transmission electron microscopy \(TEM) and measurement of microvilli length perpendicular to the membrane.Since this technique involves cutting ultrathin sections of the cell or tissue, the entire surface area of the membrane is not covered and would require a large number of ultrathin sections to reconstruct the surface in order to accurately determine microvilli status.Here we describe a protocol that takes advantage of scanning electron microscopy \(SEM) to cover the entire epithelial cell surface of groups of cells and reveals detailed ultrastructure of microvilli.These images can be quantitated using Nikon Imaging Software \(NIS) that distinguishes microvilli structures, and even substructures, from at apical membrane surfaces.The protocol described constitutes a robust assay to quantitatively analyse microvilli generated by cultured epithelia but can potentially also be applied to epithelial tissues and potentially other tissues and cell types that generate microvilli-like structures.

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