![]() We describe how to use 3View for studying collagen fibril organization in vivo and show how to find and track individual fibrils. The protocol described below is suitable for preparing tissues for TEM and SBF-SEM (by 3View). However, serial block face-scanning electron microscopy (SBF-SEM) has emerged as a time-efficient alternative to ssTEM. Serial-section TEM (ssTEM) has been used to visualize fibril 3D organization in vivo. Here we describe a protocol for measuring fibril diameter as well as fibril volume fraction, mean fibril length, fibril cross-sectional shape and fibril 3D organization, all of which are major determinants of tissue function. Transmission electron microscopy (TEM) is used to assess the formation of the fibrils, predominantly by measuring fibril diameter. Abnormal fibril assembly and organization results in scarring, fibrosis, poor wound healing and connective tissue diseases. ![]() Collagen fibrils are the major tensile element in vertebrate tissues, in which they occur as ordered bundles in the extracellular matrix.
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