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Method to extract minimally damaged collagen fibrils from tendon
Author(s) -
Yehe Liu,
Nelly AndarawisPuri,
Steven J. Eppell
Publication year - 2016
Publication title -
journal of biological methods
Language(s) - English
Resource type - Journals
ISSN - 2326-9901
DOI - 10.14440/jbm.2016.121
Subject(s) - fibril , collagen fibril , tendon , electron microscope , dermis , extraction (chemistry) , scanning electron microscope , chemistry , materials science , microscopy , biophysics , anatomy , chromatography , optics , biology , composite material , biochemistry , physics
A new method is presented to extract collagen fibrils from mammalian tendon tissue. Mammalian tendons are treated with a trypsin-based extraction medium and gently separated with tweezers in an aqueous solution. Collagen fibrils released in the solution are imaged using both dark-field light microscopy and scanning electron microscopy. The method successfully extracts isolated fibrils from rat tail and patellar tendons. To examine whether the method is likely to damage fibrils during extraction, sea cucumber dermis fibril lengths are compared against those obtained using only distilled water. The two methods produce fibrils of similar lengths. This is contrasted with fibrils being shortened when extracted using a tissue homogenizer. Scanning electron microscopy shows the new method preserves D-banding features on fibril surfaces and that fibril diameter does not vary substantially compared with water extracted fibrils.

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