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从肌腱中提取最小损伤胶原纤维的方法。

Method to extract minimally damaged collagen fibrils from tendon.

作者信息

Liu Yehe, Andarawis-Puri Nelly, Eppell Steven J

机构信息

Department of Biomedical Engineering, Case Western Reserve University, Cleveland, OH 44106, USA.

Leni and Peter W. May Department of Orthopaedics, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.

出版信息

J Biol Methods. 2016 Sep 16;3(4):e54. doi: 10.14440/jbm.2016.121. eCollection 2016.

Abstract

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.

摘要

提出了一种从哺乳动物肌腱组织中提取胶原纤维的新方法。用基于胰蛋白酶的提取介质处理哺乳动物肌腱,并在水溶液中用镊子轻轻分离。使用暗场光学显微镜和扫描电子显微镜对溶液中释放的胶原纤维进行成像。该方法成功地从大鼠尾巴和髌腱中提取出分离的纤维。为了检验该方法在提取过程中是否可能损坏纤维,将海参真皮纤维的长度与仅使用蒸馏水获得的纤维长度进行比较。两种方法产生的纤维长度相似。这与使用组织匀浆器提取时纤维缩短形成对比。扫描电子显微镜显示,新方法保留了纤维表面的D带特征,并且与水提取的纤维相比,纤维直径没有显著变化。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fcf8/6706113/24997d34dbf6/jbm-3-4-e54-g001.jpg

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