作者
Benjamin G Kopek, Maria G Paez-Segala, Gleb Shtengel, Kem A Sochacki, Mei G Sun, Yalin Wang, C Shan Xu, Schuyler B van Engelenburg, Justin W Taraska, Loren L Looger, Harald F Hess
发表日期
2017/5
期刊
Nature protocols
卷号
12
期号
5
页码范围
916-946
出版商
Nature Publishing Group UK
简介
Our groups have recently developed related approaches for sample preparation for super-resolution imaging within endogenous cellular environments using correlative light and electron microscopy (CLEM). Four distinct techniques for preparing and acquiring super-resolution CLEM data sets for aldehyde-fixed specimens are provided, including Tokuyasu cryosectioning, whole-cell mount, cell unroofing and platinum replication, and resin embedding and sectioning. The choice of the best protocol for a given application depends on a number of criteria that are discussed in detail. Tokuyasu cryosectioning is relatively rapid but is limited to small, delicate specimens. Whole-cell mount has the simplest sample preparation but is restricted to surface structures. Cell unroofing and platinum replication creates high-contrast, 3D images of the cytoplasmic surface of the plasma membrane but is more challenging than whole …
引用总数
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