High-quality ultrastructural preservation using cryofixation for 3D electron microscopy of genetically labeled tissues.

ABSTRACT: Electron microscopy (EM) offers unparalleled power to study cell substructures at the nanoscale. Cryofixation by high-pressure freezing offers optimal morphological preservation, as it captures cellular structures instantaneously in their near-native state. However, the applicability of cryofixation is limited by its incompatibility with diaminobenzidine labeling using genetic EM tags and the high-contrast en bloc staining required for serial block-face scanning electron microscopy (SBEM). In addition, it is challenging to perform correlated light and electron microscopy (CLEM) with cryofixed samples. Consequently, these powerful methods cannot be applied to address questions requiring optimal morphological preservation. Here, we developed an approach that overcomes these limitation

SUBMITTER: Tsang TK 

PROVIDER: S-EPMC5988420 | biostudies-literature | 2018 May

REPOSITORIES: biostudies-literature