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A Single Method for Cryofixation and Correlative Light, Electron Microscopy and Tomography of Zebrafish Embryos
Author(s) -
Nixon Susan J.,
Webb Richard I.,
Floetenmeyer Matthias,
Schieber Nicole,
Lo Harriet P.,
Parton Robert G.
Publication year - 2009
Publication title -
traffic
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.677
H-Index - 130
eISSN - 1600-0854
pISSN - 1398-9219
DOI - 10.1111/j.1600-0854.2008.00859.x
Subject(s) - immunogold labelling , cryofixation , zebrafish , biology , fluorescence , green fluorescent protein , electron microscope , biophysics , live cell imaging , microbiology and biotechnology , correlative , fluorescence microscope , ultrastructure , microscopy , in situ , vitrification , anatomy , cell , biochemistry , chemistry , pathology , optics , physics , medicine , linguistics , philosophy , andrology , organic chemistry , gene
The zebrafish is a powerful vertebrate system for cell and developmental studies. In this study, we have optimized methods for fast freezing and processing of zebrafish embryos for electron microscopy (EM). We show that in the absence of primary chemical fixation, excellent ultrastructure, preservation of green fluorescent protein (GFP) fluorescence, immunogold labelling and electron tomography can be obtained using a single technique involving high‐pressure freezing and embedding in Lowicryl resins at low temperature. As well as being an important new tool for zebrafish research, the maintenance of GFP fluorescence after fast freezing, freeze substitution and resin embedding will be of general use for correlative light and EM of biological samples.