Journal
TRAFFIC
Volume 10, Issue 2, Pages 131-136Publisher
WILEY
DOI: 10.1111/j.1600-0854.2008.00859.x
Keywords
CLEM; cryofixation; GFP; tomography; zebrafish
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Funding
- Australian Microscopy and Microanalysis Research Facility (AMMRF)
- NCRIS Australian Research Council initiative
- Australian Cancer Research Foundation
- Australian Phenomics Facility
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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.
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