Correlative super-resolution fluorescence and metal-replica transmission electron microscopy

Nature Methods (Impact Factor: 32.07). 01/2014; 11(3). DOI: 10.1038/nmeth.2816
Source: PubMed


We combine super-resolution localization fluorescence microscopy with transmission electron microscopy of metal replicas to locate proteins on the landscape of the cellular plasma membrane at the nanoscale. We validate robust correlation on the scale of 20 nm by imaging endogenous clathrin (in two and three dimensions) and apply the method to find the previously unknown three-dimensional position of the endocytic protein epsin on clathrin-coated structures at the plasma membrane.

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    • "Thus, the combination of light and EM methods is particularly exciting (de Boer et al., 2015). Several correlative light and EM methods are now bridging the gap and providing the best information from both methods (Kukulski et al., 2012; Sochacki et al., 2014). With these advances it is possible to imagine determining the position of hundreds/thousands of protein types in a cell. "
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