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Localization of fluorescently labeled structures in frozen-hydrated samples using integrated light electron microscopy

Item Type:Article
Title:Localization of fluorescently labeled structures in frozen-hydrated samples using integrated light electron microscopy
Creators Name:Faas, F.G.A. and Bárcena, M. and Agronskaia, A.V. and Gerritsen, H.C. and Moscicka, K.B. and Diebolder, C.A. and van Driel, L.F. and Limpens, R.W.A.L. and Bos, E. and Ravelli, R.B.G. and Koning, R.I. and Koster, A.J.
Abstract:Correlative light and electron microscopy is an increasingly popular technique to study complex biological systems at various levels of resolution. Fluorescence microscopy can be employed to scan large areas to localize regions of interest which are then analyzed by electron microscopy to obtain morphological and structural information from a selected field of view at nm-scale resolution. Previously, an integrated approach to room temperature correlative microscopy was described. Combined use of light and electron microscopy within one instrument greatly simplifies sample handling, avoids cumbersome experimental overheads, simplifies navigation between the two modalities, and improves the success rate of image correlation. Here, an integrated approach for correlative microscopy under cryogenic conditions is presented. Its advantages over the room temperature approach include safeguarding the native hydrated state of the biological specimen, preservation of the fluorescence signal without risk of quenching due to heavy atom stains, and reduced photo bleaching. The potential of cryo integrated light and electron microscopy is demonstrated for the detection of viable bacteria, the study of in vitro polymerized microtubules, the localization of mitochondria in mouse embryonic fibroblasts, and for a search into virus-induced intracellular membrane modifications within mammalian cells.
Keywords:Correlative Microscopy, Correlative Light Electron Microscopy, Integrated Light Electron Microscopy, Cryo Correlative Microscopy, Animals, Chlorocebus aethiops
Source:Journal of Structural Biology
Page Range:283-290
Date:March 2013
Official Publication:https://doi.org/10.1016/j.jsb.2012.12.004
PubMed:View item in PubMed

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