Correlative light and electron microscopy (CLEM) as a tool to visualize microinjected molecules and their eukaryotic sub-cellular targets.

Journal of Visualized Experiments : JoVE
L Evan Reddick, Neal M Alto

Abstract

The eukaryotic cell relies on complex, highly regulated, and functionally distinct membrane bound compartments that preserve a biochemical polarity necessary for proper cellular function. Understanding how the enzymes, proteins, and cytoskeletal components govern and maintain this biochemical segregation is therefore of paramount importance. The use of fluorescently tagged molecules to localize to and/or perturb subcellular compartments has yielded a wealth of knowledge and advanced our understanding of cellular regulation. Imaging techniques such as fluorescent and confocal microscopy make ascertaining the position of a fluorescently tagged small molecule relatively straightforward, however the resolution of very small structures is limited. On the other hand, electron microscopy has revealed details of subcellular morphology at very high resolution, but its static nature makes it difficult to measure highly dynamic processes with precision. Thus, the combination of light microscopy with electron microscopy of the same sample, termed Correlative Light and Electron Microscopy (CLEM), affords the dual advantages of ultrafast fluorescent imaging with the high-resolution of electron microscopy. This powerful technique has been imp...Continue Reading

References

Jun 26, 2008·Histochemistry and Cell Biology·Ben N G Giepmans
Jan 13, 2009·Annals of Anatomy = Anatomischer Anzeiger : Official Organ of the Anatomische Gesellschaft·Terry M MayhewMatthias Ochs
Feb 10, 2009·Methods in Enzymology·Minoo Razi, Sharon A Tooze
Aug 18, 2009·Journal of Visualized Experiments : JoVE·James Lim, Gaudenz Danuser
Sep 28, 2010·Methods in Cell Biology·Jan R T van WeeringPaul Verkade

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