Correlative Light and Electron Microscopy (CLEM)

Method Specification

Many biological insights can be obtained by combining the power of Fluorescence Microscopy with that of Electron Microscopy to study the same sample – this is called Correlative Light and Electron Microscopy (CLEM).

In Fluorescence Microscopy, specific biological entities can be labelled and identified. In Electron Microscopy, the biological context can be observed in high-resolution. CLEM involves processing specimens for light microscopy and imaging by fluorescence followed by preparation and imaging by electron microscopy.

 

CLEM of intracellular bacteria in human breast cancer. Fluorescence image shows nucleus in blue and bacteria labeling in red. 
[1] Nejman D, Livyatan I, Fuks G, et al. The human tumor microbiome is composed of tumor type-specific intracellular bacteria. Science. 368(6494):973-980 (2020).

 

CLEM of SEM with super resolution STORM of C. clariflavum.
(A, B) cellobiose-grown cells were immunolabeled with anti-GH48 (green) and anti-CohC (pink). (A) Scale bars: 1 μm, 200 nm in each inset. (B) Scale bars: 2 μm, 200 nm in each inset. (C, D) A WS-grown cell was immunolabeled with anti- 21 CohA and anti-CohB. (C) Labeling is colored according to the specific probes: anti-CohA, red; anti-CohB, blue. (D) The depth of labeling is presented for the two combined probes, color-coded from yellow (shallow labeling) to dark red (deep labeling, closer to the slide). Scale bars: 1 μm, 200 nm in each inset.
[1] L. Artzi, T. Dadosh, E. Milrot, S. Morais, S. Levin-Zaidman, E. Morag, E. A. Bayer, Colocalization and Disposition of Cellulosomes in Clostridium clariflavum as Revealed by Correlative Superresolution Imaging. mBio 9, 12 (2018).

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