Correlative Light Electron Microscopy

In correlative light-electron microscopy (CLEM) technique the fate of GFP-marked membrane structures or tagged proteins can be followed first in living cells and then the fluorescent pattern can be confirmed at the ultra structural level by processing the same cell further for immunolabelling using pre-embedding immunolabelling. 

Image 1 In fluorescence images (left) a dividing cell is progressing from metaphase to anaphase, and then the cell was fixed and processed for immunoEM. Green colour and black precipitate represent GFP-tagged ER-protein and blue colour chromosomes. (M. Puhka).

Image 2 Epifluorescence image (inset in A) and low power TEM image (A) of the same primary sympathetic neuron showing that over-expressed GFP-XIAP is accumulated into the inclusion bodies (bright spots in inset, arrows in A and high-power image of one of the structures in B). Immunolabelling using anti-GFP antibodies verified that the inclusion bodies found in TEM images corresponded to the bright spots imaged with fluorescence microscope (Yu et al., 2003, MCN 22:308-318).

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