Monitoring membrane rafts in colorectal cancer cells by means of correlative fluorescence electron microscopy (CFEM).
Detergent-resistant membrane (DRM) rafts have been shown to play a pivotal role in regulating key cell biological processes, such as signal transduction, cellular transport and cell survival. The fine structure of membrane rafts are studied using various different imaging approaches and the outcomes are largely dependent on the detection methodology applied. All these microscopy techniques which employ light-, laser- and photon-optics, electrons as well as atomic force probing are characterized on their turn by their strengths and limitations for membrane raft identification. This explains in part the diversity of definitions available to describe these peculiar membrane structures. We present herewith an alternative and uncomplicated microscopy tool to study fluorescently labelled DRMs with information at the transmission electron microscopical level of the same cell, enabling us to obtain a snapshot of the morpho-functional relationships between the cell's interior and DRMs. The proposed approach of correlative fluorescence electron microscopy (CFEM) can therefore be considered as an additional alternative imaging approach to unravel DRM structure-function relationships from micro- to nanometre length scales, from the cell to the molecule.
Jahn
Kristina A
KA
Australian Key Centre for Microscopy and Microanalysis, Madsen Building F09, The University of Sydney, NSW 2006, Australia.
Braet
Filip
F
D018938:Caco-2 Cells; D015179:Colorectal Neoplasms / Q000648:ultrastructure*; D006801:Humans; D021962:Membrane Microdomains / Q000648:ultrastructure*; D046529:Microscopy, Electron, Transmission; D008856:Microscopy, Fluorescence