Improving Membrane Staining of Cultured Cells Using Ferrocyanide as a Post-Fixative

Lack of contrast is a common problem encountered when doing TEM of cultured cells, especially of membranes. Using ferrocyanide as a post-fixative can greatly improve membrane fixation and staining. This protocol has been used to study Caco-2 cells grown on PET membranes (the “Caco model”). Caco-2 is...

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Bibliographic Details
Published in:Microscopy today 2007-03, Vol.15 (2), p.38-39
Main Author: Nizet, Stéphane
Format: Magazinearticle
Language:English
Online Access:Get full text
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Summary:Lack of contrast is a common problem encountered when doing TEM of cultured cells, especially of membranes. Using ferrocyanide as a post-fixative can greatly improve membrane fixation and staining. This protocol has been used to study Caco-2 cells grown on PET membranes (the “Caco model”). Caco-2 is a colon cancer cell line that differentiates upon reaching confluency. This allows permeability studies on a cell model, which is reasonably similar to the human intestine. Basically, the protocol is classical, the only peculiarity consisting in including ferrocyanide in post-fixation. I describe how I prepare and embed the membrane in order to obtain transverse sections of a cell monolayer because I find this is the only way to obtain regular sections with the cells sticking to the membrane (otherwise the ultrathin section splits between the cell and membrane).
ISSN:1551-9295
2150-3583
DOI:10.1017/S1551929500051014