High-quality immunofluorescence of cultured cells.
Immunofluorescence microscopy of cultured cells often gives poor preservation of delicate structures. We have obtained dramatically improved results with a simple modification of a standard protocol. Cells growing on a coverslip are rapidly dehydrated in a cold organic solvent and then are rehydrated in a solution containing a homobifunctional crosslinker. The crosslinking reaction stabilizes cellular structures during subsequent incubation and wash steps, usually without compromising antigenicity. This method reproducibly yields high-quality images of endomembrane compartments and cytoskeletal elements.
Bhattacharyya
Dibyendu
D
Department of Molecular Genetics and Cell Biology, The University of Chicago, Chicago, IL, USA.
Hammond
Adam T
AT
Glick
Benjamin S
BS
D000818:Animals; D007668:Kidney / Q000166:cytology; D008856:Microscopy, Fluorescence / Q000379:methods*; D011309:Preservation, Biological; D051381:Rats