12 September 2016 : Short communication
A Versatile Method for Immunofluorescent Staining of Cells Cultured on Permeable Membrane Inserts
Jenni L. GillespieABCDE, Anwuli AnyahBCDEF, John M. TaylorBCDE, Jerry W. MarlinBE, Tracey A.H. TaylorABCDEFGDOI: 10.12659/MSMBR.900656
Med Sci Monit Basic Res 2016; 22:91-94
Abstract
BACKGROUND
Obtaining high-quality images of cellular structures via immunofluorescence staining is critical for cellular localization studies. Often, these studies cannot be performed in parallel with certain oncology, virology, pharmacokinetic, and drug absorption studies due to model system technicalities requiring the cells to be cultured on porous membranes rather than glass or plastic.
MATERIAL AND METHODS
Here, we report a method of immunofluorescent staining of cells cultured on permeable membranes.
RESULTS
As proof of principle, HeLa cells grown on Transwell® membrane supports were stained with fluorescently labeled antibodies using this modified immunofluorescence staining method and visualized by fluorescent microscopy.
CONCLUSIONS
This protocol is a convenient alternative to staining cells on glass coverslips, thereby expanding the scope and applications of this important research tool.
Keywords: Diffusion Chambers, Culture, Fluorescent Antibody Technique, Direct, Staining and Labeling
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