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A Versatile Method for Immunofluorescent Staining of Cells Cultured on Permeable Membrane Inserts

Jenni L. Gillespie, Anwuli Anyah, John M. Taylor, Jerry W. Marlin, Tracey A.H. Taylor

Department of Obstetrics & Gynecology, University of Texas Health Sciences Center at Houston, Houston, TX, TX, USA

Med Sci Monit Basic Res 2016; 22:91-94

DOI: 10.12659/MSMBR.900656

Available online: 2016-09-12

Published: 2016-09-12


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