A Versatile Method for Immunofluorescent Staining of Cells Cultured on Permeable Membrane Inserts.
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.
Department of Obstetrics & Gynecology, University of Texas Health Sciences Center at Houston, Houston, TX, USA.
Jenni L
JL
Gillespie
Department of Medical Education - Otolaryngology, Michigan State University College of Medicine Statewide Campus System, Metro Health Hospital, Wyoming, MI, USA.
Anwuli
A
Anyah
Department of Pharmacology and Microbiology, Kansas City University of Medicine and Biosciences, Kansas City, MO, USA.
John M
JM
Taylor
Department of Pharmacology and Microbiology, Kansas City University of Medicine and Biosciences, Kansas City, MO, USA.
Jerry W
JW
Marlin
Department of Biomedical Sciences, Oakland University William Beaumont School of Medicine, Rochester, MI, USA.
Tracey A H
TA
Taylor
D002463:Cell Membrane Permeability; D005455:Fluorescent Antibody Technique / Q000379:methods*; D006367:HeLa Cells; D006801:Humans; D008567:Membranes, Artificial; D008856:Microscopy, Fluorescence / Q000379:methods; D013194:Staining and Labeling / Q000379:methods
D008567:Membranes, Artificial