title: Method for collecting and immobilizing individual cumulus cells enabling quantitative immunofluorescence analysis of proteins.
abstract: Most immunofluorescence methods rely on techniques dealing with a very large number of cells. However, when the number of cells in a sample is low (e.g., when cumulus cells must be analyzed from individual cumulus-oocyte complexes), specific techniques are required to conserve, fix, and analyze cells individually. We established and validated a simple and effective method for collecting and immobilizing low numbers of cumulus cells that enables easy and quick quantitative immunofluorescence analysis of proteins from individual cells. To illustrate this technique, we stained proprotein of a disintegrin and metalloproteinase with thrombospondin-like repeats-1 (proADAMTS-1) and analyzed its levels in individual porcine cumulus cells.
authors: Department of Reproduction, Obstetrics, and Herd Health, Faculty of Veterinary Medicine, Ghent University, B-9820 Merelbeke, Belgium. Electronic address: ruth.appeltant@ugent.be.
authors: R
authors: R
authors: Appeltant
authors: Department of Reproduction, Obstetrics, and Herd Health, Faculty of Veterinary Medicine, Ghent University, B-9820 Merelbeke, Belgium.
authors: D
authors: D
authors: Maes
authors: Department of Reproduction, Obstetrics, and Herd Health, Faculty of Veterinary Medicine, Ghent University, B-9820 Merelbeke, Belgium.
authors: A
authors: A
authors: Van Soom
mesh_terms: D000818:Animals; D054885:Cumulus Cells / Q000737:chemistry*; D019483:Disintegrins / Q000032:analysis*; D005455:Fluorescent Antibody Technique; D020782:Matrix Metalloproteinases / Q000032:analysis* / Q000378:metabolism; D013552:Swine
chemical_list: D019483:Disintegrins; D020782:Matrix Metalloproteinases
keywords: Cell-Tak; Cumulus cells; Immobilization; Protein immunostaining; proADAMTS-1