Simultaneous In Situ Detection of Apoptosis and Necrosis in Monolayer Cultures by TUNEL and Trypan Blue Staining. (June 1997)

• Record Type: Journal Article
• Authors: Perry, Seth W.; Epstein, Leon G.; Gelbard, Harris A.
• Is Part Of: Biotechniques. Volume 22:Number 6(1997)
• Page Start: 1102
• Page End: 1106
• Publication Date: 1997-06
• Subjects: Biology, Experimental -- Periodicals; Molecular biology -- Periodicals; Medical technology -- Periodicals; Biology, Experimental; Medical technology; Molecular biology; Clinical Laboratory Techniques -- Periodicals; Research -- Periodicals; Medical Laboratory Science -- Periodicals; Periodicals; Electronic journals; 570
• DOI: 10.2144/97226st01
• Languages: English
• ISSNs: 0736-6205

Abstract

A method for simultaneously detecting membrane permeability (characteristic of necrosis) and DNA fragmentation (characteristic of apoptosis) is described. By combining a common dye-exclusion method (Trypan Blue) with a commercially available terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end-labeling (TUNEL) labeling kit, we have succeeded in developing a novel methodology for obtaining permanently mounted slides of monolayer cell cultures double-labeled for DNA fragmentation and cell lysis. This method should facilitate in situ studies of cell death by allowing for a more accurate quantification of total toxicity in monolayer cell cultures and perhaps further enhance our understanding of the different mechanisms of cell death as well.