Comparison of DNA Fragmentation and Color Thresholding for Objective Quantitation of Apoptotic Cells

Authors

D. R. Plymale, Tulane University
D. S. Ng Tang, Baylor College of Medicine
C. D. Fermin, Tulane University
D. E. Lewis, Baylor College of Medicine
D. S. Martin, Tulane University
R. F. Garry, Tulane UniversityFollow

Abstract

Apoptosis is a process of cell death characterized by distinctive morphological changes and fragmentation of cellular DNA. Using video imaging and color thresholding techniques, we objectively quantitated the number of cultured CD4 + T-lymphoblastoid cells (HUT78 cells, RH9 subclone) displaying morphological signs of apoptosis before and after exposure to γ-irradiation. The numbers of apoptotic cells measured by objective video imaging techniques were compared to numbers of apoptotic cells measured in the same samples by sensitive apoptotic assays that quantitate DNA fragmentation. DNA fragmentation assays gave consistently higher values compared with the video imaging assays that measured morphological changes associated with apoptosis. These results suggest that substantial DNA frag-mentation can precede or occur in the absence of the morphological changes which are associated with apoptosis in γ-irradiated RH9 cells.

Recommended Citation

Plymale, D. R.; Ng Tang, D. S.; Fermin, C. D.; Lewis, D. E.; Martin, D. S.; and Garry, R. F. (1995) "Comparison of DNA Fragmentation and Color Thresholding for Objective Quantitation of Apoptotic Cells," Scanning Microscopy: Vol. 9: No. 3, Article 20.
Available at: https://digitalcommons.usu.edu/microscopy/vol9/iss3/20