By Myrtle A. Davis
Univ. of Maryland, Baltimore. Discusses laser scanning cytometry for the research of apoptosis, tissue-based tools of quantification for experiences utilizing animal types, and useful assays and ELISA strategies. DNLM: Apoptosis--physiology.
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Additional resources for Apoptosis Methods in Pharmacology and Toxicology: Approaches to Measurement and Quantification
2. , in a drawer) (see Note 10). 3. Aliquot cell suspensions into 12 × 75 mm polypropylene tubes such that for each sample two replicate tubes are obtained. Add 1 µL of each of the 200 µM MitoTracker Green FM and CMXRosamine dye stock solutions to each tube with 1 mL of prewarmed cell suspension. Add 5 µL of monobromobimane stock Apoptotic Cell Death 29 solution to half of the samples. Mix immediately by briefly vortexing at maximal speed. Incubate for 30 min. at 37°C in the dark or at subdued light.
Cytogram of MitoTracker Green FM and CMXRosamine fluorescence (A) and cell-cycle histograms of “normal” (B) and “compromised” (C) cells from a culture of human lymphoblastoid cells that were treated with 1 mg/mL mitomycin C for 8 h at 37°C, harvested and stained according to Protocol 1. Apoptotic cells show a lower CMXRosamine and a higher MitoTracker Green FM fluorescence (A). The two right hand panels show the cell-cycle distributions, based on Hoechst 33342 fluorescence, of the “normal” and “compromised” cells.
75(5), 2547–2557. 78. , and Webb, W. W. (1999) Molecular dynamics in living cells observed by fluorescence correlation spectroscopy with one- and two-photon excitation. Biophys. J. 77(4), 2251–2265. 79. , and Webb, W. W. (1999) Fluorescence correlation spectroscopy with single-molecule sensitivity on cell and model membranes. Cytometry 36(3), 176–182. 80. , et al. (1999) Imaging protein kinase Calpha activation in cells. Science 283(5410), 2085–2089. 36 Poot et al. 81. Verveer, P. , Wouters, F.