By Myrtle A. Davis
Univ. of Maryland, Baltimore. Discusses laser scanning cytometry for the research of apoptosis, tissue-based equipment of quantification for stories utilizing animal versions, and useful assays and ELISA ideas. DNLM: Apoptosis--physiology.
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Additional info for Apoptosis Methods in Pharmacology and Toxicology: Approaches to Measurement and Quantification
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. , Reynolds, A. , and Bastiaens, P.
For each measured object, the following parameters are recorded by LSC: 1. Integrated fluorescence intensity within the integration contour, which can be adjusted to a desired width with respect to the threshold contour; 2. ” 3. The area within the integration contour. 4. The perimeter of the contour (in micrometers). 5. Integrated fluorescence intensity within the area of a torus of a desired width defined by the peripheral contour located around (outside) the primary integration contour. , over the rim of cytoplasm of desired width outside the nucleus).
Opening of the pore can be inhibited by a number of compounds including cyclosporin A and bongkrekic acid (24). Low molecular-weight hydrophilic dyes such as calcein are normally excluded from the inner mitochondrial space, but will pass through the opened pore (25). Redistribution of calcein to the inner mitochondrial space, which is blocked by cyclosporin A or other inhibitors of the MPT, appears to represent a true pore-opening event. Bcl 2 family members are thought to exert their pro- or antiapoptoic effects, in part, by interacting with and modulating pore opening (26).
Apoptosis Methods in Pharmacology and Toxicology: Approaches to Measurement and Quantification by Myrtle A. Davis