Ratiometric analysis of calcium mobilization
Clinical and Applied Immunology Reviews(2000)
摘要
One of the earliest events in lymphocyte activation is an increase in the concentration of cytosolic free Ca2+. Antigen recognition, chemotaxis, apoptosis and cell cycle progression are all associated with changes in cytosolic free Ca2+ levels. The kinetics, subcellular localization, and degree of Ca2+ mobilization can be assessed with fluorescent Ca2+ indicators. Ratiometric analysis of Ca2+ mobilization reduces errors due to variations in cell size and shape, indicator loading and instrument variability that can otherwise limit the usefulness of fluorescent Ca2+ indicators. The purposes of this paper are to summarize key characteristics of some useful Ca2+-sensitive fluorescent dyes (Indo-1, Fura-2, Fluo-3, Fluo-4 and Fura Red) and to review published methods for the optimization of Ca2+ mobilization analysis with ratiometric indicators. Analysis of Ca2+ mobilization provides a rapid, quantitative readout of lymphocyte activation.
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关键词
AM, acetoxymethyl,BAPTA, 1,2-bis(2-aminophenoxy)ethane-N, N, N′, N′-tetraacetic acid,[Ca2+]cyto, cytoplasmic ionized calcium concentration,DMSO, dimethyl sulfoxide,EGTA, ethylene glycol bis(β-aminoethyl ether)-N, N, N′, N′-tetraacetic acid,IP3, inositol 1,4,5 trisphosphate
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