Abstract
Fura-2 is the most commonly used member of a family of calcium sensitive flourescent dyes that allows the measurement of intracellular free calcium (Cai by dual excitation fluorimetry. The use of this dye in conjunction with video imaging microscopy permits visualization of changes in Cai with a high degree of spatial resolution. This resolution in turn allows the detection of Cai gradients, waves, and other localized cellular responses not easily detected by photometric techniques. Despite these advantages, most published reports have utilized photometry rather than imaging, due largely to the higher cost and greater complexity of use of the latter technique. With these problems in mind, we have developed a turnkey calcium imaging system that is both inexpensive and simple enough to use that it can be mastered in 2-3 hours.Data showing the distribution of free Ca2+ in the cytoplasm of a quiescent human fibroblast and the kinetics of stimulation of these cells with mitogens are illustrated on the next page.
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