Tsien 1989 Methods Enzymol
Tsien R, Pozzan T (1989) Measurement of cytosolic free Ca2+ with quin2. Methods Enzymol 172:230-62. |
Tsien R, Pozzan T (1989) Methods Enzymol
Abstract: The fluorescent Ca2+ indicator dyeβquin2βis the basis of one of the most popular methods for measuring cytosolic free Ca2+ [(Ca2+]i], especially in suspensions of mammalian cells. Quin2 has several advantages over other methods for measuring cytosolic free Ca2+: (1) it can be loaded into the cytosol of millions of cells simultaneously without any micromanipulations or any disruption of the plasma membrane, (2) it is monitored with a conventional cuvette spectrofluorimeter, (3) its readings are usually easily calibrated at typical resting levels of [Ca2+]i (βΌ10β7M) and below (to 10β8M), whereas all other techniques do their best at detecting activated levels (βΌ10β6M) and are nearly at their detection limits at resting levels, and (4) it avoids many of the drawbacks particular to other [Ca2+]i measurement techniques. The chapter compares quin2 with newer fluorescent indicatorsβfura-2 and indo-1βas they are much brighter in fluorescence than quin2. Because fura-2 and indo-1 are sensitive enough to give large calibratable signals from single cells or regions of cells, they allow a direct measurement of compartmentalization and heterogeneity. β’ Keywords: calcium, quin2 β’ Bioblast editor: Cecatto C
Labels: MiParea: Instruments;methods
Regulation: Calcium
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