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 dye—quin2—is 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 (∼10−7M) and below (to 10−8M), whereas all other techniques do their best at detecting activated levels (∼10−6M) 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 indicators—fura-2 and indo-1—as 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. • Keywords: calcium, quin2 • Bioblast editor: Cecatto C
Labels: MiParea: Instruments;methods
Regulation: Calcium
2020-05, Ca-list
