BCECF2',7'-Bis- (2-Carboxyethyl)-5- (And-6)- carboxyfluorescein (molecular probes)
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BCECF is a neutral lipophilic form of bis-carboxyfluorescein which diffuses freely through the plasma membrane.
The fluorescence intensity of BCECF is dependent upon the pH with a maximum response at an excitation of [lambda]=535 nm (F1), while at [lambda]=490 nm (F2), the intensity is independent of pH.
To determine pHi, a non-carbonate solution and BCECF, an intracellular fluorescent dye, were used and the light density was found using a spectroflourometer.