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In situ calibrations for BCECF within Arabidopsis root columella cells showed a pH-dependent increase in ratio when the fluorescent emission using 490 nm of excitation (pH-dependent) was ratioed against emission with 440 nm excitation. Reliable pHc values may be obtained with BCECF between 5.5 and 8.0, since the pKa of BCECF was estimated at 6.8, which is close to the published value of 6.98.
