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2′,7′-Bis-(2-carboxyethyl)-5(6)-carboxyfluorescein as a dual-emission fluorescent indicator of intracellular pH suitable for argon laser confocal microscopy

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Abstract

The widely used fluorescent probe 2′,7′-bis-(2-carboxyethyl)-5(6)-carboxyfluorescein (BCECF) serves as a pH-sensitive indicator in classical microscopy. Characteristics of BCECF were studied and a way of employing the probe in a confocal laser scanning microscope equipped with an argon laser at 488 nm was developed, based on the fact that the emission fluorescence spectra are pH-dependent with spectral maximum shift from 518 to 529 nm. Optical filters for the dual-emission ratio method were set to 506 and 529 nm. pH values measured inside a single cell ofSaccharomyces cerevisiae were similar to those obtained with other pH estimation methods.

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Authors and Affiliations

  1. Institute of Physiology, Academy of Sciences of the Czech Republic, 142 20, Prague, Czech Republic

    E. Lanz, J. Slavík & A. Kotyk

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  1. E. Lanz
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  2. J. Slavík
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  3. A. Kotyk
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Lanz, E., Slavík, J. & Kotyk, A. 2′,7′-Bis-(2-carboxyethyl)-5(6)-carboxyfluorescein as a dual-emission fluorescent indicator of intracellular pH suitable for argon laser confocal microscopy. Folia Microbiol 44, 429–434 (1999). https://doi.org/10.1007/BF02903718

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  • DOI: https://doi.org/10.1007/BF02903718

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