Enhanced sensitivity of pHluorin-based monitoring of intracellular pH changes achieved through synchronously scanned fluorescence spectra
Language English Country United States Media print-electronic
Document type Journal Article, Research Support, Non-U.S. Gov't
- MeSH
- Spectrometry, Fluorescence MeSH
- Hydrogen-Ion Concentration MeSH
- Saccharomyces cerevisiae chemistry cytology metabolism MeSH
- Green Fluorescent Proteins chemistry MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- PHluorin MeSH Browser
- Green Fluorescent Proteins MeSH
Since its introduction in 1998, genetically encoded pH-sensitive sensor ratiometric pHluorin proved to be a valuable tool for cell physiology studies. Here, we show how the sensitivity of pHluorin-based monitoring of intracellular pH changes performed with cell suspensions can be enhanced by using synchronously scanned fluorescence spectroscopy. In the suspensions of S. cerevisiae cells subjected to varying extracellular pH values, we have been able to measure statistically significant changes in intracellular pH of less than 0.1 unit, which were not detectable using a standard ratiometric approach.
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H+ translocation by weak acid uncouplers is independent of H+ electrochemical gradient
