Enhanced sensitivity of pHluorin-based monitoring of intracellular pH changes achieved through synchronously scanned fluorescence spectra.
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.
Charles University , Faculty of Mathematics and Physics, Ke Karlovu 3, 12116 Prague, Czech Republic.
Jaromír
J
Plášek
Charles University , Faculty of Mathematics and Physics, Ke Karlovu 3, 12116 Prague, Czech Republic.
Adéla
A
Melcrová
Charles University , Faculty of Mathematics and Physics, Ke Karlovu 3, 12116 Prague, Czech Republic.
Dana
D
Gášková
D049452:Green Fluorescent Proteins / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D012441:Saccharomyces cerevisiae / Q000737:chemistry* / Q000166:cytology* / Q000378:metabolism; D013050:Spectrometry, Fluorescence
C485187:PHluorin; D049452:Green Fluorescent Proteins