2006
DOI: 10.1529/biophysj.105.069450
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Simultaneous Measurement of Water Volume and pH in Single Cells Using BCECF and Fluorescence Imaging Microscopy

Abstract: Regulation and maintenance of cell water volume and intracellular pH (pHi) are vital functions that are interdependent; cell volume regulation affects, and is in turn affected by, changes in pHi. Disruption of either function underlies various pathologies. To study the interaction and kinetics of these two mechanisms, we developed and validated a quantitative fluorescence imaging microscopy method to measure simultaneous changes in pHi and volume in single cells loaded with the fluorescent probe BCECF. CWV is … Show more

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Cited by 34 publications
(38 citation statements)
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“…In this way, relative volume changes can be determined. 28 This is illustrated in Figure 3A for a representative D54-GFP cell prior to (left), during (center), and after M Phase (right). The fluorescent images demonstrate an increase in the intensity of GFP at M phase which corresponds to a concentration of the protein and hence a decrease in cell volume.…”
Section: Resultsmentioning
confidence: 98%
“…In this way, relative volume changes can be determined. 28 This is illustrated in Figure 3A for a representative D54-GFP cell prior to (left), during (center), and after M Phase (right). The fluorescent images demonstrate an increase in the intensity of GFP at M phase which corresponds to a concentration of the protein and hence a decrease in cell volume.…”
Section: Resultsmentioning
confidence: 98%
“…In the case of fluorescent molecules, changes in concentration are reflected in changes in fluorescence intensity (Alvarez- Leefmans et al 2006). We use the soluble GFP expressed by the 1-D54 cells to measure intensity over time in the presence or absence of GABA.…”
Section: R E S U L T Smentioning
confidence: 99%
“…The fluorescence of the cell suspension was measured using spectrofluorimeter (Aminco Bowman series 2, SLM-Aminco, Edison, NJ). The signal was plotted as the ratio of the intensities at 535 nm with excitation at 440 and 490 nm [31].…”
Section: Measurement Of Intracellular Ph Changementioning
confidence: 99%