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Proceedings Paper

Confocal fluorescence lifetime imaging of pH in single cells
Author(s): Renata Sanders; Hans C. Gerritsen; Arie Draaijer; P. M. Houpt; Sjaak J. F. van Veen; Yehudi K. Levine
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Paper Abstract

We show that the confocal fluorescence lifetime imaging method is a powerful tool for the quantitative determination of pH on a microscopic scale. This method is easily implemented using a conventional confocal microscope and moreover, utilizes currently available fluorescent probes. It is shown that both the intensity probe DM-NERF and the ratio probe BCECF are suitable for this purpose, albeit with different useful pH ranges. In addition it is shown that the fluorescence decay of both probes are independent of the probe-concentration. Furthermore, the fluorescence lifetime behavior of BCECF is found to be insensitive to changes in the hydrophobicity and protein content of the buffer solution. The intracellular pH was imaged using BCECF since this probe is sensitive in the physiological pH range. A realistic pH value of about 7.3 was found throughout CHO cells.

Paper Details

Date Published: 17 August 1994
PDF: 7 pages
Proc. SPIE 2137, Time-Resolved Laser Spectroscopy in Biochemistry IV, (17 August 1994); doi: 10.1117/12.182761
Show Author Affiliations
Renata Sanders, Univ. of Utrecht and TNO Institute of Environmental Sciences (Netherlands)
Hans C. Gerritsen, Univ. of Utrecht (Netherlands)
Arie Draaijer, TNO Institute of Environmental Sciences (Netherlands)
P. M. Houpt, TNO Institute of Environmental Sciences (Netherlands)
Sjaak J. F. van Veen, TNO Institute of Environmental Sciences (Netherlands)
Yehudi K. Levine, Univ. of Utrecht (Netherlands)

Published in SPIE Proceedings Vol. 2137:
Time-Resolved Laser Spectroscopy in Biochemistry IV
Joseph R. Lakowicz, Editor(s)

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