We describe a new method for calibrating intracellular pH (pH,) measurements by flow cytometry, based on the null point method proposed originally by Eisner et al. (Pflugers Arch 413:55+558, 1989). The method involves suspending cells loaded with pH-sensitive dyes, such as SNARF-1 or BCECF, in deh
Measurement of fast dynamic intracellular pH in Saccharomyces cerevisiae using benzoic acid pulse
โ Scribed by M.T.A.P. Kresnowati; C. Suarez-Mendez; M.K. Groothuizen; W.A. van Winden; J.J. Heijnen
- Book ID
- 101722834
- Publisher
- John Wiley and Sons
- Year
- 2007
- Tongue
- English
- Weight
- 255 KB
- Volume
- 97
- Category
- Article
- ISSN
- 0006-3592
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โฆ Synopsis
Abstract
pH affects many processes on cell metabolism, such as enzyme kinetics. To enhance the understanding of the living cells, it is therefore indispensable to have a method to monitor the pH in living cells. To accomplish this, a dynamic intracellular pH measurement method applying low concentration benzoic acid pulse was developed. The method was thoroughly validated and successfully implemented for measuring fast dynamic intracellular pH of Saccharomyces cerevisiae in response to a glucose pulse perturbation performed in the BioSCOPE setโup. Fast drop in intracellular pH followed by partial alkalinization was observed following the pulse. The low concentration benzoic acid pulse which was implemented in the method avoids the undesirable effects that may be introduced by benzoic acid to cell metabolism. Biotechnol. Bioeng. 2007;97:86โ98. ยฉ 2006 Wiley Periodicals, Inc.
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