Electroporative adjustment of pH in living yeast cells: ratiometric fluorescence pH imaging
Jazyk angličtina Země Nizozemsko Médium print
Typ dokumentu časopisecké články, práce podpořená grantem
- MeSH
- arylsulfonany chemie MeSH
- elektroporace * MeSH
- fluorescenční barviva chemie MeSH
- fluorescenční mikroskopie metody MeSH
- kalibrace MeSH
- koncentrace vodíkových iontů * MeSH
- protony MeSH
- Saccharomyces cerevisiae chemie cytologie metabolismus MeSH
- vodík analýza metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- arylsulfonany MeSH
- fluorescenční barviva MeSH
- protony MeSH
- pyranine MeSH Prohlížeč
- vodík MeSH
A number of vital cell functions including modulation of signaling pathways and regulation of the cellular transport critically depends on the cytoplasmic pH. Many pathological cellular changes are related to the abnormal cytosolic pH as well. Reliable and well-calibrated methods for quantification of the cytosolic pH are therefore of high importance. The pH calibration is particularly difficult in walled cells since standard methods fail. In this report we evaluated the new electroporative calibration method of the cytosolic pH in yeasts by the fluorescence microscopy. The calibration was done on living cells using pyranine as a ratiometric pH-sensitive probe. The probe was electroporatively delivered to the cytosol. We have shown that unlike the measurements in suspension the fluorescence microscopy reveals cell subpopulations with different sensitivity to the pH calibration. While the majority of the cells were well calibrated, there was found subpopulation of uncalibrated cell as well as singular cells exhibiting anomalous pH calibration due to the staining of acidic organelles. Resolution of cell subpopulations helps to achieve better pH calibration compared to the calibration in cuvette on a cell suspension.
Zobrazit více v PubMed
Cytometry A. 2003 Apr;52(2):77-89 PubMed
Res Microbiol. 2000 Jan-Feb;151(1):43-52 PubMed
Crit Rev Biotechnol. 1996;16(4):349-62 PubMed
Yeast. 1998 Apr 30;14(6):501-15 PubMed
Fungal Genet Biol. 2000 Mar;29(2):61-71 PubMed
Biochem Biophys Res Commun. 1999 Aug 2;261(2):377-80 PubMed
Epilepsia. 1992 Sep-Oct;33(5):775-84 PubMed
Photochem Photobiol. 2001 Jul;74(1):8-13 PubMed
Histochem J. 1999 Oct;31(10):635-43 PubMed
Infect Immun. 2000 Mar;68(3):1061-8 PubMed
Bioelectrochemistry. 2003 Oct;61(1-2):65-72 PubMed
Biophys J. 1997 Nov;73(5):2630-7 PubMed
J Exp Med. 1998 May 18;187(10):1583-98 PubMed
Folia Microbiol (Praha). 1996;41(1):98-100 PubMed
Physiol Rev. 1999 Oct;79(4):1089-125 PubMed
Anal Biochem. 1987 Dec;167(2):362-71 PubMed
Planta. 2000 Feb;210(3):347-70 PubMed
Biochim Biophys Acta. 1991 Dec 3;1098(1):79-89 PubMed
Bioelectrochem Bioenerg. 1999 Feb;48(1):3-16 PubMed
Annu Rev Biochem. 1976;45:501-30 PubMed
Cytometry. 1992;13(8):795-808 PubMed
Prog Neurobiol. 1996 Feb;48(2):73-103 PubMed
J Bacteriol. 1995 Feb;177(4):1017-22 PubMed
Biochim Biophys Acta. 1978 May 18;509(2):289-99 PubMed
Biophys J. 2003 Apr;84(4):2709-14 PubMed
Anal Biochem. 2004 Jun 15;329(2):348-50 PubMed
FEBS Lett. 1982 Apr 5;140(1):22-6 PubMed
