A novel method for measuring intracellular pH

Effect of neutron irradiation on pHi of transformed cells

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10 Citations (Scopus)

Abstract

A new flow cytometric method was developed for measuring the intracellular pH (pHi) of mammalian cells using a fluorescent pH indicator dye 2′,7-bis-(2-carboxyethyl)-5(and -6)-carboxyfluorescein (BCECF). Emission intensities (or their ratios) measured from BCECF-loaded cells can be converted into absolute pHi values using appropriate calibration curves. By comparison of several possible measuring and data evaluation procedures a double-ratio method was suggested as the most advantageous protocol to yield reliable intracellular pH data. This method allows pHi to be determined on a cell-by-cell basis corrected for cell volume and change in geometry of input-output optics of the flow cytometer. Our method applies a standard calibration curve and does not necessitate its reconstruction for each new set of measurements. Cells of the OKT-4 and OKT-8 hybridoma lines were exposed to neutron irradiation of different doses. Irradiated cells underwent a biphasic alkalinization; an instantaneous effect detected within 1.5 h was found to be intensified over 24 h. For the interpretation of data we suggest that the increase in cytoplasmic pH following neutron treatment is evoked by two mechanisms.

Original languageEnglish
Pages (from-to)367-375
Number of pages9
JournalJournal of Photochemistry and Photobiology, B: Biology
Volume16
Issue number3-4
DOIs
Publication statusPublished - 1992

Fingerprint

pH effects
Neutron irradiation
Neutrons
neutron irradiation
irradiation
Cells
Calibration
cells
Dosimetry
Optics
Dyes
Geometry
methodology
Optic Flow
calibration
curves
Hybridomas
alkalinization
emissions factor
hybridomas

Keywords

  • Flow cytometry
  • fluorescence
  • intracellular pH
  • ionizing radiation
  • neutron irradiation.
  • transformed cells

ASJC Scopus subject areas

  • Plant Science
  • Bioengineering
  • Physical and Theoretical Chemistry

Cite this

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title = "A novel method for measuring intracellular pH: Effect of neutron irradiation on pHi of transformed cells",
abstract = "A new flow cytometric method was developed for measuring the intracellular pH (pHi) of mammalian cells using a fluorescent pH indicator dye 2′,7-bis-(2-carboxyethyl)-5(and -6)-carboxyfluorescein (BCECF). Emission intensities (or their ratios) measured from BCECF-loaded cells can be converted into absolute pHi values using appropriate calibration curves. By comparison of several possible measuring and data evaluation procedures a double-ratio method was suggested as the most advantageous protocol to yield reliable intracellular pH data. This method allows pHi to be determined on a cell-by-cell basis corrected for cell volume and change in geometry of input-output optics of the flow cytometer. Our method applies a standard calibration curve and does not necessitate its reconstruction for each new set of measurements. Cells of the OKT-4 and OKT-8 hybridoma lines were exposed to neutron irradiation of different doses. Irradiated cells underwent a biphasic alkalinization; an instantaneous effect detected within 1.5 h was found to be intensified over 24 h. For the interpretation of data we suggest that the increase in cytoplasmic pH following neutron treatment is evoked by two mechanisms.",
keywords = "Flow cytometry, fluorescence, intracellular pH, ionizing radiation, neutron irradiation., transformed cells",
author = "L. Balkay and T. M{\'a}ri{\'a}n and M. Emri and L. Tr{\'o}n",
year = "1992",
doi = "10.1016/1011-1344(92)80024-P",
language = "English",
volume = "16",
pages = "367--375",
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TY - JOUR

T1 - A novel method for measuring intracellular pH

T2 - Effect of neutron irradiation on pHi of transformed cells

AU - Balkay, L.

AU - Márián, T.

AU - Emri, M.

AU - Trón, L.

PY - 1992

Y1 - 1992

N2 - A new flow cytometric method was developed for measuring the intracellular pH (pHi) of mammalian cells using a fluorescent pH indicator dye 2′,7-bis-(2-carboxyethyl)-5(and -6)-carboxyfluorescein (BCECF). Emission intensities (or their ratios) measured from BCECF-loaded cells can be converted into absolute pHi values using appropriate calibration curves. By comparison of several possible measuring and data evaluation procedures a double-ratio method was suggested as the most advantageous protocol to yield reliable intracellular pH data. This method allows pHi to be determined on a cell-by-cell basis corrected for cell volume and change in geometry of input-output optics of the flow cytometer. Our method applies a standard calibration curve and does not necessitate its reconstruction for each new set of measurements. Cells of the OKT-4 and OKT-8 hybridoma lines were exposed to neutron irradiation of different doses. Irradiated cells underwent a biphasic alkalinization; an instantaneous effect detected within 1.5 h was found to be intensified over 24 h. For the interpretation of data we suggest that the increase in cytoplasmic pH following neutron treatment is evoked by two mechanisms.

AB - A new flow cytometric method was developed for measuring the intracellular pH (pHi) of mammalian cells using a fluorescent pH indicator dye 2′,7-bis-(2-carboxyethyl)-5(and -6)-carboxyfluorescein (BCECF). Emission intensities (or their ratios) measured from BCECF-loaded cells can be converted into absolute pHi values using appropriate calibration curves. By comparison of several possible measuring and data evaluation procedures a double-ratio method was suggested as the most advantageous protocol to yield reliable intracellular pH data. This method allows pHi to be determined on a cell-by-cell basis corrected for cell volume and change in geometry of input-output optics of the flow cytometer. Our method applies a standard calibration curve and does not necessitate its reconstruction for each new set of measurements. Cells of the OKT-4 and OKT-8 hybridoma lines were exposed to neutron irradiation of different doses. Irradiated cells underwent a biphasic alkalinization; an instantaneous effect detected within 1.5 h was found to be intensified over 24 h. For the interpretation of data we suggest that the increase in cytoplasmic pH following neutron treatment is evoked by two mechanisms.

KW - Flow cytometry

KW - fluorescence

KW - intracellular pH

KW - ionizing radiation

KW - neutron irradiation.

KW - transformed cells

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