Flow cytometric determination of absolute membrane potential of cells

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Abstract

Membrane potential measurements using fluorescent membrane potential indicator dyes report on relative changes but usually do not result in an absolute value of the measured parameter. The method developed in this paper is based on the assumption that the negatively charged bis-oxonol distributes across the cytoplasmic membrane according to the Nernst equation. It is further supposed that the fluorescence intensity measured from a given stained cell is a single-value function of the intracellular dye concentration. The protocol suggested incorporates the construction of a calibration curve (fluorescence intensity measured from stained cells vs. extracellular dye concentration). This allows the evaluation of the membrane potential in millivolts using the fluorescence readings of the cells both in the depolarized state and in the state of interest. Good agreement was found between absolute membrane potential data of human peripheral blood lymphocytes by our method and results of parallel patch clamp measurements.

Original languageEnglish
Pages (from-to)93-99
Number of pages7
JournalJournal of Photochemistry and Photobiology, B: Biology
Volume28
Issue number1
DOIs
Publication statusPublished - 1995

Fingerprint

membrane potential
Membrane Potentials
membranes
Membranes
dyes
Coloring Agents
Fluorescence
fluorescence
cells
Dyes
Jurisprudence
clamps
Lymphocytes
Calibration
lymphocytes
cell membranes
Clamping devices
Reading
calibration
Cell Membrane

Keywords

  • Absolute membrane potential
  • Flow cytometry
  • Fluorescence

ASJC Scopus subject areas

  • Biophysics
  • Radiology Nuclear Medicine and imaging
  • Radiation
  • Radiological and Ultrasound Technology
  • Plant Science
  • Bioengineering
  • Physical and Theoretical Chemistry

Cite this

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title = "Flow cytometric determination of absolute membrane potential of cells",
abstract = "Membrane potential measurements using fluorescent membrane potential indicator dyes report on relative changes but usually do not result in an absolute value of the measured parameter. The method developed in this paper is based on the assumption that the negatively charged bis-oxonol distributes across the cytoplasmic membrane according to the Nernst equation. It is further supposed that the fluorescence intensity measured from a given stained cell is a single-value function of the intracellular dye concentration. The protocol suggested incorporates the construction of a calibration curve (fluorescence intensity measured from stained cells vs. extracellular dye concentration). This allows the evaluation of the membrane potential in millivolts using the fluorescence readings of the cells both in the depolarized state and in the state of interest. Good agreement was found between absolute membrane potential data of human peripheral blood lymphocytes by our method and results of parallel patch clamp measurements.",
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author = "Zolt{\'a}n Krasznai and Ter{\'e}z M{\'a}ri{\'a}n and L{\'a}szl{\'o} Balkay and Mikl{\'o}s Emri and Lajos Tr{\'o}n",
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T1 - Flow cytometric determination of absolute membrane potential of cells

AU - Krasznai, Zoltán

AU - Márián, Teréz

AU - Balkay, László

AU - Emri, Miklós

AU - Trón, Lajos

PY - 1995

Y1 - 1995

N2 - Membrane potential measurements using fluorescent membrane potential indicator dyes report on relative changes but usually do not result in an absolute value of the measured parameter. The method developed in this paper is based on the assumption that the negatively charged bis-oxonol distributes across the cytoplasmic membrane according to the Nernst equation. It is further supposed that the fluorescence intensity measured from a given stained cell is a single-value function of the intracellular dye concentration. The protocol suggested incorporates the construction of a calibration curve (fluorescence intensity measured from stained cells vs. extracellular dye concentration). This allows the evaluation of the membrane potential in millivolts using the fluorescence readings of the cells both in the depolarized state and in the state of interest. Good agreement was found between absolute membrane potential data of human peripheral blood lymphocytes by our method and results of parallel patch clamp measurements.

AB - Membrane potential measurements using fluorescent membrane potential indicator dyes report on relative changes but usually do not result in an absolute value of the measured parameter. The method developed in this paper is based on the assumption that the negatively charged bis-oxonol distributes across the cytoplasmic membrane according to the Nernst equation. It is further supposed that the fluorescence intensity measured from a given stained cell is a single-value function of the intracellular dye concentration. The protocol suggested incorporates the construction of a calibration curve (fluorescence intensity measured from stained cells vs. extracellular dye concentration). This allows the evaluation of the membrane potential in millivolts using the fluorescence readings of the cells both in the depolarized state and in the state of interest. Good agreement was found between absolute membrane potential data of human peripheral blood lymphocytes by our method and results of parallel patch clamp measurements.

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