FLOW CYTOMETRIC DETERMINATION OF ABSOLUTE MEMBRANE-POTENTIAL OF CELLS

被引:50
|
作者
KRASZNAI, Z
MARIAN, T
BALKAY, L
EMRI, M
TRON, L
机构
[1] DEBRECEN UNIV MED, SCH MED, CTR POSITRON EMISS TOMOG, H-4012 DEBRECEN, HUNGARY
[2] DEBRECEN UNIV MED, SCH MED, DEPT BIOPHYS, H-4012 DEBRECEN, HUNGARY
基金
匈牙利科学研究基金会;
关键词
FLOW CYTOMETRY; FLUORESCENCE; ABSOLUTE MEMBRANE POTENTIAL;
D O I
10.1016/1011-1344(94)07099-A
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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 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 damp measurements.
引用
收藏
页码:93 / 99
页数:7
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