Process for determining the pH value in the interior of a cell

Details Process for determining the pH value in the interior of a cell Process for determining the pH value in the interior of a cell

1982-02-05

1985-03-05

Kepplinger, Esther M.

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving viable micro-organism

435 4, 435 19, 435 23, C12Q 102, C12Q 100, C12Q 138, C12Q 144

Patent

active

045031487

ABSTRACT:
A process for determining the pH value in the interior of a cell is described. This is accomplished by measurement of the emission of a fluorescent substance which is absorbed by the cell through incubation in a solution. The solution contains a compound of the said fluorescent substance; the fluorescent substance is separated from this compound in the cell by an enzyme present in the cell. The fluorescent substance selected is one such that the frequency of the emission maximum is dependent on the pH value. The compound of this fluorescent substance which is selected is one which is membrane penetrating. Measurement of the fluorescence is based on the principle of measurement of the frequency at which the emission maximum occurs. A fluorescent substance suitable for this purpose is 2,3-dicyano-1,4-hydroquinone. A suitable membrane penetrating compound is 1,4-diacetoxy-2,3-dicyanobenzene or 1,4-dibutyryloxy-2,3-dicyanobenzene or 1,4-di(-tert-butyloxycarbonyl-1-alanyloxy)-2,3-dicyanobenzene.

REFERENCES:
Visser et al., J. Histochem. Cytochem., 27(1): 32-35, (1979).
Thomas et al., Biochemistry, 18(11): 2210-2218, (1979).
Visser et al., Chemical Abstracts, 90: 148002f, 258, (1979).

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