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Use of an encapsulated fluorescent probe to measure intracellular P O2
Author(s) -
Podgorski Gary T.,
Longmuir Ian S.,
Knopp James A.,
Benson Douglas M.
Publication year - 1981
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.1041070304
Subject(s) - amoeba proteus , nanocapsules , fluorescence , oxygen , laboratory flask , pyrene , chemistry , intracellular , biophysics , quenching (fluorescence) , analytical chemistry (journal) , polyacrylamide , chromatography , nanotechnology , materials science , biochemistry , organic chemistry , optics , biology , polymer chemistry , nanoparticle , physics , cytoplasm
The objectives of this investigation were to produce a reliable, senstive probe to measure intracellular P O2 with a high degree of resolution and to apply this technique to biological systems. A fluorescent molecule, pyrene dissolved in paraffin oil, was encapsulated in polyacrylamide to form a probe of nanometer dimensions. The quantitative and microscopic oxygen values were determined by analyzing the quenching of the fluorescence of the probe by oxygen, as displayed on a television monitor by a silicon‐intensified‐target camera. The nanocapsules had a sensitivity of approximately 1 mm P O2 , a spatial resolution of 0.5 μm, and a temporal resolution of milliseconds. Calibrated nanocapsules within nonrespiring Amoeba proteus responded to ambient partial pressures of oxygen. At two different ambient partial pressures, nanocapsules engulfed by respiring amoebas indicated an intracellular P O2 28 mm Hg less than extracellular P O2 . The capsules retained their sensitivity to oxygen for at least 8 months.