Phase fluorometric sterilizable optical oxygen sensor

We report here on a low‐cost, optical oxygen sensor as an attractive alternative to the widely used amperometric Clark‐type oxygen electrode for measuring dissolved oxygen tensions in cell cultures and bioreactor. Our sensor is based on the defferential quenching of the fluorescence lifetime of chromophore in response to the partial pressure of oxygen. This is measured as a phase shift in fluorescence emission from the chromophore due to oxygen quenching when excited by an intensity modulated beam of light. In this article we demonstrate the advantages of lifetime‐based optical methods over both intensity based optical methods and amperometric electrodes. Our sensor is particularly suitable for measuring dissolved oxygen in bioreactors. It is autoclavable, is free of maintenance requirements, and solvents the problems of long‐term stability, calibration drifts, and reliable measurement of low oxygen tensions in dense microbial cultures that limit the utility of Clark‐type elcectordes. © 1994 John Wiley & Sons, Inc.

[1]  Otto S. Wolfbeis,et al.  Long-wavelength fluorescent indicators for the determination of oxygen partial pressures , 1984 .

[2]  Benjamin A. DeGraff,et al.  Photophysics and photochemistry of oxygen sensors based on luminescent transition-metal complexes , 1991 .

[3]  Max E. Lippitsch,et al.  Fibre-optic oxygen sensor with the fluorescence decay time as the information carrier , 1988 .

[4]  Joseph R. Lakowicz,et al.  Optical sensing of glucose using phase-modulation fluorimetry , 1993 .

[5]  D. Lübbers,et al.  Theory and Development of Fluorescence-Based Optochemical Oxygen Sensors: Oxygen Optodes , 1987, International anesthesiology clinics.

[6]  D. Webb,et al.  Photoluminescence of solutions , 1969 .

[7]  J. Lakowicz.  Principles of fluorescence spectroscopy , 1983 .

[8]  J. Lakowicz,et al.  CALCIUM‐DEPENDENT FLUORESCENCE LIFETIMES OF INDO‐1 FOR ONE‐ AND TWO‐PHOTON EXCITATION OF FLUORESCENCE , 1993, Photochemistry and photobiology.

[9]  Joseph R. Lakowicz,et al.  Fluorescence lifetime-based sensing of pH, Ca2+, K+ and glucose , 1993 .

[10]  James N. Demas,et al.  Determination of oxygen concentrations by luminescence quenching of a polymer-immobilized transition-metal complex , 1987 .

[11]  M. Grätzel,et al.  Synthesis and photophysical characterization of highly luminescent complexes of Ru(II) containing 4,4′-di-(p-carboxyphenyl)-2,2′-bipyridine , 1992 .

[12]  Benjamin A. DeGraff,et al.  Design and Applications of Highly Luminescent Transition Metal Complexes , 1991 .

[13]  W. Baeyens,et al.  Luminescence Techniques in Chemical and Biochemical Analysis , 1990 .

[14]  J. Lakowicz,et al.  Electroluminescent lamp-based phase fluorometer and oxygen sensor. , 1992, Analytical biochemistry.

[15]  H Szmacinski,et al.  Fluorescence lifetime imaging of calcium using Quin-2. , 1992, Cell calcium.

[16]  George T. Tsao,et al.  Dissolved oxygen electrodes , 1979 .