Rapid spectrophotometric determination of oxygen consumption using hemoglobin, in vitro: Light scatter correction and expanded dynamic range

Paul R. Territo, Robert S. Balaban

Research output: Contribution to journalArticle

12 Scopus citations

Abstract

The method of using absorbance in conjunction with hemoglobin (Hb) to monitor rapid changes in oxygen consumption in vitro was improved by using a nonlinear calibration technique and multiwavelength spectroscopy. The O2 dependence of Hb absorbance was effectively linearized using the current technique (R2 = 0.990 ± 0.002, n = 3), and extended the dynamic range of [O2] determinations by 1.6-fold over previous approaches. The association/dissociation rates of O2 and Hb were evaluated using the current approach and were not significant on the 100-ms time domain. A method was also developed for compensating for large amplitude light scattering changes in turbid media using multiwavelength analysis. Both the nonlinear calibration curve and light scattering corrections were validated in isolated porcine heart mitochondrial preparations. (C) 2000 Academic Press.

Original languageEnglish (US)
Pages (from-to)156-163
Number of pages8
JournalAnalytical biochemistry
Volume286
Issue number1
DOIs
StatePublished - Nov 1 2000

Keywords

  • Hemoglobin
  • Light scattering
  • Metabolism
  • Response time
  • Spectrophotometric

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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