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J Appl Physiol 100: 1301-1310, 2006. First published December 15, 2005; doi:10.1152/japplphysiol.01315.2005
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INNOVATIVE METHODOLOGY

Dual-wavelength phosphorimetry for determination of cortical and subcortical microvascular oxygenation in rat kidney

Tanja Johannes,1,2,* Egbert G. Mik,1,* and Can Ince1

1Department of Physiology, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands; and 2Department of Anesthesiology and Critical Care, University Hospital Tuebingen, Tuebingen, Germany

Submitted 14 October 2005 ; accepted in final form 14 December 2005

This study presents a dual-wavelength phosphorimeter developed to measure microvascular PO2 (µPO2) in different depths in tissue and demonstrates its use in rat kidney. The used phosphorescent dye is Oxyphor G2 with excitation bands at 440 and 632 nm. The broad spectral gap between the excitation bands combined with a relatively low light absorption of 632 nm light by tissue results in a marked difference in penetration depths of both excitation wavelengths. In rat kidney, we determine the catchments depth of the 440-nm excitation to be 700 µm, whereas the catchments depth of 632 nm is as much as 4 mm. Therefore, the measurements differentiate between cortex and outer medulla, respectively. In vitro, no difference in PO2 readings between both channels was found. On the rat kidney in vivo, the measured cortical µPO2 was on average 20 Torr higher than the medullary µPO2 over a wide PO2 range induced by variations in inspired oxygen fraction. Examples provided from endotoxemia and resuscitation show differences in responses of mean cortical and medullary PO2 readings as well as in the shape of the PO2 histograms. It can be concluded that oxygen-dependent quenching of phosphorescence of Oxyphor G2 allows quantitative measurement of µPO2 noninvasively in two different depths in vivo. Oxygen levels measured by this technique in the rat renal cortex and outer medulla are consistent with previously published values detected by Clark-type oxygen electrodes. Dual-wavelength phosphorimetry is excellently suited for monitoring µPO2 changes in two different anatomical layers under pathophysiological conditions with the characteristics of providing oxygen histograms from two depths and having a penetration depth of several millimeters.

phosphorescence quenching; oxygen measurement; renal microvascular oxygenation; oxygen distribution



Address for reprint requests and other correspondence: C. Ince, Dept. of Physiology, Academic Medical Center, Univ. of Amsterdam, Meibergdreef 9, 1105 AZ Amsterdam, The Netherlands (e-mail: c.ince{at}amc.uva.nl)




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