Measurement of Oxygen in the Microcirculation Using Phosphorescence Quenching Microscopy

  • Roland N. Pittman
  • Aleksander S. Golub
  • Helena Carvalho
Conference paper
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 662)


According to the classical concept of Krogh, O2 is delivered to the tissues solely by capillaries and intra-capillary resistance to O2 diffusion is negligible. Over the past three decades longitudinal PO2 and SO2 gradients in arterioles have been observed with a transmural PO2 gradient in small arterioles of only 1–2 mmHg. Application of phosphorescence quenching microscopy to measurements of PO2 in arterioles of the rat mesentery by Tsai et al. (1998) found a large transmural PO2 in these arterioles. That led to the provocative conclusion that the arteriolar wall is the major sink for O2 in the microcirculation. Our studies indicate that many of these results can be explained by photo-activated O2 consumption following phosphor excitation, combined with a large excitation area and high frequency of flash excitation. We have developed the basic principles for phosphorescence quenching microscopy including the need to use a small excitation area, a low excitation frequency and a scanning excitation for stationary samples.


Excitation Region Hamster Cheek Pouch Arteriolar Wall Flash Frequency Multiple Excitation 
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Supported by grant AHA-0655449U from the American Heart Association, Mid-Atlantic Affiliate and grants HL18292 and HL079087 from the NHLBI.


  1. 1.
    Duling BR, Berne RM (1970) Longitudinal gradients in periarteriolar oxygen tension. A possible mechanism for the participation of oxygen in local regulation of blood flow. Circ Res 27:669–678.PubMedGoogle Scholar
  2. 2.
    Pittman RN (2005) Oxygen transport and exchange in the microcirculation. Microcirculation 12:59–70.PubMedCrossRefGoogle Scholar
  3. 3.
    Tsai AG, Friesenecker B, Mazzoni MC, Kerger H, Buerk DG, Johnson PC, Intaglietta M (1998) Microvascular and tissue oxygen gradients in the rat mesentery. Proc Natl Acad Sci USA 95:6590–6595.PubMedCrossRefGoogle Scholar
  4. 4.
    Vanderkooi JM, Maniara G, Green TJ, Wilson DF (1987) An optical method for measurement of dioxygen concentration based upon quenching of phosphorescence. J Biol Chem 262:5476–5482.PubMedGoogle Scholar
  5. 5.
    Golub AS, Pittman RN (2008) PO2 measurements in the microcirculation using phosphorescence quenching microscopy at high magnification. Am J Physiol Heart Circ Physiol 294:H2905–H2916.PubMedCrossRefGoogle Scholar
  6. 6.
    Vadapalli AR, Pittman RN, Popel AS (2000) Estimating oxygen transport resistance of the microvascular wall. Am J Physiol Heart Circ Physiol 279:657–671.Google Scholar
  7. 7.
    Golub AS, Pittman RN (2005) Erythrocyte-associated transients in PO2 revealed in capillaries of rat mesentery. Am J Physiol Heart Circ Physiol 288:H2735–H2743.PubMedCrossRefGoogle Scholar
  8. 8.
    Hellums JD (1977) The resistance to oxygen transport in the capillaries relative to that in the surrounding tissue. Microvasc Res 13:131–136.PubMedCrossRefGoogle Scholar
  9. 9.
    Golub AS, Barker MC, Pittman RN (2007) PO2 profiles near arterioles and tissue oxygen consumption in the rat mesentery. Am J Physiol Heart Circ Physiol 293:H1097–H1106.PubMedCrossRefGoogle Scholar
  10. 10.
    Golub AS, Barker MC, Pittman RN (2008) Microvascular oxygen tension in the rat mesentery. Am J Physiol Heart Circ Physiol 294:H21–H28.PubMedCrossRefGoogle Scholar
  11. 11.
    Carvalho H, Pittman RN (2008) Longitudinal and radial gradients of PO2 in the hamster cheek pouch microcirculation. Microcirculation 15:215–224.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Roland N. Pittman
    • 1
  • Aleksander S. Golub
    • 1
  • Helena Carvalho
    • 1
  1. 1.Department of Physiology and BiophysicsVirginia Commonwealth UniversityRichmondUSA

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