Brain Tissue PO2 Measurement During Normoxia and Hypoxia Using Two-Photon Phosphorescence Lifetime Microscopy

  • Kui Xu
  • David A. Boas
  • Sava Sakadžić
  • Joseph C. LaMannaEmail author
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 977)


Key to the understanding of the principles of physiological and structural acclimatization to changes in the balance between energy supply (represented by substrate and oxygen delivery, and mitochondrial oxidative phosphorylation) and energy demand (initiated by neuronal activity) is to determine the controlling variables, how they are sensed and the mechanisms initiated to maintain the balance. The mammalian brain depends completely on continuous delivery of oxygen to maintain its function. We hypothesized that tissue oxygen is the primary sensed variable. In this study two-photon phosphorescence lifetime microscopy (2PLM) was used to determine and define the tissue oxygen tension field within the cerebral cortex of mice to a cortical depth of between 200–250 μm under normoxia and acute hypoxia (FiO2 = 0.10). High-resolution images can provide quantitative distributions of oxygen and intercapillary oxygen gradients. The data are best appreciated by quantifying the distribution histogram that can then be used for analysis. For example, in the brain cortex of a mouse, at a depth of 200 μm, tissue oxygen tension was mapped and the distribution histogram was compared under normoxic and mild hypoxic conditions. This powerful method can provide for the first time a description of the delivery and availability of brain oxygen in vivo.


Oxygen partial pressure 2PLM Tissue oxygen tension Distribution histogram Mouse 



This study was supported by the NIH grants R01 NS38632, R24 NS092986, R01 NS091230, R01 NS055104, and R01 EB021018.


  1. 1.
    LaManna JC (2007) Hypoxia in the central nervous system. Essays Biochem 43:139–151CrossRefPubMedGoogle Scholar
  2. 2.
    LaManna JC, Vendel LM, Farrell RM (1992) Brain adaptation to chronic hypobaric hypoxia in rats. J Appl Physiol 72:2238–2243PubMedGoogle Scholar
  3. 3.
    LaManna JC, Cordisco BR, Knuese DE et al (1994) Increased capillary segment length in cerebral cortical microvessels of rats exposed to 3 weeks of hypobaric hypoxia. Adv Exp Med Biol 345:627–632CrossRefPubMedGoogle Scholar
  4. 4.
    Ndubuizu O, LaManna JC (2007) Brain tissue oxygen concentration measurements. Antioxid Redox Signal 9(8):1207–1219CrossRefPubMedGoogle Scholar
  5. 5.
    Sakadžić S, Roussakis E, Yaseen MA et al (2010) Two-photon high-resolution measurement of partial pressure of oxygen in cerebral vasculature and tissue. Nat Methods 7(9):755–759CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Finikova OS, Lebedev AY, Aprelev A et al (2008) Oxygen microscopy by two-photon-excited phosphorescence. ChemPhysChem 9(12):1673–1679CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Lecoq J, Parpaleix A, Roussakis E et al (2011) Simultaneous two-photon imaging of oxygen and blood flow in deep cerebral vessels. Nat Med 17(7):893–898CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Sakadžić S, Mandeville ET, Gagnon L et al (2014) Large arteriolar component of oxygen delivery implies a safe margin of oxygen supply to cerebral tissue. Nat Commun 5:5734CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Baraghis E, Devor A, Fang Q et al (2011) Two-photon microscopy of cortical NADH fluorescence intensity changes: correcting contamination from the hemodynamic response. J Biomed Opt 16(10):106003CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Lauro KL, LaManna JC (1997) Adequacy of cerebral vascular remodeling following three weeks of hypobaric hypoxia. Examined by an integrated composite analytical model. Adv Exp Med Biol 411:369–376CrossRefPubMedGoogle Scholar
  11. 11.
    Dunn JF, Grinberg O, Roche M et al (2000) Noninvasive assessment of cerebral oxygenation during acclimation to hypobaric hypoxia. J Cereb Blood Flow Metab 20(12):1632–1635CrossRefPubMedGoogle Scholar

Copyright information

© Springer International Publishing AG 2017

Authors and Affiliations

  • Kui Xu
    • 1
  • David A. Boas
    • 2
  • Sava Sakadžić
    • 2
  • Joseph C. LaManna
    • 1
    Email author
  1. 1.Department of Physiology and BiophysicsCase Western Reserve University, School of MedicineClevelandUSA
  2. 2.Optics Division, Athinoula A. Martinos Center for Biomedical Imaging, Department of RadiologyMassachusetts General Hospital and Harvard Medical SchoolCharlestownUSA

Personalised recommendations