Two-photon high-resolution measurement of partial pressure of oxygen in cerebral vasculature and tissue

Sava Sakadžić, Emmanuel Roussakis, Mohammad A. Yaseen, Emiri T. Mandeville, Vivek Srinivasan, Ken Arai, Svetlana Ruvinskaya, Anna Devor, Eng H. Lo, Sergei A. Vinogradov, David A. Boas

Research output: Contribution to journalArticlepeer-review

342 Scopus citations


Measurements of oxygen partial pressure (pO2) with high temporal and spatial resolution in three dimensions is crucial for understanding oxygen delivery and consumption in normal and diseased brain. Among existing pO 2 measurement methods, phosphorescence quenching is optimally suited for the task. However, previous attempts to couple phosphorescence with two-photon laser scanning microscopy have faced substantial difficulties because of extremely low two-photon absorption cross-sections of conventional phosphorescent probes. Here we report to our knowledge the first practical in vivo two-photon high-resolution pO2 measurements in small rodents' cortical microvasculature and tissue, made possible by combining an optimized imaging system with a two-photon-enhanced phosphorescent nanoprobe. The method features a measurement depth of up to 250 μm, sub-second temporal resolution and requires low probe concentration. The properties of the probe allowed for direct high-resolution measurement of cortical extravascular (tissue) pO 2, opening many possibilities for functional metabolic brain studies.

Original languageEnglish (US)
Pages (from-to)755-759
Number of pages5
JournalNature Methods
Issue number9
StatePublished - Sep 1 2010
Externally publishedYes

ASJC Scopus subject areas

  • Biotechnology
  • Molecular Biology
  • Biochemistry
  • Cell Biology
  • Medicine(all)


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