Two-photon phosphorescence lifetime microscopy of retinal capillary plexus oxygenation in mice

Ikbal Şencan, Tatiana V. Esipova, Mohammad A. Yaseen, Buyin Fu, David A. Boas, Sergei A. Vinogradov, Mahnaz Shahidi, Sava Sakadžić

Research output: Contribution to journalArticlepeer-review

15 Scopus citations


Impaired oxygen delivery and/or consumption in the retinal tissue underlies the pathophysiology of many retinal diseases. However, the essential tools for measuring oxygen concentration in retinal capillaries and studying oxygen transport to retinal tissue are still lacking. We show that two-photon phosphorescence lifetime microscopy can be used to map absolute partial pressures of oxygen (pO 2) in the retinal capillary plexus. Measurements were performed at various retinal depths in anesthetized mice under systemic normoxic and hyperoxic conditions. We used a newly developed two-photon phosphorescent oxygen probe, based on a two-photon absorbing platinum tetraphthalimidoporphyrin, and commercially available optics without correction for optical aberrations of the eye. The transverse and axial distances within the tissue volume were calibrated using a model of the eye's optical system. We believe this is the first demonstration of in vivo depth-resolved imaging of pO 2 in retinal capillaries. Application of this method has the potential to advance our understanding of oxygen delivery on the microvascular scale and help elucidate mechanisms underlying various retinal diseases.

Original languageEnglish
Article number126501
JournalJournal of biomedical optics
Issue number12
StatePublished - Dec 1 2018


  • oxygen partial pressure
  • phosphorescence quenching
  • retinal imaging
  • two-photon microscopy


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