Out-of-focus signal rejection for pO measurements using two-photon phosphorescence lifetime microscopy.

Two-photon phosphorescence lifetime microscopy has been a key tool for studying cerebral oxygenation in mice. However, the accuracy of the partial pressure of oxygen (pO) measurements is affected by out-of-focus signal. In this work, we applied reconfigurable differential aberration imaging to characterize and correct for out-of-focus signal contamination in intravascular pO imaging. Our results show that signal contamination is higher in more oxygenated vessels and that it could be effectively removed using the proposed method.