We report the implementation and demonstration of an imaging system that combines two-photon phosphorescence lifetime microscopy (2PLM) and adaptive optics for improving oxygen partial pressure (pO2) measurement accuracy in deep cortical layers of mice. The technique retrieves the background signal by forming an aberrated torus focus on the sample plane with an optical phase mask imposed on the system wavefront and subtracts it from normal phosphorescence emission signal. The proposed method is validated with intravascular pO2 measurements from six mice imaged at up to ~600 μm depth.
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