A method for volumetric retinal tissue oxygen tension imaging

Inadequate retinal oxygenation occurs in many vision-threatening retinal diseases, including diabetic retinopathy, retinal vascular occlusions, and age-related macular degeneration. Therefore, techniques that assess retinal oxygenation are necessary to understand retinal physiology in health and disease. The purpose of the current study is to report a method for the three-dimensional (3D) imaging of retinal tissue oxygen tension (tPO₂) in rats.

Imaging was performed in Long Evans pigmented rats under systemic normoxia (N = 6) or hypoxia (N = 3). A vertical laser line was horizontally scanned on the retina and a series of optical section phase-delayed phosphorescence images were acquired. From these images, phosphorescence volumes at each phase delay were constructed and a 3D retinal tPO₂ volume was generated. Retinal tPO₂ volumes were quantitatively analyzed by generating retinal depth profiles of mean tPO₂ (M_tPO2) and the spatial variation of tPO₂ (SV_tPO2). The effects of systemic condition (normoxia/hypoxia) and retinal depth on M_tPO2 and SV_tPO2 were determined by mixed linear model.

Each 3D retinal tPO₂ volume was approximately 500 × 750 × 200 μm (horizontal × vertical × depth) and consisted of 45 en face tPO₂ images through the retinal depth. M_tPO2 at the chorioretinal interface was significantly correlated with systemic arterial oxygen tension (P = 0.007; N = 9). There were significant effects of both systemic condition and retinal depth on M_tPO2 and SV_tPO2, such that both were lower under hypoxia than normoxia and higher in the outer retina than inner retina (P < 0.001).

For the first time, 3D imaging of retinal tPO₂ was demonstrated, with potential future application for assessment of physiological alterations in animal models of retinal diseases.