Purpose: To develop a multi-spectral method to measure oxygen saturation of the retina in the human eye.
Methods: Five Cynomolgus monkeys with normal eyes were anesthetized with intramuscular ketamine/xylazine and intravenous pentobarbital. Multi-spectral fundus imaging was performed in five monkeys with a commercial fundus camera equipped with a liquid crystal tuned filter in the illumination light path and a 16-bit digital camera. Recording parameters were controlled with software written specifically for the application. Seven images at successively longer oxygen-sensing wavelengths were recorded within 4 seconds. Individual images for each wavelength were captured in less than 100 msec of flash illumination. Slightly misaligned images of separate wavelengths due to slight eye motion were registered and corrected by translational and rotational image registration prior to analysis. Numerical values of relative oxygen saturation of retinal arteries and veins and the underlying tissue in between the artery/vein pairs were evaluated by an algorithm previously described, but which is now corrected for blood volume from averaged pixels (n > 1000). Color saturation maps were constructed by applying the algorithm at each image pixel using a Matlab script.
Results: Both the numerical values of relative oxygen saturation and the saturation maps correspond to the physiological condition, that is, in a normal retina, the artery is more saturated than the tissue and the tissue is more saturated than the vein. With the multi-spectral fundus camera and proper registration of the multi-wavelength images, we were able to determine oxygen saturation in the primate retinal structures on a tolerable time scale which is applicable to human subjects.
Conclusions: Seven wavelength multi-spectral imagery can be used to measure oxygen saturation in retinal artery, vein, and tissue (microcirculation). This technique is safe and can be used to monitor oxygen uptake in humans.
This work is original and is not under consideration for publication elsewhere.
We present an automated method to perform accurate, rapid, and objective measurement of the blood oxygen saturation over each segment of the retinal vascular hierarchy from dual-wavelength fundus images. Its speed and automation (2 s per entire image versus 20 s per segment for manual methods) enables detailed level-by-level measurements over wider areas. An automated tracing algorithm is used to estimate vessel centerlines, thickness, directions, and locations of landmarks such as bifurcations and crossover points. The hierarchical structure of the vascular network is recovered from the trace fragments and landmarks by a novel algorithm. Optical densities (OD) are measured from vascular segments using the minimum reflected intensities inside and outside the vessel. The OD ratio (ODR=OD600/OD570) bears an inverse relationship to systemic HbO2 saturation (SO2). The sensitivity for detecting saturation change when breathing air versus pure oxygen was calculated from the measurements made on six subjects and was found to be 0.0226 ODR units, which is in good agreement with previous manual measurements by the dual-wavelength technique, indicating the validity of the automation. A fully automated system for retinal vessel oximetry would prove useful to achieve early assessments of risk for progression of disease conditions associated with oxygen utilization.