J. Ning

Oxygen Saturation in Optic Nerve Head Structures by Hyperspectral Image Analysis

Purpose: A method is presented for the calculation and visualization of percent blood oxygen saturation from specific tissue structures in hyperspectral images of the optic nerve head (ONH). Methods: Trans-pupillary images of the primate optic nerve head and overlying retinal blood vessels were obtained with a hyperspectral imaging (HSI) system attached to a fundus camera. Images were recorded during normal blood flow and after partially interrupting flow to the ONH and retinal circulation by elevation of the intraocular pressure (IOP) from 10 mmHg to 55 mmHg in steps. Percent oxygen saturation was calculated from groups of pixels associated with separate tissue structures, using a linear least-squares curve fit of the recorded hemoglobin spectrum to reference spectra obtained from fully oxygenated and deoxygenated red cell suspensions. Color maps of saturation were obtained from a new algorithm that enables comparison of oxygen saturation from large vessels and tissue areas in hyperspectral images. Results: Percent saturation in retinal vessels and from the average over ONH structures (IOP = 10 mmHg) was (mean ± SE): artery 81.8 ± 0.4%, vein 42.6 ± 0.9%, average ONH 68.3 ± 0.4%. Raising IOP from 10 mmHg to 55 mmHg for 5 min caused blood oxygen saturation to decrease (mean ± SE): artery 46.1 ± 6.2%, vein 36.1 ± 1.6%, average ONH 41.9 ± 1.6%. The temporal cup showed the highest saturation at low and high IOP (77.3 ± 1.0% and 60.1 ± 4.0%) and the least reduction in saturation at high IOP (22.3%) compared with that of the average ONH (38.6%). A linear relationship was found between saturation indices obtained from the algorithm and percent saturation values obtained by spectral curve fits to calibrated red cell samples. Conclusions: Percent oxygen saturation was determined from hyperspectral images of the ONH tissue and retinal vessels overlying the ONH at normal and elevated IOP. Pressure elevation was shown to reduce blood oxygen saturation in vessels and ONH structures, with the smallest reduction in the ONH observed in the temporal cup. IOP-induced saturation changes were visualized in color maps using an algorithm that follows saturation-dependent changes in the blood spectrum and blood volume differences across tissue. Reduced arterial saturation at high IOP may have resulted from a flow-dependent mechanism.

Algorithms for automated oximetry along the retinal vascular tree from dual-wavelength fundus images.

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.

Hyperspectral Algorithm for Mapping Tissue Oxygen Saturation

A simple algorithm based on computational estimates of spectral feature amplitudes in hyperspectral images is described for mapping oxygen delivery in tissue. Features are determined from contiguous image bands spanning the prominent absorption wavelengths of hemoglobin. Pixel values along the z-axis at each image coordinate establish a spectral curve. A reference baseline is obtained by linear interpolation between isosbestic points on the curve which are insensitive to changes in oxygen saturation of the blood. An oxygen-sensitive vector is determined from areas between the recorded curve and baseline, which estimates relative amounts of oxyhemoglobin and deoxyhemoglobin that contribute to the blood spectrum. Corrections are applied to account for variation in total light between recordings, and for the volume of blood at each pixel location in the recording. Here we apply the method to map relative oxygen saturation in the optic nerve head and overlying retinal vessels at normal and elevated intraocular pressure

Retinal oxygen saturation evaluation by multi-spectral fundus imaging

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.

Oxygen saturation changes in the optic nerve head during acute intraocular pressure elevation in monkeys

Background and Objective: To evaluate the effect of an acute elevated intraocular pressure (IOP) on oxygen saturation of structures of the optic nerve head. Study Design/Materials and Methods: In the cynomolgus monkey eye, IOP was set to 10 mm Hg, and then raised to 30, 45, and 55 mm Hg. The ONH and overlying vessels were imaged using a fundus camera attached to a hyperspectral imaging system (HSI) at 10 and 30 minutes after IOP elevation. Results: Raising IOP from 10 to 30 mm Hg did not significantly (P < 0.0001) change saturation in vessels or ONH tissue structures but at 55 mm Hg, all structures showed significant reduction. Conclusions: Quantitative assay of the blood oxygen saturation in structures on the surface and overlying the optic nerve head is possible using hyperspectral imaging techniques.