Accessibility to the bulbar conjunctival microvasculature provides a means to assess blood supply to the cerebral cortex and thus optimize therapeutic interventions designed to prevent or reduce the risk of cerebral vascular disease and stroke. The feasibility of a method for quantitative measurements of conjunctiva blood vessel diameter, blood velocity, and flow in the human eye is reported. The method is based on slit lamp biomicroscope digital imaging coupled with a space time image analysis technique. A sequence of conjunctiva microvasculature images were captured at a rate of 50 Hz. The images were analyzed to determine blood vessel diameter, velocity and flow. Blood vessel diameter measurements ranged between 8.7 and 24.3 microns, with a mean value of 15.5 microns. Blood flow rate ranged between 27.3 and 296.9 pl/s, with a mean value of 111.8 pl/s. The relationship between blood flow and vessel diameter was fit with a power law curve (R = 0.87). The application of this technique for in vivo quantitative assessment of blood flow dynamics has potential to impact diagnosis and monitoring of various cardiovascular and blood disorders.

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Oxygen extraction fraction before and during light flicker stimulation is reported in human subjects for the first time. Oxygen extraction fraction decreased during light flicker stimulation, indicating the change in DO2 exceeded that of MO2. This technology is potentially useful for the detection of changes in OEF response to light flicker in physiological and pathological retinal conditions.

DO2_IR and MO2_IR were maintained during moderate hypoxia, but reduced under severe hypoxia, indicating blood flow compensation became insufficient for the reduced oxygen availability. Future studies may aid our understanding of retinal metabolic function in ischemic conditions.

Purpose An optical section phosphorescence lifetime imaging system was developed for three-dimensional mapping of oxygen tension (PO2) in chorioretinal vasculatures. Methods A laser line was projected at an oblique angle and scanned on the retina after intravenous injection of an oxygen-sensitive molecular probe to generate phosphorescence optical section images. An automated software algorithm segmented and combined images from spatially adjacent locations to construct depth-displaced en face retinal images. Intravascular PO2 was measured by determining the phosphorescence lifetime. Three-dimensional chorioretinal PO2 maps were generated in rat eyes under varying fractions of inspired oxygen. Results Under an air-breathing condition, mean PO2 in the choroid, retinal arteries, capillaries, and veins were 58 ± 2 mm Hg, 47 ± 2 mm Hg, 44 ± 2 mm Hg, and 35 ± 2 mm Hg, respectively. The mean arteriovenous PO2 difference was 12 ± 2 mm Hg. With a lower fraction of inspired oxygen, chorioretinal vascular PO2 and mean arteriovenous PO2 differences decreased compared with measurements under an air-breathing condition. Retinal venous PO2 was statistically lower than PO2 measured in the retinal artery, capillaries, and choroid (P < 0.004). Conclusions Three-dimensional mapping of chorioretinal oxygen tension allowed quantitative PO2 measurements in large retinal blood vessels and in retinal capillaries. This method has the potential to facilitate better understanding of retinal oxygenation in health and disease.

PurposeTo test the hypothesis that retinal vascular diameter and hemoglobin oxygen saturation alterations, according to stages of diabetic retinopathy (DR), are discernible with a commercially available scanning laser ophthalmoscope (SLO).MethodsOne hundred eighty-one subjects with no diabetes (No DM), diabetes with no DR (No DR), nonproliferative DR (NPDR), or proliferative DR (PDR, all had photocoagulation) underwent imaging with an SLO with dual lasers (532 nm and 633 nm). Customized image analysis software determined the diameters of retinal arteries and veins (DA and DV) and central retinal artery and vein equivalents (CRAE and CRVE). Oxygen saturations of hemoglobin in arteries and veins (SO2A and SO2V) were estimated from optical densities of vessels on images at the two wavelengths. Statistical models were generated by adjusting for effects of sex, race, age, eye, and fundus pigmentation.ResultsDA, CRAE, and CRVE were reduced in PDR compared to No DM (P ≤ 0.03). DV and CRVE were similar between No DM and No DR, but they were higher in NPDR than No DR (P ≤ 0.01). Effect of stage of disease on SO2A differed by race, being increased relative to No DM in NPDR and PDR in Hispanic participants only (P ≤ 0.02). Relative to No DM, SO2V was increased in NPDR and PDR (P ≤ 0.05).ConclusionsAlterations in retinal vascular diameters and SO2 by diabetic retinopathy stage can be detected with a widely available SLO, and covariates such as race can influence the results.

The known biophysical variations of hemoglobin (Hb) S and Hb C may result in hemodynamic differences between subjects with SS and SC disease. The purpose of this study was to measure and compare conjunctival hemodynamics between subjects with Hb SS and SC hemoglobinopathies. Image sequences of the conjunctival microcirculation were acquired in 9 healthy control subjects (Hb AA), 24 subjects with SC disease, and 18 subjects with SS disease, using a prototype imaging system. Diameter (D) and blood velocity (V) measurements were obtained in multiple venules of each subject. Data were categorized according to venule caliber by averaging V and D for venules with diameters less than (vessel size 1) or greater than (vessel size 2) 15 µm. V in vessel size 2 was significantly greater than V in vessel size 1 in the AA and SS groups (P ≥ 0.009), but not in the SC group (P = 0.1). V was significantly lower in the SC group as compared to the SS group (P = 0.03). In AA and SS groups, V correlated with D (P ≥ 0.005), but the correlation was not statistically significant in the SC group (P = 0.08). V was inversely correlated with hematocrit in the SS group for large vessels (P = 0.03); however, no significant correlation was found in the SC group (P ≥ 0.2). Quantitative assessment of conjunctival microvascular hemodynamics in SS and SC disease may advance understanding of sickle cell disease pathophysiology and thereby improve therapeutic interventions.

The conjunctival microcirculation is accessible for direct visualization and quantitative assessment of microvascular hemodynamic properties. Currently available methods to assess hemodynamics in the conjunctival microvasculature use manual or semi-automated algorithms, which can be inefficient for application to a large number of microvessels within the microvascular network. We present an automated image analysis method for measurements of diameter and blood velocity in microvessels. The method was applied to conjunctival microcirculation images acquired in 15 healthy human subjects. Frangi filtering, thresholding, and morphological closing were applied to automatically segment microvessels, while variance filtering was used to detect blood flow. Diameter and blood velocity were measured in arterioles and venules within the conjunctival microvascular network, and blood flow and wall shear rate were calculated. Repeatability and validity of hemodynamic measurements were established. The automated image analysis method allows reliable, rapid and quantitative assessment of hemodynamics in the conjunctival microvascular network and can be potentially applied to microcirculation images of other tissues.