The purpose of this study was to measure the hemoglobin oxygenation in retinal vessels and to evaluate the sensitivity and reproducibility of the measurement. Using a fundus camera equipped with a special dual wavelength transmission filter and a color charge-coupled device camera, two monochromatic fundus images at 548 and 610 nm were recorded simultaneously. The optical densities of retinal vessels for both wavelengths and their ratio, which is known to be proportional to the oxygen saturation, were calculated. From 50-deg images, the used semiautomatic vessel recognition and tracking algorithm recognized and measured vessels of 100 microm or more in diameter. On average, arterial and venous oxygen saturations were measured at 98+/-10.1% and 65+/-11.7%, respectively. For measurements in the same vessel segments from the five images per subject, standard deviations of 2.52% and 3.25% oxygen saturation were found in arteries and veins, respectively. Respiration of 100% oxygen increased the mean arterial and venous oxygen saturation by 2% and 7% respectively. A simple system for noninvasive optical oximetry, consisting of a special filter in a fundus camera and software, was introduced. It is able to measure the oxygen saturation in retinal branch vessels with reproducibility and sensitivity suitable for clinical investigations.

The increase of retinal vessel oxygen saturation in diabetic retinopathy points to a diabetic microvascular alteration. This may be due to occlusions and obliterations in the capillary bead and the formation of arterio-venous shunt vessels. On the other hand, hyperglycaemia-induced endothelial dysfunction, with subsequent suppression of the endothelial NO-synthase and disturbance of the vascular auto-regulation, may contribute to retinal tissue hypoxia.

Acta Ophthalmol. 2010: 88: 717–722 Abstract. The present article describes a standard instrument for the continuous online determination of retinal vessel diameters, the commercially available retinal vessel analyzer. This report is intended to provide informed guidelines for measuring ocular blood flow with this system. The report describes the principles underlying the method and the instruments currently available, and discusses clinical protocol and the specific parameters measured by the system. Unresolved questions and the possible limitations of the technique are also discussed.

OBJECTIVE—Stimulation of the retina with flickering light increases retinal vessel diameters in humans. Nitric oxide is a mediator of the retinal vasodilation to flicker. The reduction of vasodilation is considered an endothelial dysfunction. We investigated the response of retinal vessels to flickering light in diabetic patients in different stages of diabetic retinopathy. RESEARCH DESIGN AND METHODS—We studied 53 healthy volunteers, 68 type 1 diabetic patients, and 172 type 2 diabetic patients. The diameter of retinal vessels was measured continuously online with the Dynamic Vessel Analyzer (DVA). Diabetic retinopathy was classified using Early Treatment Diabetic Retinopathy Study criteria. Changes in vasodilation are expressed as percent change over baseline values. RESULTS—After adjustments for age, sex, and antihypertensive treatment, the response of retinal arterioles to diffuse luminance flicker was significantly diminished in patients with type 1 diabetes compared with healthy volunteers. The vasodilation of retinal arterioles and venules decreased continuously with increasing stages of diabetic retinopathy. The retinal arterial diameter change was 3.6 ± 2.1% in the control group, 2.6 ± 2.5% in the no diabetic retinopathy group, 2.0 ± 2.7% in the mild nonproliferative diabetic retinopathy (NPDR) group, 1.6 ± 2.2% in the moderate NPDR group, 1.8 ± 1.9% in severe NPDR group, and 0.8 ± 1.6% in proliferative diabetic retinopathy group. CONCLUSIONS—Flicker responses of retinal vessels are abnormally reduced in diabetic patients. This decreased response deteriorated with increasing stages of retinopathy. The response was already reduced before clinical appearance of retinopathy. The noninvasive testing of retinal autoregulation with DVA might prove to be of value in early detection of diabetic vessel pathological changes.

A significant correlation of age and bFR was not found in the small sample examined. Untreated arterial hypertension appeared to be associated with a reduced flicker response. The value of such functional vessel properties in the screening of vasosclerosis and in diagnostics in arterial hypertension should be examined in further studies.

OBJECTIVEFlicker light–induced retinal vasodilation may reflect endothelial function in the retinal circulation. We investigated flicker light–induced vasodilation in individuals with diabetes and diabetic retinopathy.RESEARCH DESIGN AND METHODSParticipants consisted of 224 individuals with diabetes and 103 nondiabetic control subjects. Flicker light–induced retinal vasodilation (percentage increase over baseline diameter) was measured using the Dynamic Vessel Analyzer. Diabetic retinopathy was graded from retinal photographs.RESULTSMean ± SD age was 56.5 ± 11.8 years for those with diabetes and 48.0 ± 16.3 years for control subjects. Mean arteriolar and venular dilation after flicker light stimulation were reduced in participants with diabetes compared with those in control subjects (1.43 ± 2.10 vs. 3.46 ± 2.36%, P < 0.001 for arteriolar and 2.83 ± 2.10 vs. 3.98 ± 1.84%, P < 0.001 for venular dilation). After adjustment for age, sex, diabetes duration, fasting glucose, cholesterol and triglyceride levels, current smoking status, systolic blood pressure, and use of antihypertensive and lipid-lowering medications, participants with reduced flicker light–induced vasodilation were more likely to have diabetes (odds ratio 19.7 [95% CI 6.5–59.1], P < 0.001 and 8.14 [3.1–21.4], P < 0.001, comparing lowest vs. highest tertile of arteriolar and venular dilation, respectively). Diabetic participants with reduced flicker light–induced vasodilation were more likely to have diabetic retinopathy (2.2 [1.2–4.0], P = 0.01 for arteriolar dilation and 2.5 [1.3–4.5], P = 0.004 for venular dilation).CONCLUSIONSReduced retinal vasodilation after flicker light stimulation is independently associated with diabetes status and, in individuals with diabetes, with diabetic retinopathy. Our findings may therefore support endothelial dysfunction as a pathophysiological mechanism underlying diabetes and its microvascular manifestations.

Abstract:We present a system capable of measuring the total retinal blood flow using a combination of dual beam Fourier-domain Doppler optical coherence tomography with orthogonal detection planes and a fundus camera-based retinal vessel analyzer. Our results show a high degree of conformity of venous and arterial flows, which corroborates the validity of the measurements. In accordance with Murray's law, the log-log regression coefficient between vessel diameter and blood flow was found to be ~3. The blood's velocity scaled linearly with the vessel diameter at higher diameters (> 60 µm), but showed a clear divergence from the linear dependence at lower diameters. Good agreement with literature data and the large range and high measurement sensitivity point to a high potential for further investigations. 673-800 (1987). 33. J. E. Grunwald, C. E. Riva, J. Baine, and A. J. Brucker, "Total retinal volumetric blood flow rate in diabetic patients with poor glycemic control," Invest. Ophthalmol. Vis. Sci. 33(2), 356-363 (1992). 34. J. E. Grunwald, J. DuPont, and C. E. Riva, "Retinal haemodynamics in patients with early diabetes mellitus," Br.J. Ophthalmol. 80(4), 327-331 (1996). 35. B. Pemp, E. Polska, G. Garhofer, M. Bayerle-Eder, A. Kautzky-Willer, and L. Schmetterer, "Retinal blood flow in type 1 diabetic patients with no or mild diabetic retinopathy during euglycemic clamp," Diabetes Care 33(9), 2038-2042 (2010 562-568 (1931). 39. T. W. Secomb and A. R. Pries, "Blood viscosity in microvessels: Experiment and theory," C. R. Phys. 14(6), 470-478 (2013). 40. E. Logean, L. Schmetterer, and C. E. Riva, "Velocity Profile of Red Blood Cells in Human Retinal Vessels using Confocal Scanning Laser Doppler Velocimetry," Laser Phys. 13, 45-51 (2003). 41. J. P. Garcia, Jr., P. T. Garcia, and R. B. Rosen, "Retinal blood flow in the normal human eye using the canon laser blood flowmeter," Ophthalmic Res. 34(5), 295-299 (2002)

The Retinal Vessel Analyzer (RVA) is a measuring device for online measurement of the diameter of retinal vessels in relation to time and locations along the vessel. It is furthermore provided with several tools for analyzing the measured data. The fundamental components consist of a fundus camera with CCD measuring camera attached and an advanced image-processing unit. The measurement range is from 90 microns, temporal resolution is 40 ms and measurement resolution is less than 1 micron. Systematic error of non-linearity is S < or = 1.6%, reproducibility is given by variation coefficient: short term vcs = 1.5%, long term vcl = 2.8%.