Exploratory study of non-invasive, high-resolution functional macular imaging in subjects with diabetic retinopathy.

AIM: To evaluate a high-resolution functional imaging device that yields quantitative data regarding macular blood flow and capillary network features in eyes with diabetic retinopathy (DR). METHODS: Prospective, cross-sectional comparative case-series in which blood flow velocities (BFVs) and non-invasive capillary perfusion maps (nCPMs) in macular vessels were measured in patients with DR and in healthy controls using the Retinal Functional Imager (RFI) device. RESULTS: A total of 27 eyes of 21 subjects were studied [9 eyes nonproliferative diabetic retinopathy (NPDR), 9 eyes proliferative diabetic retinopathy (PDR) and 9 controls]. All diabetic patients were type 2. All patients with NPDR and 5 eyes with PDR also had diabetic macular edema (DME). The NPDR group included eyes with severe (n=3) and moderate NPDR (n=6), and were symptomatic. A significant decrease in venular BFVs was observed in the macular region of PDR eyes when compared to controls (2.61±0.6 mm/s and 2.92±0.72 mm/s in PDR and controls, respectively, P=0.019) as well as PDR eyes with DME compared to NPDR eyes (2.36±0.51 mm/s and 2.94±1.09 mm/s in PDR with DME and NPDR, respectively, P=0.01). CONCLUSION: The RFI, a non-invasive imaging tool, provides high-resolution functional imaging of the retinal microvasculature and quantitative measurement of BFVs in visually impaired DR patients. The isolated diminish venular BFVs in PDR eyes compared to healthy eyes and PDR eyes with DME in comparison to NPDR eyes may indicate the possibility of more retinal vein compromise than suspected in advanced DR.

Non-invasive measurement techniques for quantitative assessment of optic nerve head blood flow.

Diseases of the optic nerve head involving changes in blood flow are common. However, the pathophysiology is not always fully understood. Several non-invasive methods for measuring optic nerve head blood flow are available, but currently no gold standard has been established. Methods for measuring blood flow in optic neuropathies including colour Doppler imaging, retinal function imager, optical coherence tomography angiography and laser speckle flowgraphy are reviewed. Ultrasound colour Doppler imaging is a fast measurement technique where several different parameters, especially the blood flow velocity, can be calculated. Though used for many years in ophthalmology, its use is not standardized and it requires significant observer skills. The retinal function imager is a direct method where the haemoglobin in erythrocytes is visualized and blood flow velocities in retinal vessels are calculated from a series of photos. The technique is not suitable for direct measurement of blood flow within the optic nerve head. Laser speckle flowgraphy uses a laser light which creates a light scatter pattern in the tissue. Particles moving in the area causes changes in the speckle pattern from which a relative blood flow can be estimated. It is, however, not known whether optic nerve head microcirculation is measurable with the technique. Optical coherence tomography angiography uses multiple scans to evaluate blood flow with good reproducibility but often problems with artefacts. The technique is continuously being refined and increasingly used in research as a tool for the study of blood flow in retinopathies and optic neuropathies. Most of the conducted studies are based on small sample sizes, but some of the methods show promising results in an optic nerve head blood flow research setting. Further and larger studies are required to provide standardized and comparable measurements before one or more of the methods can be considered clinical helpful in daily practice.

Microcirculation in the conjunctiva and retina in healthy subjects.

BACKGROUND: The aim was to determine the relationship between bulbar conjunctival microcirculation and retinal microcirculation in a healthy population. METHOD: A functional slit-lamp biomicroscope (FSLB) was used to measure blood flow velocity (BFV) and blood flow rate (BFR) in the conjunctiva while a retinal function imager (RFI) was used to measure macular BFV and BFR in the retina. One eye of each subject of 58 self-reported healthy subjects was imaged in the same session on the same day. RESULTS: The mean BFV in the venules of the conjunctiva was 0.49 ± 0.13 mm/s, which was significantly slower than that in the retinal arterioles (3.71 ± 0.78 mm/s, P < 0.001) and retinal venules (2.98 ± 0.58 mm/s, P < 0.001). The BFR in the conjunctiva (0.09 nl/s) was also significantly lower than that in the retina (arterioles = 0.81 nl/s, venules = 0.68 nl/s, all P < 0.001). The BFVs and BFRs were not related between the conjunctiva and retina (r ranged from - 0.17 to - 0.05, all P > 0.05). CONCLUSION: The microcirculation in the retina appeared to be different from that in the conjunctiva.

The effect of sildenafil on retinal blood velocity in healthy subjects.

PURPOSE: It has been suggested that Sildenafil may have beneficial therapeutic effects in the treatment of neurodegenerative disorders. The retinal circulation is of significant interest as a marker of cerebral vascular disease since the retinal and cerebral vasculatures share many morphological and physiological properties, yet only the retinal circulation can be directly visualized. Therefore, our aim was to assess the change induced by Sildenafil on retinal blood velocity. METHODS: Retinal flow velocity was measured 0.5, 3 and 6 h following administration of 100 mg of Sildenafil using the Retinal Function Imager. RESULTS: No clinical change in either systemic blood pressure or retinal flow velocities were observed. However, when controlling for heart rate and blood pressure, a significant drop in venous flow velocity 6 h following treatment (mean drop 0.3 ± 0.07; 95% CI: 0.44-0.56, P = 0.023) was revealed. CONCLUSIONS: In healthy volunteers, retinal venous flow velocity was significantly reduced at the 6-h time point following Sildenafil treatment. No effect was observed on arterial retinal flow velocity.

The inter-visit variability of retinal blood flow velocity measurements using retinal function imager (RFI).

BACKGROUND: To determine the inter-visit variability of retinal blood flow velocities (BFVs) using a retinal function imager (RFI) in healthy young subjects. METHODS: Twenty eyes of 20 healthy young subjects were enrolled. RFI imaging was performed to obtain the BFVs in retinal arterioles and venules in a field measuring 7.3 × 7.3 mm2 (setting: 35 degrees) centered on the fovea, and repeated measurements were obtained on two separate days. The inter-visit variability of BFVs was assessed by the concordance correlation coefficient (CCC) and coefficient of variance (CV). RESULTS: At the first visit, the mean BFV was 3.6 ± 0.8 mm/s and 3.0 ± 0.7 mm/s in arterioles and venules, respectively, which were not significantly different from those at the second visit (the BFV of arterioles was 3.5 ± 0.8 mm/s, and the BFV of venules was 3.0 ± 0.7 mm/s, P > 0.05, respectively). The CCC was 0.72 in the BFVs of arterioles and 0.67 in venules, and the CV was 10.8% in the BFVs of arterioles and 11.0% in venules. CONCLUSION: The inter-visit variability using the retinal function imager (RFI) with a large field of view appeared to be good and comparable to previously reported intra-visit and inter-eye variability.

Microvascular blood flow velocities measured with a retinal function imager: inter-eye correlations in healthy controls and an exploration in multiple sclerosis.

BACKGROUND: The retinal microcirculation has been studied in various diseases including multiple sclerosis (MS). However, inter-eye correlations and potential differences of the retinal blood flow velocity (BFV) remain largely unstudied but may be important in guiding eye selection as well as the design and interpretation of studies assessing or utilizing retinal BFV. The primary aim of this study was to determine inter-eye correlations in BFVs in healthy controls (HCs). Since prior studies raise the possibility of reduced BFV in MS eyes, a secondary aim was to compare retinal BFVs between MS eyes, grouped based on optic neuritis (ON) history and HC eyes. METHODS: Macular arteriole and venule BFVs were determined using a retinal function imager (RFI) in both eyes of 20 HCs. One eye from a total of 38 MS patients comprising 13 eyes with ON (MSON) and 25 eyes without ON (MSNON) history were similarly imaged with RFI. RESULTS: OD (right) and OS (left) BFVs were not significantly different in arterioles (OD: 3.95 ± 0.59 mm/s; OS: 4.08 ± 0.60 mm/s, P = 0.10) or venules (OD: 3.11 ± 0.46 mm/s; OS: 3.23 ± 0.52 mm/s, P = 0.06) in HCs. Very strong inter-eye correlations were also found between arteriolar (r = 0.84, P < 0.001) and venular (r = 0.87, P < 0.001) BFVs in HCs. Arteriolar (3.48 ± 0.88 mm/s) and venular (2.75 ± 0.53 mm/s) BFVs in MSNON eyes were significantly lower than in HC eyes (P = 0.009 and P = 0.005, respectively). Similarly, arteriolar (3.59 ± 0.69 mm/s) and venular (2.80 ± 0.45 mm/s) BFVs in MSON eyes were also significantly lower than in HC eyes (P = 0.046 and P = 0.048, respectively). Arteriolar and venular BFVs in MSON and MSNON eyes did not differ from each other (P = 0.42 and P = 0.48, respectively). CONCLUSIONS: Inter-eye arteriolar and venular BFVs do not differ significantly in HCs and are strongly correlated. Our findings support prior observations that arteriolar and venular BFVs may be reduced in MS eyes. Moreover, this seems to be the case in both MS eyes with and without a history of ON, raising the possibility of global blood flow alterations in MS. Future larger studies are needed to assess differences in BFVs between MSON and MSNON eyes.

Retinal nerve fiber layer (RNFL) integrity and its relations to retinal microvasculature and microcirculation in myopic eyes.

BACKGROUND: The aim was to determine retinal nerve fiber layer function and its relations to retinal microvasculature and microcirculation in patients with myopia. METHOD: Polarization-sensitive optical coherence tomography (PS-OCT) was used to measure phase retardation per unit depth (PR/UD, proportional to the birefringence) of the retinal nerve fiber layer (RNFL). Optical coherence tomography angiography (OCTA) was used to measure macular vessel density analyzed using fractal analysis. In addition, a retinal function imager (RFI) was used to measure macular blood flow velocities in arterioles and venules. Twenty-two patients with moderate myopia (MM, refraction > 3 and < 6 diopters), seventeen patients with high myopia (HM, ≥ 6 D) and 29 healthy control subjects (HC, ≤ 3.00 D) were recruited. One eye of each patient was imaged. RESULTS: Although the average PR/UD of the RNFL in the HM group did not reach a significant level, the birefringence of the inferior quadrant was significantly lower (P < 0.05) in the HM group compared to the HC group. Significant thinning of the average RNFL and focal thinning of RFNL in temporal, superior and inferior quadrants in the HM group were found, compared to the HC group (P < 0.05). There were no significant differences of retinal blood flow velocities in arterioles and venules among groups (P > 0.05). The macular vessel density in both superficial and deep vascular plexuses was significantly lower in the HM group than in the other two groups (P < 0.05) as well as in the MM group than in the HC group (P < 0.05). The average PR/UD and PR/UD in the inferior quadrant were not related to refraction, axial length, blood flow velocities and macular vessel densities (r ranged from - 0.09 to 0.19, P > 0.05). CONCLUSIONS: The impairment of the retinal nerve fiber birefringence in the HM group may be one of the independent features in high myopic eyes, which appeared not to relate to macular microvascular density and blood flow velocity.

The retinal function imager and clinical applications.

BACKGROUND: The Retinal Function Imager (RFI) provides in vivo and noninvasive imaging of both the retinal structure and function. REVIEW: The RFI can create capillary perfusion maps, measure blood flow velocity, and determine metabolic function including blood oximetry. It can aid clinical diagnosis as well as assess treatment response in several retinal vascular diseases including diabetic retinopathy. Blood flow velocity abnormalities have also been implicated in disease such as age-related macular degeneration and require further investigation. Compared with optical coherence tomography angiography, the RFI produces capillary maps of comparable image quality and wider field of view but it is unable to provide depth-resolved information and has longer image acquisition time. Currently, functional imaging using blood oximetry has limited applications and additional research is required. CONCLUSION: The RFI offers noninvasive, high-resolution imaging of retinal microvasculature by creating capillary perfusion maps. In addition, it is capable of measuring retinal blood velocity directly and performs functional imaging with retinal blood oximetry. Its clinical applications are broad and additional research with functional imaging may potentially lead to diagnosis of diseases and their progression before anatomic abnormalities become evident, but longer image acquisition times may limit its clinical adoption.

Retinal tissue hypoperfusion in patients with clinical Alzheimer's disease.

BACKGROUND: It remains unknow whether retinal tissue perfusion occurs in patients with Alzheimer's disease. The goal was to determine retinal tissue perfusion in patients with clinical Alzheimer's disease (CAD). METHODS: Twenty-four CAD patients and 19 cognitively normal (CN) age-matched controls were recruited. A retinal function imager (RFI, Optical Imaging Ltd., Rehovot, Israel) was used to measure the retinal blood flow supplying the macular area of a diameter of 2.5 mm centered on the fovea. Blood flow volumes of arterioles (entering the macular region) and venules (exiting the macular region) of the supplied area were calculated. Macular blood flow was calculated as the average of arteriolar and venular flow volumes. Custom ultra-high-resolution optical coherence tomography (UHR-OCT) was used to calculate macular tissue volume. Automated segmentation software (Orion, Voxeleron LLC, Pleasanton, CA) was used to segment six intra-retinal layers in the 2.5 mm (diameter) area centered on the fovea. The inner retina (containing vessel network), including retinal nerve fiber layer (RNFL), ganglion cell-inner plexiform layer (GCIPL), inner nuclear layer (INL) and outer plexiform layer (OPL), was segmented and tissue volume was calculated. Perfusion was calculated as the flow divided by the tissue volume. RESULTS: The tissue perfusion in CAD patients was 2.58 ± 0.79 nl/s/mm3 (mean ± standard deviation) and was significantly lower than in CN subjects (3.62 ± 0.44 nl/s/mm3, P <  0.01), reflecting a decrease of 29%. The flow volume was 2.82 ± 0.92 nl/s in CAD patients, which was 31% lower than in CN subjects (4.09 ± 0.46 nl/s, P <  0.01). GCIPL tissue volume was 0.47 ± 0.04 mm3 in CAD patients and 6% lower than CN subjects (0.50 ± 0.05 mm3, P < 0.05). No other significant alterations were found in the intra-retinal layers between CAD and CN participants. CONCLUSIONS: This study is the first to show decreased retinal tissue perfusion that may be indicative of diminished tissue metabolic activity in patients with clinical Alzheimer's disease.

Application of retinal functional imager in retinal diseases / 眼科新进展

Newer retinal imaging technologies help us in understanding the pathogenesis of many retinal pathologies,such as diabetic retinopathy,age related macular degeneration,glaucoma and uveitis.Early detection of these retinal diseases can prevent the onset of progressive vision loss,and aid in the development of new treatment options.Retinal functional imager (RFI) is an unique and noninvasive functional imaging system.Unlike most of the available newer retinal imaging tools,the RFI not only shows retinal structural changes,but can directly monitor functional changes and measure hemodynamic parameters,such as retinal bloodflow velocity,oximetric state,etc.This article reviews the utility progress of RFI in various retinal diseases.

Impaired retinal microcirculation in multiple sclerosis.

BACKGROUND: The transparent ocular structure enables quantitative analysis of microvasculature of retina, a neuronal tissue affected by multiple sclerosis (MS). OBJECTIVE: The aim of this study was to determine whether the retinal blood flow velocity and flow volume at the macula are impaired in patients with relapsing remitting multiple sclerosis (RRMS). METHODS: A total of 17 RRMS patients and 17 age- and gender-matched healthy subjects were assessed. A retinal function imager was used to measure the blood flow velocity of retinal arterioles and venules and to calculate the total perifoveal blood flow volume. RESULTS: The blood flow velocities of the retinal arterioles (3.34 ± 0.89 mm/s) and venules (2.61 ± 0.6 mm/s) were significantly lower in MS patients than normal subjects (arteriole: 4.10 ± 0.87 mm/s; venule: 3.22 ± 0.65 mm/s, both p = 0.01). In addition, the total perifoveal blood flow volume in arterioles (3.74 ± 1.64 nL/s) and venules (3.81 ± 1.60 nL/s) were significantly lower in MS patients than in normal subjects (arteriole: 4.87 ± 1.41 nL/s, p = 0.02; venule: 4.71 ± 1.64 nL/s, p = 0.04). CONCLUSION: The impaired retinal microcirculation in RRMS patients indicates microvascular dysfunction in MS.

Interactive retinal blood flow analysis of the macular region.

The study of retinal hemodynamics plays an important role to understand the onset and progression of diabetic retinopathy. In this work, we developed an interactive retinal analysis tool to quantitatively measure the blood flow velocity (BFV) and blood flow rate (BFR) in the macular region using the Retinal Function Imager (RFI). By employing a high definition stroboscopic fundus camera, the RFI device is able to assess retinal blood flow characteristics in vivo. However, the measurements of BFV using a user-guided vessel segmentation tool may induce significant inter-observer differences and BFR is not provided in the built-in software. In this work, we have developed an interactive tool to assess the retinal BFV and BFR in the macular region. Optical coherence tomography data was registered with the RFI image to locate the fovea accurately. The boundaries of the vessels were delineated on a motion contrast enhanced image and BFV was computed by maximizing the cross-correlation of pixel intensities in a ratio video. Furthermore, we were able to calculate the BFR in absolute values (µl/s). Experiments were conducted on 122 vessels from 5 healthy and 5 mild non-proliferative diabetic retinopathy (NPDR) subjects. The Pearson's correlation of the vessel diameter measurements between our method and manual labeling on 40 vessels was 0.984. The intraclass correlation (ICC) of BFV between our proposed method and built-in software was 0.924 and 0.830 for vessels from healthy and NPDR subjects, respectively. The coefficient of variation between repeated sessions was reduced significantly from 22.5% to 15.9% in our proposed method (p<0.001).

Assessment of potential vessel segmentation pitfalls in the analysis of blood flow velocity using the Retinal Function Imager.

PURPOSE: The purpose of our study was to investigate the potential pitfalls associated with different vessel segmentation methods using the built-in software of the Retinal Function Imager (RFI) for the analysis of retinal blood flow velocity (BFV). METHODS: Ten eyes of nine healthy subjects were enrolled in the study. Retinal blood flow measurements were obtained with the RFI device with a 20° field of view imaging. The same grader segmented the retinal vasculature using the RFI software in both sessions, with segments ranging in length from 50 to 100 pixels ("short segments") and 100-200 pixels ("long segments"). The blood flow velocities for the arteriolar and venular system were calculated, and the percentage of excluded vessel segments with high coefficients of variation (>45 %) was recorded and compared by paired t test. Spearman's correlation was used to analyze the relationship between measurements by the two vessel segmentation methods. RESULTS: The number of analyzed vessel segments did not differ significantly between the two groups (28.6 ± 2.6 short and 26.7 ± 4.6 long segments), while the percentage of acceptable segments was significantly higher in the long segment group (65.2 ± 11.4 % vs 85.2 ± 5.87 %, p = 0.001). In the short segment group, more than 15 % of vessel segments were rejected in all subjects, while in the long segment group only three subjects had a rejection rate of greater than 15 % (16.7 %, 18.7 % and 28 %). Both arteriolar and venular velocities were lower in the short segment group, although it reached significance only for arteriolar velocities (3.93 ± 0.55 vs. 4.45 ± 0.76 mm/s, p = 0.036 and 2.95 ± 0.56 vs. 3.17 ± 0.84 mm/s, p = 0.201 for arterioles and venules, respectively). Only venular velocities showed significant correlation (p = 0.003, R (2) = 0.67) between the two groups. CONCLUSIONS: Our results suggest that BFV measurements by the RFI may be affected by segment length, and care should therefore be taken in choosing vessel segment lengths used during the analysis of RFI data. Long segments of 100-200 pixels (400-800 µm) seem to provide more robust measurements, which can be explained by the analysis methodology of the RFI device.

Assesment of Conjunctival Microangiopathy in a Patient with Diabetes Mellitus Using the Retinal Function Imager.

Diabetes mellitus (DM) is notorious for causing retinal microangiopathy, but bulbar conjunctival microangiopathy (CM) mirroring the established retinal vessel changes, has also been observed. Recent studies suggest that CM occurs in all DM patients in various degrees depending on disease severity and occur even before non-proliferative retinopathy develops. Thus, CM might provide a means of early detection or even form a basis for early intervention of disease progression in DM patients. Herein we present - to our knowledge for the first time-the feasibility and applicability in diagnostic imaging of CM in a diabetic patient using a commercially available Retinal Function Imager (RFI, Optical Imaging Ltd, Rehovot, Israel).