Retinal vascular dynamics: A window for observing an irregular heartbeat. A case report.

OBJECTIVE: We aimed to characterize several aspects of retinal vascular dynamics in a patient with arrythmia in order to elicit additional diagnostic information on microvascular dysfunction. METHODS: A 68-year-old male patient with arrythmia and an age- and gender-matched control subject underwent ocular examination including dynamic retinal vessel assessment with flicker light provocation. Retinal vessel diameters were measured continuously following a standard protocol (IMEDOS Systems, Jena, Germany). The data were evaluated using methods of signal analysis. RESULTS: Retinal vessel response following flicker provocation as well as local structural and functional behavior of retinal vessels were comparable between both individuals. The arrhythmia case demonstrated irregular arterial and venous heart rate (HR) pulsation with an average frequency of 1 Hz. Moreover, the case showed a higher magnitude and larger periods of low-frequency retinal vessel oscillations as well as lower periodicity of both HR pulsations and low-frequency vasomotions. CONCLUSIONS: Besides numerical examination of irregular HR pulsations in case of arrhythmia, from the direct noninvasive assessment of retinal vessel dynamics one can derive more detailed information on microvascular function including the whole spectrum of retinal arterial and venous pulsations and vasomotions. This may have implications for health screening not limited to atrial fibrillation.

The Differential Contribution of Macular Pigments and Foveal Anatomy to the Perception of Maxwell's Spot and Haidinger's Brushes.

The relationship of macular pigments and foveal anatomy to the perception of Maxwell's spot (MS) and Haidinger's brushes (HB) entoptic phenomena were investigated. Dual-wavelength-autofluorescence and OCT were used to define macular pigment density and foveal anatomy in 52 eyes. MS was generated by alternating unpolarized red/blue and red/green uniform field illumination. HB was generated by alternating the linear polarization axis of a uniform blue field. In Experiment 1, horizontal widths of MS and HB were measured using a micrometer system and compared with macular pigment densities and OCT-defined morphometry. MS radius (mean 1.4°) was significantly less than HB radius (mean 1.6°), with the spatial extent of both phenomena falling between the boundaries of the foveola and foveal pit. Multiple regression showed MS and HB radii to be significantly associated with the macular pigment spatial profile radius. HB radius, but not MS radius, was also significantly associated with foveolar morphometry. Experiment 2 compared perceptual profiles of MS with macular pigment distribution patterns and demonstrated close agreement. The size and appearance of MS is a direct indicator of macular pigment density and distribution. Measures of HB radii are less specific, with dependence on both macular pigment density and foveal structure.

Dual wavelength retinal vessel oximetry - influence of fundus pigmentation.

BACKGROUND: Clinical methods examining oxygenation parameters in humans have been used in many different care settings, but concerns have been raised regarding their clinical utility when assessing people with darker skin pigmentation. While saturation values can be crucial in emergency medicine, they are equally valuable in assessing disease mechanisms and monitoring change in disease progression. Retinal pigmentation varies across individuals and hence, can impact on retinal oxygen parameters. The objective of this study was to quantify and eliminate the impact of retinal pigmentation on retinal vessel oxygen saturation parameters measured in the superficial retinal arterioles and venules. METHODS: 105 healthy individuals of varying skin colour, iris colour and heritage were included. Following a full eye exam to exclude any ocular abnormality, all participants underwent intraocular pressure, systemic blood pressure measurements and dilated dual wavelength retinal photography. Rotation matrices were employed to minimise the dependency of retinal pigmentation on arterial and venous oxygen saturation measurements determined in a concentric measurement annulus. RESULTS: Retinal oxygen saturation in venules showed a linear correlation with retinal pigmentation (y = 0.34 × x + 38.598), whereas arterial saturation followed a polynomial pattern (y = 0.0089 × x2 + 0.7499 × x + 85.073). Both arterial and venous saturation values were corrected using local fundus pigmentation. Pre-correction retinal arterial and venous oxygen saturation were 89.0% (±13.1) and 43.7% (±11.5), respectively, and post- correction values were 94.8% (±8.7) for arteries and 56.3% (±7.0) veins. CONCLUSIONS: When assessing multi-ethnic cohorts, it is important to consider the impact of pigmentation on imaging parameters and to account for it prior to clinical interpretation.

Size and position of the optic disc crescent in a white European population with myopia.

SIGNIFICANCE: One of the first clinically observed changes in the retina with progressing myopia is in the form of optic disc crescents. If such a change is predictive of myopia progression, it could aid in myopia management interventions to target those at greatest risk of progression and subsequent ocular morbidity. PURPOSE: To investigate the type, dimension and appearance of optic disc crescents and how they relate to the level of myopia. METHODS: Retrospective data collection analysing retinal photographs of healthy children and adults with a refractive error of ≤-0.50 D sphere and astigmatism ≤2.00 D. Crescent location, maximum crescent width and vertical disc diameter were measured from retinal images of right eyes only. RESULTS: Four-hundred eyes with a mean spherical error (SER) of -0.50 to -14.00 D (aged 7-81 years) were included (83.5% exhibited a discernible crescent). Mean (SD) maximum crescent width was 0.24 (0.24) mm. Univariate analysis showed a significant correlation between crescent width and age (r = 0.26, p < 0.001). SER was correlated with crescent width when controlling for age (r = -0.45, p < 0.001) and to the ratio of crescent width to vertical disc diameter (r = -0.43, p < 0.001). Temporal crescents were the most frequently observed (74%), followed by inferior temporal crescents (17%). One-way between-groups analysis of variance showed a significant difference between crescent locations (F = 5.2, p < 0.001). Post-hoc analysis revealed significant differences in SER between those with no crescent versus an inferior-temporal crescent, as well as differences between those with temporal versus inferior-temporal crescents. Other crescent locations did not differ significantly in the level of myopia. Participants not exhibiting a crescent had the lowest level of myopia (mean [SD] -3.03 [1.97)] D), while those with inferior temporal crescents had a mean (SD) SER of -5.01 (2.37) D. CONCLUSION: In this white European population, higher levels of SER were associated with increasing crescent size. Eyes with inferior temporally located crescents were more myopic.

The application of arterio-venous ratio (AVR) cut-off values in clinic to stratify cardiovascular risk in patients.

INTRODUCTION: Cardiovascular risk calculators are a useful tool for identifying at-risk individuals. There are standardised methods for assessing the retinal microcirculation which alters as a consequence of cardiovascular disease (CVD). This study aimed to explore if a standardised retinal vessel assessment conducted in primary optometric care reflects current cardiovascular risk, as measured using two validated CVD risk calculators (QRISK 2; Mayo Clinic). METHODS: A total of 120 subjects were included in the analyses. Following a routine eye examination, participants had disc-centred retinal photographs and systemic blood pressure taken. Retinal vessel parameters (central retinal artery and vein equivalent and arterio-venous ratio (AVR)) were calculated using semi-automated software. Participants were then grouped into AVR quintiles as defined by the Atherosclerosis Risk in Communities Study (ARIC). Cardiovascular risk was calculated with the validated QRISK and Mayo Clinic health calculators. RESULTS: Systolic blood pressure was significantly greater in those with an AVR value falling in the lowest quintile compared to the highest quintile (150.65 mmHg vs. 132.21 mmHg [p = 0.001]). Similarly, CVD risk was significantly higher in those with the lowest AVR compared to the highest (QRISK: 14.28% vs. 9.87% [p = 0.05]; MAYO risk: 36.35% vs. 19.21% [p = 0.01]). Chi squared analyses showed a significant difference in the number of hypertensives in the lowest AVR quintile compared to those in the highest [p = 0.02]. CONCLUSION: Whilst the ARIC population is not directly comparable to the population used to develop the QRISK calculator, it has been shown that its application could help to identify at risk individuals using retinal vessel analyses.

Effect of obstructive sleep apnoea on retinal microvascular function: a randomised controlled trial.

PURPOSE: Retinal microvascular endothelial dysfunction is thought to be of importance in the development of ocular vascular diseases. Obstructive sleep apnoea (OSA) causes macrovascular endothelial dysfunction, but the effect of OSA on retinal microvascular endothelial function is not known. We aimed to determine the effect of OSA on retinal microvascular function. METHODS: We conducted a multi-centre, double-blind, randomised, parallel, controlled trial in patients with known moderate-to-severe OSA, established on continuous positive airway pressure (CPAP). Participants were randomised to 14 nights of either continued CPAP or sham CPAP to generate a return of OSA. Retinal vascular responses to flickering light were measured using dynamic vessel analysis both at baseline and after 14 nights of intervention. The primary outcome was the change from baseline to follow-up in the area under the curve of the arteriolar response to flickering light, sham CPAP versus continued CPAP. RESULTS: Nineteen patients were randomised to sham CPAP, and 18 patients were randomised to continued CPAP. There was no significant effect of CPAP withdrawal and return of OSA on retinal responses, with a change in the area under the curve of the arteriole response to flickering light of + 3.8 arbitrary units (95% CI - 10.6 to + 18.2, p = 0.59), sham CPAP versus continued CPAP. CONCLUSIONS: CPAP withdrawal and a return of OSA had no significant effect on retinal microvascular responses. This contrasts with the effect of CPAP withdrawal on macrovascular endothelial function and suggests that OSA has different effects on macrovascular and microvascular endothelial function. ISRCTN 78082983, 23/10/2014, Prospectively registered.

Comparison of Static Retinal Vessel Caliber Measurements by Different Commercially Available Platforms.

SIGNIFICANCE: Commercially available platforms show good agreement in clinical outcomes for retinal vessel caliber measurements, despite differing absolute values. Tighter agreement is observed when right and left eye data are averaged, suggesting an approach suitable for clinical practice. PURPOSE: The purpose of this study was to compare the retinal vessel caliber measurements generated by different commercially available platforms and their associations with systemic blood pressure and age. METHODS: A total of 209 participants were recruited from a UK optometric practice. After a routine eye examination, participants had disc-centered retinal photographs and systemic blood pressure taken. Retinal vessel calibers (central retinal artery equivalent, central retinal vein equivalent, and arteriovenous ratio) were calculated using both MONA REVA and VesselMap. RESULTS: An inverse Pearson correlation was observed between central retinal artery equivalent and mean arterial blood pressure on both platforms (r = -0.275 [P ≤ .001] and r = -0.388 [P ≤ .001] for MONA REVA and VesselMap, respectively); this correlation was also observed with arteriovenous ratio and blood pressure. An inverse correlation was observed between central retinal artery equivalent and age (r = -0.362 [P ≤ .001] and r = -0.404 [P ≤ .001] for MONA REVA and VesselMap, respectively); this was also seen between central retinal vein equivalent and age (r = -0.322 [P ≤ .001] and r = -0.369 [P ≤ .001]). Arteriovenous ratio remained independent from age for both platforms. Bland-Altman plots demonstrated good agreement between the platforms for all three variables. CONCLUSIONS: Although absolute caliber measurements differed between the platforms, the correlations observed were of similar magnitudes, with good agreement between the two platforms. Tighter spaced limits of agreement were observed when right and left eye data were averaged for each subject. In the absence of localized ocular pathology, this approach should be used.

Comparison between two fast threshold strategies: SPARK and SITA in normal subjects.

BACKGROUND: Numerous fast threshold strategies have been developed in perimetry which use maximum likelihood approaches to estimate the threshold. A recent approach to threshold estimation has been developed estimating the threshold from a limited number of test points which further reduces examination time. This strategy, SPARK, has not been compared to the SITA strategy. The aim of this study was to compare SPARK with SITA in a normal cohort to evaluate within and between strategy agreement in threshold estimates. METHODS: A total of 83 normal subjects each underwent two visual field examinations with SITA and SPARK on two separate occasions on a randomly selected eye. The eye examined and the order of strategy examined first was randomised but remained constant over the two perimetry visits. RESULTS: Visual field examination with SPARK Precision was on average 33% faster than SITA Standard. A positive correlation between group mean sensitivities of SITA Standard and SPARK Precision (rho = 0.713, p < 0.001) was found. In total, 95% of stimulus locations were located within the 95% limits of agreement and linear regression on the differences in sensitivities showed no statistically significant proportional bias (t = 1.713, p = 0.09). Pointwise analysis showed SITA Standard had significantly larger variability for individual stimulus locations examined over two visits when compared to SPARK (t = 9.175, p < 0.001). CONCLUSION: The clinical examination of SPARK yields a sensitivity profile similar to SITA but in a faster examination time. The lower threshold variability of SPARK may be as a result of data smoothing in the threshold estimation process.

Retinal Vessel Oxygen Saturation Measurement Protocols and Their Agreement.

Purpose: To assess agreement between different image sizes and analysis protocols for determination of retinal vessel oxygen saturation in the peripapillary retina of healthy individuals. Methods: Retinal oximetry measurements were acquired from 87 healthy volunteers using the IMEDOS Systems oxygen module. The peripapillary retinal vessels were assessed in a concentric annulus around the optic nerve head. Single and average vessel comparisons were made at different image field sizes of 30° and 50°. Comparisons between images obtained at 30° and 50° were made in a subset of 47 of the 87 individuals. Results: All subjects were normotensive and had normal intraocular pressures (9-16 mm Hg). Analyses of agreement between single vessel, averaged vessel, and between different size images were sought by Bland-Altman analyses, of which all yielded a low bias (<1% oxygen saturation). However, agreement between single vessels of consecutive images showed increased limits of agreement compared with saturation values calculated by averaging all or just the four major arcades of one image. Agreement between 30° and 50° images showed a similar bias as when comparing data obtained with the same camera angle setting but exhibited larger confidence intervals (arteries: bias = 0.21% [9.04/-8.62]%; veins: bias = 0.71% [14.82/-13.40]%). Conclusions: Averaging methods yielded the best agreement; there was little difference in average arterial and venous oxygen saturation between protocols, which analyze all vessels versus the four largest vessels. The least agreement was found for single vessel measurements and comparisons between different camera angles. Translational Relevance: Standardization of image capture protocols (same image size and undertaking a vessel averaging approach for oxygenation analysis) will enhance the detection of smaller physiological changes in eye disease.

Saffron (Crocus sativus L.) in Ocular Diseases: A Narrative Review of the Existing Evidence from Clinical Studies.

Saffron (Crocus sativus L.) and its main constituents, i.e., crocin and crocetin, are natural carotenoid compounds, which have been reported to possess a wide spectrum of properties and induce pleiotropic anti-inflammatory, anti-oxidative, and neuroprotective effects. An increasing number of experimental, animal, and human studies have investigated the effects and mechanistic pathways of these compounds in order to assess their potential therapeutic use in ocular diseases (e.g., in age related macular degeneration, glaucoma, and diabetic maculopathy). This narrative review presents the key findings of published clinical studies that examined the effects of saffron and/or its constituents in the context of ocular disease, as well as an overview of the proposed underlying mechanisms mediating these effects.

Regulation of oxygen saturation in retinal blood vessels in response to dynamic exercise.

PURPOSE: To evaluate the impact of dynamic exercise on retinal vessel oxygen saturation in healthy individuals. METHODS: Twenty-six healthy participants underwent moderate dynamic exercise (modified Master's two-step exercise). In all subjects, intraocular pressures (IOP), systolic and diastolic blood pressures (SBP and DBP), retinal vessel calibres and retinal arterial and venous oxygen saturation were measured at baseline, immediately following exercise and 15 min postexercise. RESULTS: Moderate dynamic exercise increased SBP and DBP immediately postexercise (SBP: 116 (±13) mmHg to 150 (±21) mmHg; p < 0.001 and DBP: 69 (±10) mmHg to 74 (±10); p < 0.001), while IOP decreased by an average of 2 mmHg (baseline: 13 (±3) mmHg)) immediately postexercise (11 (±2) mmHg). Oxygen saturation in retinal arteries remained unchanged (baseline = 93 ± 8%; immediately postexercise = 94 ± 9% and 15 min postexercise = 96 ± 8%; p = 0.069), but increased in retinal veins immediately postexercise and did not return to baseline values within 15 min postexercise (baseline = 54 ± 12%; immediately postexercise = 56 ± 15%; 15 min postexercise=57 ± 12%; p = 0.036). CONCLUSION: There is a mild increase in retinal venous oxygen saturation and there is a trend towards an increase in arterial saturation in otherwise healthy individuals following dynamic exercise.

The influence of simulated cataract on retinal vessel oximetry measurements.

PURPOSE: To assess the impact of human crystalline lens opacification and yellowing, similar to that observed in patients with cataracts, on retinal vessel blood oxygen saturation measurements using custom manufactured soft contact lenses. METHODS: Ten healthy, non-smoking individuals were enrolled for this study. All subjects underwent digital blood pressure measurements, assessment of non-contact intra-ocular pressure, pupil dilation and retinal vessel oximetry using dual-wavelength photography (Oximetry Module, Imedos Systems). To simulate lens changes, three different contact lenses were inserted, one to simulate opacities followed by two more lenses to simulate different levels of lens yellowing (Cantor & Nissel). RESULTS: The measurements obtained showed an opposite change in arterial and venous oxygen saturation and optical density ratio across conditions, resulting in a statistically significant difference in arterial minus venous oxygen saturation value (p = 0.003). However, this difference was only significant for the 'opacity' condition but not for the 'yellowing' conditions. CONCLUSION: Lenticular changes such as cataracts can impact on spectrophotometric analysis in particular dual-wavelength retinal vessel oximetry. Hence, lenticular assessment and cataract grading should be considered when assessing elderly individuals and patient groups developing cataract earlier in life such as those suffering from diabetes mellitus.

The Time Course of Changes in Retinal Vessel Diameter in Response to Differing Durations of Flicker Light Provocation.

PURPOSE: To determine the response of retinal vessels to differing durations of flicker light (FL) stimulation. METHODS: We recorded retinal arterial and venous vessel dilation to 12.5 Hz FL provocation of varying duration (5, 7, 10, and 20 seconds) in 12 healthy young individuals (age range, 26-45 years). All participants underwent a full ocular examination including IOP and blood pressure measurements. RESULTS: Maximum dilation (MD) did not show a significant dependence on flicker duration in arteries, whereas maximum constriction (MC) did. In veins, however, MD significantly increased with flicker duration. Approximately 80% to 90% of MD in arteries is reached within 10 seconds of FL stimulation. CONCLUSIONS: The vast majority of arterial dilatory capacity is reached within 10 seconds of FL stimulation even though venous dilation continues strongly. Since MC of arteries shows a significant dependence on flicker duration, measurements at two different durations can provide more information about the retinal vascular system than at a single flicker duration alone.

Reliability of retinal vessel calibre measurements using a retinal oximeter.

BACKGROUND: Summarised retinal vessel diameters are linked to systemic vascular pathology. Monochromatic images provide best contrast to measure vessel calibres. However, when obtaining images with a dual wavelength oximeter the red-free image can be extracted as the green channel information only which in turn will reduce the number of photographs taken at a given time. This will reduce patient exposure to the camera flash and could provide sufficient quality images to reliably measure vessel calibres. METHODS: We obtained retinal images of one eye of 45 healthy participants. Central retinal arteriolar and central retinal venular equivalents (CRAE and CRVE, respectively) were measured using semi-automated software from two monochromatic images: one taken with a red-free filter and one extracted from the green channel of a dual wavelength oximetry image. RESULTS: Participants were aged between 21 and 62 years, all were normotensive (SBP: 115 (12) mmHg; DBP: 72 (10) mmHg) and had normal intra-ocular pressures (12 (3) mmHg). Bland-Altman analysis revealed good agreement of CRAE and CRVE as obtained from both images (mean bias CRAE = 0.88; CRVE = 2.82). CONCLUSIONS: Summarised retinal vessel calibre measurements obtained from oximetry images are in good agreement to those obtained using red-free photographs.

Comparison of subjective and objective methods to determine the retinal arterio-venous ratio using fundus photography / Comparación de métodos subjetivos y objetivos para determinar el ratio arterio-venoso de la retina, utilizando la fotografía del fondo del ojo

Purpose: To assess the inter and intra observer variability of subjective grading of the retinal arterio-venous ratio (AVR) using a visual grading and to compare the subjectively derived grades to an objective method using a semi-automated computer program. Methods: Following intraocular pressure and blood pressure measurements all subjects underwent dilated fundus photography. 86 monochromatic retinal images with the optic nerve head centred (52 healthy volunteers) were obtained using a Zeiss FF450+ fundus camera. Arterio-venous ratios (AVR), central retinal artery equivalent (CRAE) and central retinal vein equivalent (CRVE) were calculated on three separate occasions by one single observer semi-automatically using the software VesselMap (ImedosSystems, Jena, Germany). Following the automated grading, three examiners graded the AVR visually on three separate occasions in order to assess their agreement. Results: Reproducibility of the semi-automatic parameters was excellent (ICCs: 0.97 (CRAE); 0.985 (CRVE) and 0.952 (AVR)). However, visual grading of AVR showed inter grader differences as well as discrepancies between subjectively derived and objectively calculated AVR (all p<0.000001). Conclusion: Grader education and experience leads to inter-grader differences but more importantly, subjective grading is not capable to pick up subtle differences across healthy individuals and does not represent true AVR when compared with an objective assessment method. Technology advancements mean we no longer rely on opthalmoscopic evaluation but can capture and store fundus images with retinal cameras, enabling us to measure vessel calibre more accurately compared to visual estimation; hence it should be integrated in optometric practise for improved accuracy and reliability of clinical assessments of retinal vessel calibres (AU)

Objetivo: Evaluar la variabilidad de la graduación subjetiva del ratio arterio-vernoso de la retina entre distintos observadores, utilizando la graduación visual, y comparar los grados subjetivamente calculados con un método objetivo que utiliza un programa informático semi-automatizado. Métodos: Tras medir la presión intraocular y la presión sanguínea, se realizó una fotografía del fondo del ojo dilatado a todos los sujetos. Se obtuvieron 86 imágenes monocromáticas de la retina con la cabeza del nervio óptico centrada (52 voluntarios sanos), utilizando una cámara Zeiss FF450+ para el fondo del ojo. Se calcularon los ratios arterio-venosos (AVR), el equivalente de la arteria retiniana central (CRAE) y el equivalente de la vena retiniana central (CRVE) en tres ocasiones diferentes y mediante un único observador, utilizando el software VesselMap (ImedosSystems, Jena, Alemania). Tras la graduación automática, tres examinadores graduaron los AVR visualmente en tres ocasiones diferentes, a fin de valorar la concordancia. Resultados: La reproducibilidad de los parámetros semi-automáticos fue excelente (ICCs: 0,97 (CRAE); 0,985 (CRVE) y 0,952 (AVR)). Sin embargo, la graduación visual del AVR mostró diferencias entre los distintos graduadores, así como discrepancias entre los AVR subjetiva y objetivamente calculados (todas las comparaciones: p<0,000001). Conclusión: La formación y experiencia del graduador origina diferencias entre los diferentes graduadores pero lo más relevante es que la graduación subjetiva no es capaz de recoger las diferencias sutiles entre los sujetos sanos, y no representa el AVR real cuando se compara con un método de valoración objetivo. Los avances tecnológicos implican que no debemos seguir dependiendo de la valoración oftalmoscópica, sino captar y almacenar las imágenes del fondo del ojo tomadas con cámaras retinianas, lo que nos permite medir más precisamente el calibre de los vasos en comparación a la estimación visual; por tanto, dichos avances deberían integrarse en la práctica optométrica para mejorar la precisión y fiabilidad de las valoraciones clínicas de los calibres de los vasos de la retina (AU)

Comparison of subjective and objective methods to determine the retinal arterio-venous ratio using fundus photography.

PURPOSE: To assess the inter and intra observer variability of subjective grading of the retinal arterio-venous ratio (AVR) using a visual grading and to compare the subjectively derived grades to an objective method using a semi-automated computer program. METHODS: Following intraocular pressure and blood pressure measurements all subjects underwent dilated fundus photography. 86 monochromatic retinal images with the optic nerve head centred (52 healthy volunteers) were obtained using a Zeiss FF450+ fundus camera. Arterio-venous ratios (AVR), central retinal artery equivalent (CRAE) and central retinal vein equivalent (CRVE) were calculated on three separate occasions by one single observer semi-automatically using the software VesselMap (ImedosSystems, Jena, Germany). Following the automated grading, three examiners graded the AVR visually on three separate occasions in order to assess their agreement. RESULTS: Reproducibility of the semi-automatic parameters was excellent (ICCs: 0.97 (CRAE); 0.985 (CRVE) and 0.952 (AVR)). However, visual grading of AVR showed inter grader differences as well as discrepancies between subjectively derived and objectively calculated AVR (all p<0.000001). CONCLUSION: Grader education and experience leads to inter-grader differences but more importantly, subjective grading is not capable to pick up subtle differences across healthy individuals and does not represent true AVR when compared with an objective assessment method. Technology advancements mean we no longer rely on opthalmoscopic evaluation but can capture and store fundus images with retinal cameras, enabling us to measure vessel calibre more accurately compared to visual estimation; hence it should be integrated in optometric practise for improved accuracy and reliability of clinical assessments of retinal vessel calibres.