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Photogrammetric analysis of features in human ocular fundus images is affected by various sources of errors, for example aberrations of the camera and eye optics. Another--usually disregarded--type of distortion arises from projecting the near spherical shape of the fundus onto a planar imaging device. In this paper we quantify such projection errors based on geometrical analysis of the reduced model eye imaged by a pinhole camera. The projection error found near the edge of a 50 degrees fundus image is 24%. In addition, the influence of axial ametropia is investigated for both myopia and hyperopia. The projection errors found in hyperopia are similar to those in emmetropia, but decrease in myopia. Spherical as well as ellipsoidal eye shapes were used in the above calculation and their effect was compared. Our results suggest that the simple spherical eye shape is sufficient for correcting projection distortions within a range of ametropia from -5 to +5 diopters.

Laser Doppler measurements performed immediately after the transition from dark adaptation (DA) to light led to the hypothesis that retinal blood flow, Fret, is increased during DA, but the use of visible lasers had prevented measurements during DA. Our aim was to test this hypothesis by measuring Fret during and after DA.Fret in retinal vessels at the optic disk surface was recorded quasi-continuously in one eye of 6 normal subjects (age 27 to 60 years) using a laser Doppler flowmeter in the near-infrared (810 nm). Measurements were performed during light (baseline), various periods of DA and again during light. DA lasted between 2 and 32 min.Average Fret for the 6 subjects did not change significantly (-2.7 +/- 8% sd, p0.05) during the various periods of DA, as determined from linear regressions of the flux versus time. Following the transition from DA to light, there was, in most cases, a rapid transient increase of the flux, which reached an average value of 37 +/- 10% above the pre-transition value and peaked at 30-60 sec after the transition.These results do not support the hypothesis that Fret in normal volunteers is increased during DA. Rather, they strongly suggest that the transient increase in flux observed after DA is induced by the transition from dark to light (FNSRS #3200-043157 et CNR, It. #95.01715.CT04).

To determine whether iris blood flow (IBF) is regulated in response to an acute decrease in mean ocular perfusion pressure (PPm = MOAP-IOP, MOAP = mean ophthalmic arterial pressure) induced by increasing the intraocular pressure (IOP).Iris blood flow was measured using a slit lamp incorporating a laser Doppler flowmetry (LDF) module. The study was conducted on 12 normal volunteers (14 to 59 years old). IOP was raised using a scleral suction cup. In Exp. #1, the suction pressure was successively raised in steps of 50 to 100 mm Hg, each lasting about 10 sec, until IOP reached the MOAP level. In Exp. #2, the suction was raised to 200 mm Hg in 4 successive steps of 2 min duration.In Exp. #1, no significant change of IBF was observed for small decreases of PPm (23%); greater decreases of PPm resulted in a linear IBF decrease (p0.01). In Exp. #2, such a IBF versus PPm decrease was also observed (p0.001). Immediately after release of suction, a significant, transient IBF increase of 79% above baseline level was observed.These results suggest that some IBF regulation occurs for small PPm decreases (23%); no IBF compensatory mechanism appears to operate for further decreases of PPm (23%).

Previous studies have demonstrated, in the cat, a vasodilatation of retinal vessels in response to neuronal activity induced by diffuse luminance flicker. The aim of this study was to determine whether a similar diameter variation is detectable in humans.Nine normal subjects were exposed to 1 min of sinusoidally varying diffuse luminance flicker (10 Hz, 30 degrees around optic nerve head). Monochromatic fundus pictures before and after the stimulation were taken. The diameter of retinal arteries and veins was measured on the digitised photographs with the NIH-Image software and an own algorythm.The diameter immediately after flicker was significantly larger than the pre-stimulus diameter by 4.2 +/- 2.2% (p0.014) (mean +/- SD) for the retinal arteries and 2.7 +/- 1.7% (p0.001) for the retinal veins. Six seconds after cessation of the flicker, arterial diameter was not significantly different from that of pre-flicker value.Diffuse luminance flicker induces an increase in retinal vessel diameter. This suggest that retinal blood flow is coupled with neuronal activity as previously evidenced by the blue field simulation technique in the macula.

Autoregulation of optic nerve head blood flow (Fonh) in response to decreases in perfusion pressure has been demonstrated in animals and humans. The aim of this study was to determine change in Fonh when systemic blood pressure is increased.Blood flow parameters, i.e. relative mean velocity, number, and flux of red blood cells in the ONH tissue (Velonh, Volonh and Fonh, respectively) were measured by laser Doppler flowmetry in one eye of 13 normal subjects (aged 16 to 58 years), at baseline, during, and after isometric exercises consisting of squatting. Brachial artery blood pressure was measured by sphygmomanometry. IOP was measured at baseline and at the end of squatting.During squatting mean arterial pressure increased from 103 +/- 6 mm Hg to 139 +/- 58 mm Hg (average +/- 95% confidence interval), IOP increased from 13 +/- 0.5 to 17 +/- 1 mm Hg. An average increase in PPm from 56 +/- 4 to 80 +/- 7 mm Hg induced no significant (p0.05) change in the blood flow parameters. The sensitivity (detection threshold) of the blood flow changes was 8%.This study shows for the first time in human autoregulation of Fonh when PPm is increased by increasing the systemic blood pressure.

To investigate the response of choroidal blood flow in the foveal region of the human eye to increases in mean perfusion pressure (PPm = mean ophthalmic artery pressure - intraocular pressure; IOP) induced by isometric exercises.Using laser-Doppler flowmetry, changes in velocity (ChBVel), number (ChBVol), and flux (ChBF) of red blood cells in the choroidal vascular system in the foveal region of the fundus were measured in both eyes of 11 normal subjects (ages 18 to 57 years) during isometric exercises.During 90 seconds of squatting, PPm increased by an average of 67%, from 46 to 77 mm Hg. This resulted in a significant increase of 12% in ChBFm (the mean of ChBF during the heart cycle), mainly caused by an increase in ChBVelm. A further increase in PPm to a value approximately 85% above baseline resulted in a 40% increase in ChBFm. A significant negative correlation was found between the changes in ChBVelm and ChBVolm, during squatting.Previous studies have demonstrated that during isometric exercise, blood pressures in the ophthalmic and brachial arteries rise in parallel. These observations and the current results indicate that an increase in PPm up to 67% induces an increase in choroidal vascular resistance that limits the increase in choroidal blood flow to approximately 12%. This regulatory process fails when PPm is further increased.

To investigate the relationship between choroidal blood velocity (ChBVel), blood volume (ChBVol) and blood flow (ChBF) in the foveal region of the human ocular fundus and ocular perfusion pressure and to determine whether the choroidal circulation has some autoregulatory capacity.Measurements of ChBVel, ChBVol and ChBF were obtained by laser Doppler flowmetry in healthy subjects (age range, 21 to 57 years) with normal eye examination results. Measurements were performed at normal intraocular pressure (IOP) and during successive step increases in IOP induced by scleral suction. In experiment 1, in six eyes (five subjects), the IOP was increased rapidly, in steps of 50 to 100 mm Hg of suction pressure, which each lasted approximately 10 seconds to a level above diastolic ophthalmic artery blood pressure (IOP = approximately 72 mm Hg). In experiment 2, in 14 eyes (seven subjects), the IOP was increased slowly in four successive steps at 2-minute intervals to a level of approximately 42 mm Hg. We also determined the pulsatility of the flow parameters during the heart cycle, pulsatility = 1 - diast value/syst value.For both rates of suction cup increase, the relationship between ChBFm (mean ChBF over the heart cycle) and mean perfusion pressure was not linear. At high pressure, ChBFm was less affected by decreases in the pressure than expected from a passive vascular system. In some cases, no change in ChBFm was detectable, although significant changes in PChBF occurred. Further decreases in perfusion pressure resulted in a proportional decrease in ChBFm. On release of suction, a significant increase in ChBFm over baseline value was detectable in experiment 1.The relationship between ChBFm and ocular mean perfusion pressure appears to be bilinear and reveals some autoregulation for moderate step decreases in perfusion pressure. The temporal characteristics of the ChBFm-response suggest a neural or passive hemodynamical process rather than a myogenic or metabolic compensatory mechanism.

Autoregulation is defined as the maintenance of constant blood flow in a vascular system in spite of changes in perfusion pressure (PPm).PPm was decreased by increasing the intraocular pressure (IOP) with a suction cup and optic nerve blood flow was measured with the laser Doppler flowmetry technique (LDF) in 9 normal volunteers.The blood flow was autoregulated down to a PPm of 13 mm Hg (IOP = 47 mm Hg).These results confirm previous studies in cats and monkeys. The mechanism of autoregulation is probably a decrease in resistance due to capillary recruitment.

The effect of changes in ocular perfusion pressure (PPm) on the choroidal blood flow (ChBF) in man was studied with the laser Doppler flowmetry (LDF) technique.We changed the PPm by increasing the intraocular pressure (IOP) or by increasing the blood pressure (BP) with isometric exercises.We observed that a) ChBF was not significantly different from baseline up to an IOP of 27 mm Hg and b) ChBF remained constant even if PPm increased by as much as 72%.Our results suggest that ChBF is autoregulated in response to an increase in IOP up to about 27 mm Hg. ChBF remains also constant in spite of an increase in systemic BP, probably due to a vasoconstriction induced by increased sympathetic activity.

In this paper we present a miniaturized blue field simulator (BFS) which allows the measurement of the velocity (V) pulsatility (P) and number (D) of leukocytes in the macular retinal capillaries. A study on the effect of the aperture size of the blue light source at the subject's pupil on the measured flow parameters was performed.A blue field entoptoscope with a small halogen lamp was used to induce the perception of the "flying corpuscles" and a flat color screen was selected to display a computer simulation of this entoptic phenomenon. The aperture of the blue light source at the pupil was varied by a diaphragm placed at a conjugate pupil plane while the size and retinal irradiance of the blue field stimulus at the fundus was held constant.The results show significant correlations between log pupil area and both V (-18% per log unit) and D (+42% per log unit). When the retinal illuminance is expressed in Log Trolands, V shows no dependency, but D is strongly correlated (+40% per log Troland).When applying the BFS technique, the size of the blue light stimulus at the pupil, if undilated, must be well controlled to minimize instrument-related variations of the blood flow measurements.

To determine the effect of intravascular adenosine on blood flow in the ocular fundus and to examine indirectly whether the blood-brain barrier to adenosine, which exists in the cerebrovasculature of the cat, is present in the eye of this animal.The noninvasive techniques of laser Doppler flowmetry and velocimetry along with fundus photography were used to measure the change in optic nerve head and choroidal and retinal blood flow during intravenous infusions of 0.18 and 0.6 mg/kg per minute of adenosine.Infusions of adenosine induced significant increases in choroidal blood flow (60% with 0.6 mg/kg per minute) but not in optic nerve head or retinal blood flows.The lack of effect of intravenously infused adenosine on the optic nerve and retinal circulations is most likely caused by the tight junctions in the vessels of these vascular beds, which prevent adenosine from reaching its receptors. Perivascular adenosine in the choroid most likely accounts for the increase in blood flow in this tissue.

To develop a noninvasive method for the investigation of choroidal blood flow (ChBF) and its regulation in the foveal region of the human ocular fundus.Measurement of ChBF was based on the technique of laser Doppler flowmetry (LDF). Sixteen normal subjects (age range, 20 to 64 years), with normal eye examination results, were asked to fixate on a diode laser beam (wavelength = 811 nm, 60 microW at the cornea) delivered to the undilated eye through a fundus camera. Light scattered by red blood cells in the tissue volume sampled by the incident laser beam was detected at the fundus image plane of the camera by an optical fiber. The diameter of the beam at the fundus of the emmetropic eye was about 300 microns. Relative ChBF was measured in both eyes by analyzing the Doppler signal with commercial skin blood flowmeters. The average pulsatile component of ChBF, ChBFP, was determined over the cardiac cycle, and its value was compared to the average total ChBF, ChBFAV. Responses of ChBF to various physiological stimuli, such as increased blood O2 and CO2 concentrations, rapid increases in intraocular pressure, and valsalva maneuvers, were documented.Significant correlations were obtained between the ChBFAV values measured with both flowmeters (P0.001) and between the ChBFAV values measured in the right and left eye (P0.001). ChBFP represented less than 23% of ChBFAV. ChBFAV was not significantly affected by 5 minutes of breathing 100% oxygen. Raising end-tidal CO2 in one subject from 37 to 59 mm Hg increased ChBFAV by approximately 40%. Acute elevation of the intraocular pressure by suction cup or finger pressure on the globe reduced ChBFAV by as much as 90%. Valsalva maneuvers induced reproducible responses that were very different from those recorded from the skin microcirculation.Although LDF of the choroidal circulation is still at an early stage of development, this noninvasive method appears to provide continuous and sensitive measurements of relative choroidal blood flow in the foveal region of the human fundus. Near-infrared laser diodes enable measurements through undilated pupils. Examples of responses suggest new avenues in the investigation by LDF of the effect of various physiological stimuli, pharmacologic agents, and pathologic processes on the choroidal circulation in man.

To determine the effect of diffuse luminance flicker on the motion of leukocytes in the retinal macular capillaries of normal subjects.Using the blue field simulation technique, subjects were asked to match the motion of simulated leukocytes displayed on a video monitor to that of their own entoptically seen white blood cells (WBCs). The changes in velocity and density of the WBCs were recorded after stimulation with diffuse luminance flicker of various durations (0 to 16 seconds), either immediately or at various delays (2, 4, 8 seconds) after cessation of the stimulus.White blood cell velocity increased as flicker duration increased from 0 to 16 seconds. After cessation of flicker, leukocyte motion decreased to baseline within 15 seconds.The authors' findings suggest a coupling between retinal neural activity and blood flow in the macular region of the retina. The rapidity of both the flicker-induced increase in WBC motion and the disappearance of the effect after flicker cessation resembles the time course of blood flow changes previously observed in the microcirculation of the cat optic nerve.

The effect of intravenously injected nicardipine on retinal and optic nerve head (ONH) blood flow was studied in 27 cats using laser Doppler velocimetry and flowmetry, respectively. A dose of 20 micrograms/kg of nicardipine had little effect on retinal blood flow. A dose of 100 micrograms/kg, however, produced a significant transient decrease in flow. By contrast, both doses produced a significant increase in ONH blood flow despite a significant decrease of the mean arterial blood pressure. Measurements of the partial pressure of oxygen (PO2) with an oxygen-sensitive microelectrode, whose tip was placed in the vitreous just in front of the optic disc, showed a significant increase in the PO2 that paralleled the increase in ONH blood flow. These results demonstrate, for the first time to the authors' knowledge, a pharmacologically induced increase in ONH blood flow and suggest that nicardipine could have a beneficial effect on ONH tissue.

The acute effect of topical epinephrine HCl 2% on macular capillary blood flow was studied in 18 healthy human volunteers using the blue-field simulation technique. This technique provides a method for quantifying the velocity of leukocytes flowing in macular capillaries. Subjects adjusted the velocity of simulated leukocytes on a computer screen to match that of their own entoptically perceived leukocytes before instilling the drug and 2 hr thereafter. An artificial tear solution was instilled into the fellow eye for a control. Epinephrine instillation resulted in a 8% increase in macular leukocyte velocity and presumably blood flow (P less than 0.03, paired student t-test).

The distributions of blood velocity and volumetric flow rate in individual vessels of the normal human retina were determined as a function of vessel diameter. The mean velocity of blood, Vmean, was calculated from the centerline velocity measured by bidirectional laser Doppler velocimetry (LDV). Volumetric flow rate was determined from Vmean and the vessel diameter, D, measured from monochromatic fundus photographs. Diameter of the arteries and veins at the site of the LDV measurements ranged from 39 to 134 microns and 64 to 177 microns, respectively. Flow velocity correlated with D (P less than 0.001 for both arteries and veins). Volumetric flow rate varied with D at a power of 2.76 +/- 0.16 for arteries and 2.84 +/- 0.12 for veins, in close agreement with Murray's law. Calculated from 12 eyes, the average total arterial and venous volumetric flow rates were 33 +/- 9.6 and 34 +/- 6.3 microliters/min, respectively. The good agreement between both flow rates suggests that the technique and the assumptions for calculating flow yield results that satisfy mass conservation. Total arterial and venous volumetric flow rates correlated with total arterial and venous vessel cross-section. Volumetric flow rate in the temporal retina was significantly greater than in the nasal retina, but the difference is likely to be due to the larger area of the temporal retina. No difference in flow rate was observed between the superior and inferior retinal hemispheres. Finally, blood velocity in the major retinal vessels measured under normal experimental conditions appears remarkably constant over short (hours) and long (months) periods of time.

The effect of acute changes in mean retinal perfusion pressure, P (2/3 of mean brachial artery blood pressure minus IOP), on retinal volumetric blood flow rate, Q, was investigated in normal volunteers. Changes in Q were determined from Q = k X Vmax X D2, where Vmax is the center line red blood cell velocity measured from temporal veins by laser Doppler velocimetry, D is the vessel diameter obtained by monochromatic fundus photography, and k is a constant of proportionality. A suction cup was used to induce step changes in IOP and, consequently, in P. The magnitude of the steps ranged from 10-32 mmHg. During the first 30 sec after a step decrease in P, Vmax and Q were significantly smaller than at rest by an amount proportional to the decrease in P. Thereafter, Vmax and Q increased markedly towards their values at rest, although P changed comparatively little during this period of time. Time constant of the corresponding decrease in vascular resistance, R(t) = P(t)/Q(t), was approximately 45 sec. There was no significant change in D during elevated IOP. Removal of the cup induced an immediate step increase in P, Vmax, D, Q, and R. Thereafter, Vmax, D, Q, and R returned to their values at rest (time constant of the change in R was about 30 sec), while P remained nearly constant. The rapid change in vascular resistance following a step decrease and increase in P can be attributed to an active process that attempts to maintain blood flow close to normal, in spite of changes in perfusion pressure (autoregulation).(ABSTRACT TRUNCATED AT 250 WORDS)

Bidirectional laser Doppler velocimetry (LDV) and fundus photography were applied to investigate the effects of light and darkness on retinal blood flow. Blood flow had increased by an average of 67% after 5 min of darkness. This increase persisted for periods of darkness as long as 80 min (the longest period tested). The magnitude of this elevation, its time course, and its neutralization by the breathing of 100% O2 suggest that, in vivo, the retina consumes more O2 in the dark than in light. This effect is, most probably, associated with the maintenance of the photoreceptor dark current. LDV in light and darkness may become a useful probe of retinal function.

The response of the retinal circulation to an acute elevation in systemic blood pressure was studied in three healthy normotensive volunteers using the noninvasive laser Doppler velocimetry (LDV) technique combined with retinal vessel size measurements. Isometric exercise was employed to induce the acute rise in arterial pressure. There was no detectable change in retinal blood flow until the mean brachial artery blood pressure was elevated to an average of 115 mm Hg, which represented an average rise in mean blood pressure of 41% above baseline values. Above this value, blood flow increased along with further increments in blood pressure. The parallel rise between the ophthalmic artery pressure and the brachial artery pressure indicates that the regulation of retinal blood flow observed in the above mentioned pressure range is achieved through an increase in retinal vascular resistance rather than by a mechanism that would act to maintain a constant ocular perfusion pressure.

The acute effect of cigarette smoking on retinal macular blood flow was studied in 14 healthy habitual smokers using the blue field simulation technique. This technique provides a method for quantifying the velocity of leukocytes flowing in one's own macular capillaries. Subjects adjusted the mean velocity of computer simulated leukocytes moving on a CRT screen to match that of their own entoptically perceived leukocytes before, during, and immediately after smoking a cigarette. Smoking resulted in a 12 +/- 5% (SEM) increase in macular leukocyte velocity and presumably blood flow. Possible mechanisms for this increase are discussed.