Mahnaz Shahidi

Thickness profiles of retinal layers by optical coherence tomography image segmentation.

PURPOSE To report an image segmentation algorithm that was developed to provide quantitative thickness measurement of six retinal layers in optical coherence tomography (OCT) images. DESIGN Prospective cross-sectional study. METHODS Imaging was performed with time- and spectral-domain OCT instruments in 15 and 10 normal healthy subjects, respectively. A dedicated software algorithm was developed for boundary detection based on a 2-dimensional edge detection scheme, enhancing edges along the retinal depth while suppressing speckle noise. Automated boundary detection and quantitative thickness measurements derived by the algorithm were compared with measurements obtained from boundaries manually marked by three observers. Thickness profiles for six retinal layers were generated in normal subjects. RESULTS The algorithm identified seven boundaries and measured thickness of six retinal layers: nerve fiber layer, inner plexiform layer and ganglion cell layer, inner nuclear layer, outer plexiform layer, outer nuclear layer and photoreceptor inner segments (ONL+PIS), and photoreceptor outer segments (POS). The root mean squared error between the manual and automatic boundary detection ranged between 4 and 9 mum. The mean absolute values of differences between automated and manual thickness measurements were between 3 and 4 mum, and comparable to interobserver differences. Inner retinal thickness profiles demonstrated minimum thickness at the fovea, corresponding to normal anatomy. The OPL and ONL+PIS thickness profiles respectively displayed a minimum and maximum thickness at the fovea. The POS thickness profile was relatively constant along the scan through the fovea. CONCLUSIONS The application of this image segmentation technique is promising for investigating thickness changes of retinal layers attributable to disease progression and therapeutic intervention.

Higher-order wavefront aberrations in corneal refractive therapy.

Purpose. To assess the changes in higher-order (third through sixth) ocular wavefront aberrations produced by Corneal Refractive Therapy (CRT; Paragon Vision Sciences, Mesa, AZ). Methods. Eighteen eyes of nine myopic subjects were fit with CRT contact lenses. Baseline subjective spherical refraction ranged from −2.25 to −6.00 D (mean ± SD, −3.33 ± 1.26 D), and baseline logarithm of the minimum angle of resolution (logMAR) best-corrected visual acuity was −0.13 ± 0.06 (20/15 Snellen equivalent). Whole-eye ocular wavefront aberrations were measured using a previously validated Shack-Hartmann aberrometer. Measurements were taken at baseline and 1 month after treatment initiation. Nine measurements per dilated subject were taken and averaged. Zernike coefficients were used to calculate the third-, fourth-, fifth-, and sixth-order root-mean-square values for each eye for both 3- and 6-mm pupil sizes, and aberrations were averaged and compared with prior baseline readings. Results. The mean (±SD) myopia reduction was 3.08 ± 0.93 D, resulting in a subjective refraction of −0.22 ± 0.38 D after 1 month of lens wear. Both logMAR uncorrected visual acuity (−0.07 ± 0.18; 20/15− Snellen equivalent) and best-corrected visual acuity (−0.14 ± 0.09) after CRT wear were not significantly different from baseline logMAR best-corrected visual acuity (paired two-tailed t-test; p = 0.41 and 0.65, respectively). Whole-eye aberrations showed a statistically significant increase in higher-order aberrations for both 3-mm (factor of 2.66; p = 0.01) and 6-mm pupils (factor of 2.50; p = 0.005). Each individual higher-order aberration also increased, ranging from a factor of 2.01 to 3.20 for 3-mm pupil sizes and 2.52 to 2.98 for 6-mm pupil sizes. Spherical-like aberrations (S4 and S6) increased by a factor of 1.79 for 3-mm pupil sizes and 2.42 for 6-mm pupil sizes. The Zernike coefficient most affected by CRT was spherical aberration (Z40), which increased from 0.084 ± 0.16 to 0.39 ± 0.16 &mgr;m (p = 0.0002) for 6-mm pupils. Conclusions. Use of current CRT lenses for the reduction of myopia increased higher-order wavefront aberrations and spherical aberration (Z40) in particular.

Differential Macular Morphology in Patients with RPE65-, CEP290-, GUCY2D-, and AIPL1-Related Leber Congenital Amaurosis

PURPOSE To evaluate genotypic and macular morphologic correlations in patients with RPE65-, CEP290-, GUCY2D-, or AIPL1-related Leber congenital amaurosis (LCA) using spectral-domain optical coherence tomography (SD-OCT). METHODS SD-OCT macular scans were performed in 21 patients, including 10 with RPE65, 7 with CEP290, 3 with GUCY2D, and 1 with AIPL1 mutations. An image processing software was used to manually draw segmentation lines by three observers. Lamellar structure was evaluated based on the number of retinal layers on segmented images. Total retinal thickness was measured at the central macular and perifoveal areas by using an automated algorithm. RESULTS All three patients with GUCY2D mutations (age range, 20-53 years) retained six retinal layers with visible photoreceptor inner/outer segment juncture (PSJ). However, the preservation of lamellar structures did not parallel better visual acuity. Patients with other mutations had poorly defined PSJ and disorganized retinal lamellar structures, where only one to three retinal layers could be observed. Patients with CEP290 mutations trended to have retention of the outer nuclear layer at the fovea and macular thickening, especially at younger ages. In patients with RPE65 (age range, 20-71 years) and AIPL1 mutations (age, 22 years), macular thickness was markedly decreased. Disorganization of retinal lamellar structures in the RPE65 group trended toward a worsening with increasing age. CONCLUSIONS Variations of macular microstructures were observed among LCA patients with different genotypes. Disorganization of retinal lamellar structure was generally age related. Preservation of retinal microanatomic structures may not be associated with better visual acuity.

Topography of the Retinal Thickness in Normal Subjects

A noninvasive method has been developed that is capable of providing quantitative thickness profiles of the retina. The method was used to map the retinal thickness in five normal human volunteers and determine the reproducibility of the measurements. The reproducibility or equivalent sensitivity of the measurements to detect changes was found to be 5% or 19 microns on the same day and 8% or 31 microns on different days. By averaging the values obtained in five normal subjects, ranging in age from 21 to 43 years (mean, 34 years), a preliminary normal baseline was derived for the thickness profile at the fovea and the thickness cross-section from the optic disc to the fovea. The results of the study indicated that this noninvasive method promises to be of clinical use in diagnosing ocular diseases that produce changes in the thickness of the retinal as well as in monitoring the effectiveness of therapy.

A Method for Chorioretinal Oxygen Tension Measurement

Purpose: To report an optical imaging system that was developed to measure oxygen tension (pO2) in the chorioretinal vasculatures. The feasibility of the system for the measurement of changes in pO2 separately in the retinal and choroidal vasculatures was established in rat eyes by varying the fraction of inspired oxygen and inhibiting nitric oxide activity. Methods: Our optical section phosphorescence imaging system was modified to provide quantitative measurements of pO2 separately in the retinal and choroidal vasculatures. A narrow laser line was projected at an angle on the retina after intravenous injection of an oxygen-sensitive probe (Pd-porphyrin), and phosphorescence emission was imaged. A frequency-domain approach allowed measurements of the phosphorescence lifetime by varying the phase relationship between the modulated excitation laser light and sensitivity of the imaging camera. Chorioretinal pO2 was measured while varying the fraction of inspired oxygen and during intravenous infusion of Nω-nitro-L-arginine (Nω-NLA), a nonselective nitric oxide synthase inhibitor. Results: The systemic arterial pO2 varied according to the fraction of inspired oxygen. The pO2 in the retinal and choroidal vasculatures increased as the fraction of inspired oxygen was increased. Compared with baseline, choroidal pO2 decreased during infusion of Nω-NLA, whereas the pO2 in the retinal vasculatures remained relatively unchanged. The choroidal pO2 decreased markedly with each incremental increase in Nω-NLA infusion rate, in the range 1–6 mg/min, and there was no additional change in the choroidal pO2 at Nω-NLA infusion rates above 6 mg/min. Conclusions: An optical method combining pO2 phosphorescence imaging with chorioretinal optical sectioning was established that can potentially be applied for better understanding of retinal and choroidal oxygen dynamics in physiologic and pathologic states.

Central Macular Splaying and Outer Retinal Thinning in Asymptomatic Sickle Cell Patients by Spectral Domain Optical Coherence Tomography

PURPOSE To investigate the prevalence and degree of macular thinning on optical coherence tomography (SDOCT) in African-American female patients with asymptomatic sickle cell disease. DESIGN Prospective comparative case series. METHODS Twenty-one sickle cell patients (42 eyes) without other systemic or ocular diseases and 18 healthy control patients (33 eyes) underwent SD-OCT. Images were manually segmented to measure inner retinal thickness (IRT) and outer retinal thickness (ORT). Central macular (central 1 mm), parafoveal (0.5-1.5 mm eccentricity), and perifoveal (1.5-3 mm eccentricity) thickness measurements were obtained in sickle cell patients and age/gender/race-matched healthy control subjects. RESULTS Central macular total thickness (CMT) in sickle cell patients was 220 ± 3 μm (mean ± SEM), which was significantly lower (P < .05) than controls (228 ± 3 μm). Parafoveal regions had thickness measurements of 314 ± 5 μm (nasal) and 304 ± 2 μm (temporal), which were significantly lower than controls (327 ± 2 μm and 311 ± 2 μm nasally and temporally, respectively) (P < .03, P < .043). There was also no significant difference in IRT in central macula, parafoveal, and perifoveal regions. Central macular ORT was 175 ± 2 μm vs 185 ± 1 μm in controls (P < .0002). ORT in temporal parafoveal and perifoveal regions was 142 ± 2 μm and 120 ± 1 μm, respectively, vs 150 ± 1 μm and 122 ± 1 μm in controls (P < .001 and P = .16, respectively). CONCLUSIONS Manual segmentation of SD-OCT images revealed significant total retinal thinning in the central macula and splaying in asymptomatic sickle cell patients. Retinal thinning was predominately in outer retinal layers in central macula and parafoveal regions.

Three-dimensional mapping of chorioretinal vascular oxygen tension in the rat.

PURPOSE An optical section phosphorescence lifetime imaging system was developed for three-dimensional mapping of oxygen tension (P(O2)) 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 P(O2) was measured by determining the phosphorescence lifetime. Three-dimensional chorioretinal P(O2) maps were generated in rat eyes under varying fractions of inspired oxygen. RESULTS Under an air-breathing condition, mean P(O2) 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 P(O2) difference was 12+/-2 mm Hg. With a lower fraction of inspired oxygen, chorioretinal vascular P(O2) and mean arteriovenous P(O2) differences decreased compared with measurements under an air-breathing condition. Retinal venous P(O2) was statistically lower than P(O2) measured in the retinal artery, capillaries, and choroid (P<0.004). CONCLUSIONS Three-dimensional mapping of chorioretinal oxygen tension allowed quantitative P(O2) 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.

Selective thinning of the perifoveal inner retina as an early sign of hydroxychloroquine retinal toxicity.

PurposeTo evaluate macular thickness profiles using spectral-domain optical coherence tomography (SDOCT) and image segmentation in patients with chronic exposure to hydroxychloroquine.MethodsThis study included eight patients with chronic exposure to hydroxychloroquine (group 1) and eight controls (group 2). Group 1 patients had no clinically evident retinal toxicity. All subjects underwent SDOCT imaging of the macula. An image segmentation technique was used to measure thickness of six retinal layers at 200 μm intervals. A mixed-effects model was used for multivariate analysis.ResultsBy measuring total retinal thickness either at the central macular (2800 μm in diameter), the perifoveal region 1200-μm-width ring surrounding the central macula), or the overall macular area (5200 μm in diameter), there were no significant differences in the thickness between groups 1 and 2. On an image segmentation analysis, selective thinning of the inner plexiform+ganglion cell layers (P=0.021) was observed only in the perifoveal area of the patients in group 1 compared with that of group 2 by using the mixed-effects model analysis.ConclusionOur study results suggest that chronic exposure to hydroxychloroquine is associated with thinning of the perifoveal inner retinal layers, especially in the ganglion cell and inner plexiform layers, even in the absence of functional or structural clinical changes involving the photoreceptor or retinal pigment epithelial cell layers. This may be a contributing factor as the reason most patients who have early detectable signs of drug toxicity present with paracentral or pericentral scotomas.

Quantitative assessment of conjunctival microvascular circulation of the human eye

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 was 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.

Clinical Assessment of the Macula by Retinal Topography and Thickness Mapping

PURPOSE To report a quantitative and objective method for assessing pathologic alterations in retinal structures to improve the evaluation of macular diseases. METHODS We used a system based on the scanning retinal thickness analyzer to generate serial optical section images of the retina and provide mapping of the retinal topography and thickness in a normal subject and in patients with representative maculopathies including traumatic macular hole, central serous chorioretinopathy, branch retinal vein occlusion, diabetic macular edema, and retinal pigment epithelial detachment. RESULTS The effectiveness of the system in imaging both the vitreoretinal and chorioretinal interfaces was confirmed in the normal subject and in patients with various maculopathies. Mapping of retinal topography and thickness in a normal eye correlated well with normal anatomy, delineating the foveal depression clearly. The retinal thickness map in a patient with diabetic macular edema showed thickening of the retina and absence of a foveal depression. The patients with central serous chorioretinopathy and branch retinal vein occlusion had an elevated vitreoretinal surface. Conversely, the patient with retinal pigment epithelial detachments had a relatively flat vitreoretinal interface but an irregularly elevated chorioretinal surface. CONCLUSION Quantitative mapping of retinal topography and thickness is a promising tool that may improve evaluation of macular diseases.