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The neuroretinal rims of a set of glaucoma patients were measured using digitized stereo photographs, to determine the reproducibility of computerized stereo measurements of the neuroretinal rim. Each rim was measured five times at 18 locations, with measurement error (ME) defined as the mean of standard deviations of each set of measurements. The following ME were determined: (i) inter-sessional variability (n = 27 right and 24 left eyes, at t1 and t2); (ii) inter-assessor variability (n = 9, 2 assessors); and (iii) variability after colour adjustment algorithms were applied (n = 15). The results were as follows: (i) inter-sessional variability was 3.41+/-1.08 for t1 and 3.22+/-0.84 for t2; (ii) there was a significant difference between the two assessors, although the ME was still low; and (iii) there was no significant differences between the ME of unadjusted and adjusted images. With a measurement error of up to 11% of rim width, these results show that lowcost rim measurements can be made using PC-based software.

The purpose of this study was to evaluate recombinant adeno-associated virus (AAV) as an in vivo gene transfer vector for the retina and to explore the possibility of monitoring the expression of green fluorescent protein (GFP) using a noninvasive method. Rats were injected subretinally with rAAV-gfp or rAAV-lacZ. Strong expression of the reporter gene in a circular area surrounding the injection site was observed in retinal whole mounts and tissue sections. Higher magnification revealed that cells demonstrating high levels of green fluorescence were hexagonal in shape, indicating they were retinal pigment epithelium (RPE) cells. Histological observation of retinal sections demonstrated that recombinant AAV specifically transduced RPE cells. Ten animals were injected with rAAV-gfp for longitudinal studies and the fluorescence was monitored by retinal fluorescence photography. The GFP signal was detected in 100% of the animals as early as 2 weeks postinjection and remained present throughout the experimental period of 4 months. After 2 weeks, a gradual increase in the number of transduced cells occurred before reaching maximal levels of GFP expression at 8 weeks. This was followed by a small decrease over 4 weeks before reaching stable expression at 16 weeks. Our results demonstrated that rAAV efficiently transduces rat RPE cells and that retinal fluorescence photography is suitable for monitoring GFP expression. By using this noninvasive technique, we demonstrated that repetitive measurements of GFP expression in vivo in the rAAV-gfp-transduced retina are possible. This study demonstrated that retinal fluorescence photography is a potent tool for studying AAV-mediated gene delivery in the retina.

Objective: To compare the Zeiss retinal camera with the Nidek 3-Dx camera for photographic quality and stereo separation., Subjects: Eleven subjects (22 eyes) were selected from patients referred for optic disc photography., Methods: The subjects were photographed using the Nidek 3-Dx camera for simultaneous stereo photographs, and the 30 degrees field and 15 degrees field settings on the Zeiss retinal camera for sequential stereo photographs. Four ophthalmologists qualitatively scored the photographs on a five-point scale for stereo separation and photographic resolution and sharpness. The results from the four observers were averaged and the Friedman two-way analysis of variance used to analyse the results., Conclusions: It was found that the stereo separation is significantly better using the Nidek camera when compared with both the 30 degrees and 15 degrees Zeiss results. There was no significant difference in photographic quality between the Nidek and Zeiss 30 degrees photographs. It was also found that the Zeiss 30 degrees images had a significantly higher quality than the Zeiss 15 degrees, which contradicted previous results.

The red-free negatives of 53 right eyes (30 normal eyes and 23 glaucomatous eyes) and 51 left eyes (32 normal eyes and 19 glaucomatous eyes) were analysed using two different image densitometry techniques. The first technique measured the density from rectangular sample areas, while the second measured density from sector-shaped sample areas which more closely follow the course of the nerve fibres in the retina. Indices which measured the deviation of the data from a clinically determined normal 'gold standard' were calculated, and were used to determine the optimum sensitivity and specificity in separating normal from glaucomatous eyes. There is a significant difference between the data from the normal and glaucomatous groups of eyes, when measured from the sector sample areas. The relative efficacy of this technique is also shown by the improved values of sensitivity (from 42%-70% to 70%-91%), although specificity remained fairly constant (from 66%-83% to 62%-88%).

Optic disc and retinal nerve fibre layer (RNFL) photography have been an integral part of the routine screening of glaucoma patients, ocular hypertensives and glaucoma suspects in Perth since 1979. During this period 60 degrees, 40 degrees, 30 degrees and 8 degrees angles of view were examined for potential use in RNFL photography. The degree of dilation and media opacity is highly variable in glaucoma patients because of the age group involved and the miotic therapy. It was found that the 30 degree angle of view gave the most consistent results for routine RNFL photography. Ocular hypertensives tended to be of a lower mean age and 40 degrees wide angle photography was possible but required two different photographic techniques and was not continued. Both 60 degrees and high-magnification RNFL photography are currently used only with selected patients for teaching and demonstration. A modified Hasselblad 120 roll film camera was used for high-resolution RNFL photography. The Hasselblad adaptors, high-resolution results and comparisons with wide-angle RNFL photographs are illustrated in this paper.

Retinal nerve fibre layer (RNFL) photography is an established and integral tool in the management of ocular hypertensives and glaucoma suspects, and in assessing the management of glaucoma in an ophthalmic practice. When RNFL negatives are viewed for analysis, high-contrast negatives yield the most information. When positive prints are subsequently requested, the RNFL negatives are difficult to print. The use of an unsharp mask facilitates the transfer of all the high-contrast negative density range to the print. The use of an unsharp mask has been simplified so that it is quick and efficient and can be adapted for high-contrast negatives other than RNFL images.

Fundus photography is now an established tool for showing early changes in the retinal nerve fibre layer (RNFL). RNFL defects may precede glaucomatous visual field defects by up to five years and any improvement in photography should aid RNFL defect analysis. We have found that the use of an unsharp mask has helped to visualise the RNFL and by using readily available darkroom materials the process of unsharp mask preparation has been simplified.

The main object of this research was to develop a reliable method of screening glaucoma suspects and patients for early loss of or changes in the retinal nerve fiber layer (RNFL). This study quantifies the variances due to photography, digitizing, and analysis of red-free photographs of the RNFL. The influence of pupil size, optic disc position and eye movements, film processing, digitizing, and intra- and interphotographic-session and intra- and interoperator variances were established. It was found that pupils needed to be dilated to at least 6 mm, that the optic disc had to be positioned in a standardized area in the negative, that the head of the subject had to remain still during photography, and that film processing and digitizing of the negative needed to be strictly controlled to minimize the variance in collection of densitometry data from RNFL red-free photographs. It was established that focusing of the negatives during digitization was not crucial. Criteria were defined for acceptable negatives. Interphotographic-session and intraoperator variances were not significant in most cases when negatives were digitized to these criteria. Analysis of interphotographic-session variance showed that there were still some factors in photography, film processing, and/or image digitizing that were not sufficiently controlled for long-term follow-up without normalization of the data. Densitometry data gathered using the established protocol, from negatives of 71 subjects were analyzed; best sensitivity and specificity rates of 80% and 100%, respectively, were achieved for the diagnosis of glaucoma.