Visual pigments, cone oil droplets and ocular media in four species of estrildid finch

A microspectrophotometric study was conducted on the retinal photoreceptors of four species of bird: cut-throat finches (Amadina fasciata), gouldian finches (Erythrura gouldiae), white-headed munias (Lonchura maja) and plum-headed finches (Neochmia modesta). Spectral characteristics of the photoreceptors in all four species were very similar. Rods contained a medium-wavelength-sensitive visual pigment with a wavelength of maximum absorbance at 502-504 nm. Four spectrally distinct types of single cone contained a visual pigment with wavelength of maximum absorbance at either 370-373 nm (ultraviolet-sensitive), 440-447 nm (short-wavelength-sensitive); 500 nm (medium-wavelength-sensitive) or 562-565 nm (long-wavelength-sensitive). Oil droplets in the ultraviolet-sensitive single cones showed no detectable absorption between 330 nm and 800 nm. Oil droplets in the short-, medium-, and long-wavelength-sensitive single cones had cut-off wavelengths at 415-423 nm, 510-520 nm and 567-575 nm, respectively. Double cones contained the visual pigment with wavelength of maximum absorbance at 562-565 nm observed in long-wavelength-sensitive single cones. Only the principal member of the double cone pair contained an oil droplet (P-type, cut-off wavelength at 414-489 nm depending on species and retinal location). Spectral transmittance of the intact ocular media of each species was measured along the optic axis. Wavelengths of 0.5 transmittance for all species were very similar (316-318 nm).

Visual pigments, oil droplets, ocular media and cone photoreceptor distribution in two species of passerine bird: the blue tit (Parus caeruleus L.) and the blackbird (Turdus merula L.)

The spectral absorption characteristics of the retinal photoreceptors of the blue tit (Pal trs caeruleus) and blackbird (Turdus merula) were investigated using microspectrophotometry. The retinae of both species contained rods, double cones and four spectrally distinct types of single cone. Whilst the visual pigments and cone oil droplets in the other receptor types are very similar in both species, the wavelength of maximum sensitivity (lambda(max)) of long-wavelength-sensitive single and double cone visual pigment occurs at a shorter wavelength (557 nm) in the blackbird than in the blue tit (563 nm). Oil droplets located in the long-wavelength-sensitive-single cones of both species cut off wavelengths below 570-573 nm, theoretically shifting cone peak spectral sensitivity some 40 nm towards the long-wavelength end of the spectrum. This raises the possibility that the precise lambda(max) of the long-wavelength-sensitive visual pigment is optimised for the visual function of the double cones. The distribution of cone photoreceptors across the retina, determined using conventional light and fluorescence microscopy also varies between the two species and may reflect differences in their visual ecology.

Refractive indices with Haag-Streit Lenstar biometer

Purpose: To estimate refractive indices used with the Lenstar biometer. Methods: Axial lengths of model eyes were determined using an IOLMaster biometer and a Lenstar; comparing these lengths gave an overall eye index for the Lenstar. Using the Lenstar Graphical User interface, we determined that boundaries between media could be manipulated so that there were opposite changes in optical pathlength on either side of the boundary and specified changes in distances determined the ratios of media indices. These ratios were combined with the overall eye index to estimate indices. Results: The IOLMaster and Lenstar produced axial length estimates to within ±0.01 mm. Estimations of group refractive indices were 1.340, 1.341, 1.415 and 1.354 for cornea, aqueous, lens and overall eye, respectively. The aqueous and lens indices, but not those for the cornea, are similar to schematic eye indices and reasonable lens indices. Conclusion: The Lenstar appears to use different refractive indices for different ocular media.

Refractive indices used by the Haag-Streit Lenstar to calculate axial biometric dimensions

PURPOSE To estimate refractive indices used by the Lenstar biometer to translate measured optical path lengths into geometrical path lengths within the eye. METHODS Axial lengths of model eyes were determined using the IOLMaster and Lenstar biometers; comparing those lengths gave an overall eye refractive index estimate for the Lenstar. Using the Lenstar Graphical User Interface, we noticed that boundaries between media could be manipulated and opposite changes in optical path lengths on either side of the boundary could be introduced. Those ratios were combined with the overall eye refractive index to estimate separate refractive indices. Furthermore, Haag-Streit provided us with a template to obtain 'air thicknesses' to compare with geometrical distances. RESULTS The axial length estimates obtained using the IOLMaster and the Lenstar agreed to within 0.01 mm. Estimates of group refractive indices used in the Lenstar were 1.340, 1.341, 1.415, and 1.354 for cornea, aqueous, lens, and overall eye, respectively. Those refractive indices did not match those of schematic eyes, but were close in the cases of aqueous and lens. Linear equations relating air thicknesses to geometrical thicknesses were consistent with our findings. CONCLUSION The Lenstar uses different refractive indices for different ocular media. Some of the refractive indices, such as that for the cornea, are not physiological; therefore, it is likely that the calibrations in the instrument correspond to instrument-specific corrections and are not the real optical path lengths.

Les dérivées de l’hémoglobine dans la structure rétinienne par la technique de réflectométrie multi-spectrale

L’examen de la rétine par des moyens non invasifs et in vivo a été un objectif de recherche pendant plusieurs années. Pour l’œil comme pour tous les organes du corps humain, un apport soutenu en oxygène est nécessaire pour le maintien de l’homéostasie. La concentration en oxygène du sang des vaisseaux rétiniens peut être déterminée principalement à partir des mesures du spectre de réflexion du fond de l’œil. En envoyant une lumière, à différentes longueurs d’onde, sur la rétine et en analysant la nature de la lumière réfléchie par la rétine, il est possible d’obtenir des informations quantitatives sur le niveau d'oxygène dans les vaisseaux sanguins de la rétine ou sur le flux sanguin. Cependant, la modélisation est compliquée due aux différentes interactions et aux chemins que la lumière prend à travers les tissus oculaires avant de quitter l’œil. L’objectif de cette thèse a été de développer et de valider un modèle mathématique afin de calculer les dérivées d’hémoglobine à partir de mesures spectrales de réflectométrie sur les vaisseaux sanguins de la rétine. L’instrument utilisé pour mesurer la fonction spectrale de réflectométrie a été un spectroréflectomètre multi-canal, une technologie capable de mesurer in vivo et en continu 800 spectres simultanément. L'équation mathématique qui décrit la fonction spectrale de réflectométrie dans la zone spectrale de 480 nm à 650 nm a été exprimée comme la combinaison linéaire de plusieurs termes représentant les signatures spectrales de l'hémoglobine SHb, de l'oxyhémoglobine SOHB, l’absorption et la diffusion des milieux oculaires et une famille de fonctions multigaussiennes utilisées pour compenser l’incompatibilité du modèle et les données expérimentales dans la zone rouge du spectre. Les résultats du modèle révèlent que le signal spectral obtenu à partir de mesures de réflectométrie dans l’œil est complexe, contenant la lumière absorbée, réfléchie et diffusée, mais chacun avec une certaine prédominance spécifique en fonction de la zone spectrale. La fonction spectrale d’absorption du sang est dominante dans la zone spectrale 520 à 580 nm, tandis que dans la zone spectrale de longueurs d’ondes plus grandes que 590 nm, la diffusion sur les cellules rouges du sang est dominante. Le modèle a été utilisé afin de mesurer la concentration d’oxygène dans les capillaires de la tête du nerf optique suite à un effort physique dynamique. L’effort physique a entraîné une réduction de la concentration d’oxygène dans les capillaires, ainsi qu’une réduction de la pression intraoculaire, tandis que la saturation sanguine en oxygène, mesurée au niveau du doigt, restait constante. Le modèle mathématique développé dans ce projet a ainsi permis, avec la technique novatrice de spectroréflectométrie multicanal, de déterminer in vivo et d’une manière non invasive l’oxygénation sanguine des vaisseaux rétiniens.

Ultraviolet colour perception in European starlings and Japanese quail

Whereas humans have three types of cone photoreceptor, birds have four types of single cones and, unlike humans, are sensitive to ultraviolet light (UV, 320-400 run). Most birds are thought to have either a violet-sensitive single cone that has some sensitivity to UV wavelengths (for example, many non-passerine species) or a single cone that has maximum sensitivity to UV (for example, oscine passerine. species). UV sensitivity is possible because, unlike humans, avian ocular media do not absorb UV light before it reaches the retina. The different single cone types and their sensitivity to UV light give birds the potential to discriminate reflectance spectra that look identical to humans. It is clear that birds use UV signals for a number of visual tasks, but there are few studies that directly demonstrate a role for UV in the detection of chromaticity differences (i.e. colour vision) as opposed to achromatic brightness. If the output of the violet/UV cone is used in achromatic visual tasks, objects reflecting more UV will appear brighter to the bird. 11, however, the output is used in a chromatic mechanism, birds will be able to discriminate spectral stimuli according to the amount of reflected light in the UV part of the spectrum relative to longer wavelengths. We have developed a UV 'colour blindness' test, which we have given to a passerine (European starling) and a non-passerine (Japanese quail) species. Both species learnt to discriminate between a longwave control of orange vs red stimuli and UV vs 'non-UV' stimuli, which were designed to be impossible to differentiate by achromatic mechanisms. We therefore conclude that the output of the violet/UV cone is involved in a chromatic colour vision system in these two species.

Propagation, structural similarity and image quality

Póster presentado en SPIE Photonics Europe, Brussels, 16-19 April 2012.

Microspectrophotometry of visual pigments and oil droplets in a marine bird, the wedge-tailed shearwater Puffinus pacificus: Topographic variations in photoreceptor spectral characteristics

Microspectrophotometric examination of the retina of a procellariiform marine bird, the wedge-tailed shearwater Puffinus pacificus, revealed the presence of five different types of vitamin A(1)-based visual pigment in seven different types of photoreceptor. A single class of rod contained a medium-wavelength sensitive visual pigment with a wavelength of maximum absorbance (lambda(max)) at 502 nm. Four different types of single cone contained visual pigments maximally sensitive in either the violet (VS, lambda(max) 406 nm), short (SWS, lambda(max) 450 nm), medium (MWS, lambda(max) 503 nm) or long (LWS, lambda(max) 566 nm) spectral ranges. In the peripheral retina, the SWS, MWS and LWS single cones contained pigmented oil droplets in their inner segments with cut-off wavelengths (lambda(cut)) at 445 (C-type), 506 (Y-type) and 562 nm (R-type), respectively. The VS visual pigment was paired with a transparent (T-type) oil droplet that displayed no significant absorption above at least 370 run. Both the principal and accessory members of the double cone pair contained the same 566 nm lambda(max) visual pigment as the LWS single cones but only the principal member contained an oil droplet, which had a lambda(cut) at 413 nm. The retina had a horizontal band or 'visual streak' of increased photoreceptor density running across the retina approximately 1.5 mm dorsal to the top of the pecten. Cones in the centre of the horizontal streak were smaller and had oil droplets that were either transparent/colourless or much less pigmented than at the periphery. It is proposed that the reduction in cone oil droplet pigmentation in retinal areas associated with high visual acuity is an adaptation to compensate for the reduced photon capture ability of the narrower photoreceptors found there. Measurements of the spectral transmittance of the ocular media reveal that wavelengths down to at least 300 nm would be transmitted to the retina.

Communication in coral reef fish: the role of ultraviolet colour patterns in damselfish territorial behaviour

Many coral reef fish possess ultraviolet (UV) colour patterns. The behavioural significance of these patterns is poorly understood and experiments on this issue have not been reported for free-living reef fish in their natural environment. The damselfish Pomacentrus amboinensis has UV facial patterns, and spectroradiometric ocular media measurements show that it has the potential for UV vision. To test the potential behavioural significance of the UV patterns, I studied the response of males, in natural territories on the reef and in aquaria, to two conspecific intruders, one presented in a UV-transmitting (UV+) container and the other in a UV-absorbing (UV-) one. Territory owners attacked intruders viewed through UV+ filters significantly more often and for longer than intruders viewed through the UV- filter. In general, the results of the field experiment confirmed those of the laboratory experiment. The results support the hypothesis that P. amboinensis males are sensitive to UV light and that reflectance patterns, which appear in high contrast only in UV, modulate the level of aggressive behaviour. A recent survey showed that many predatory fish may not have UV vision and the use of UV colours in select species of reef fish may therefore serve as a 'private communication channel'. (C) 2004 The Association for the Study of Animal Behaviour. Published by Elsevier Ltd. All rights reserved.

Modelling oil droplet absorption spectra and spectral sensitivities of bird cone photoreceptors

Birds have four spectrally distinct types of single cones that they use for colour vision. It is often desirable to be able to model the spectral sensitivities of the different cone types, which vary considerably between species. However, although there are several mathematical models available for describing the spectral absorption of visual pigments, there is no model describing the spectral absorption of the coloured oil droplets found in three of the four single cone types. In this paper, we describe such a model and illustrate its use in estimating the spectral sensitivities of single cones. Furthermore, we show that the spectral locations of the wavelengths of maximum absorbance (lambda(max)) of the short- (SWS), medium- (MWS) and long- (LWS) wavelength-sensitive visual pigments and the cut-off wavelengths (lambda(cut)) of their respective C-, Y- and R-type oil droplets can be predicted from the lambda(max) of the ultraviolet- (UVS)/violet- ( VS) sensitive visual pigment.

Examination variability in short-wavelength automated perimetry

The study evaluated sources of within- and between-subject variability in standard white-on-white (W-W) perimetry and short-wavelength automated perimetry (SWAP). The Influence of staircase strategy on the fatigue effect in W-W perimetry was investigated for a 4 dB single step, single reversal strategy; a variable step size, single reversal dynamic strategy; and the standard 4-2 dB double reversal strategy. The fatigue effect increased as the duration of the examination Increased and was greatest in the second eye for all strategies. The fatigue effect was lowest for the 4dB strategy, which exhibited the shortest examination time and was greatest for the 4-2 dB strategy, which exhibited the longest examination time. Staircase efficiency was lowest for the 4 dB strategy and highest for the dynamic strategy which thus offers a reduced examination time and low inter-subject variability. The normal between-subject variability of SWAP was determined for the standard 4-2 dB double reversal strategy and the 3 dB single reversal FASTPAC strategy and compared to that of W-W perimetry, The decrease in sensitivity with Increase in age was greatest for SWAP. The between-subject variability of SWAP was greater than W-W perimetry. Correction for the Influence of ocular media absorption reduced the between-subject variability of SWAP, The FASTPAC strategy yielded the lowest between-subject variability In SWAP, but the greatest between-subject variability In WoW perimetry. The greater between-subject variability of SWAP has profound Implications for the delineation of visual field abnormality, The fatigue effect for the Full Threshold strategy in SWAP was evaluated with conventional opaque, and translucent occlusion of the fellow eye. SWAP exhibited a greater fatigue effect than W-W perimetry. Translucent occlusion reduced the between-subject variability of W-W perimetry but Increased the between-subject variability of SWAP. The elevation of sensitivity was greater with translucent occlusion which has implications for the statistical analysis of W-W perimetry and SWAP. The influence of age-related cataract extraction and IOL implantation upon the visual field derived by WoW perimetry and SWAP was determined. Cataract yielded a general reduction In sensitivity which was preferentially greater in SWAP, even after the correction of SWAP for the attenuation of the stimulus by the ocular media. There was no correlation between either backward or forward light scatter and the magnitude of the attenuation of W-W or SWAP sensitivity. The post-operative mean deviation in SWAP was positive and has ramifications for the statistical Interpretation of SWAP. Short-wavelength-sensitive pathway isolation was assessed as a function of stimulus eccentricity using the two-colour Increment threshold method. At least 15 dB of SWS pathway Isolation was achieved for 440 nm, 450 nm and 460 nm stimuli at a background luminance of 100 cdm-2, There was a slight decrease In SWS pathway Isolation for all stimulus wavelengths with increasing eccentricity which was not of clinical significance. Adopting a 450 nm stimulus may reduce between-subject variability In SWAP due to a reduction In ocular media absorption and macular pigment absorption.

Investigation of chromatic function in the normal and abnormal eye

This study examined the use of non-standard parameters to investigate the visual field, with particular reference to the detection of glaucomatous visual field loss. Evaluation of the new perimetric strategy for threshold estimation - FASTPAC, demonstrated a reduction in the examination time of normals compared to the standard strategy. Despite an increased within-test variability the FASTPAC strategy produced a similar mean sensitivity to the standard strategy, reducing the effects of patient fatigue. The new technique of Blue-Yellow perimetry was compared to White-White perimetry for the detection of glaucomatous field loss in OHT and POAG. Using a database of normal subjects, confidence limits for normality were constructed to account for the increased between-subject variability with increase in age and eccentricity and for the greater variability of the Blue-Yellow field compared to the White-White field. Effects of individual ocular media absorption had little effect on Blue-Yellow field variability. Total and pattern probability analysis revealed five of 27 OHTs to exhibit Blue-Yellow focal abnormalities; two of these patients subsequently developed White-White loss. Twelve of the 24 POAGs revealed wider and/or deeper Blue-Yellow loss compared with the White-White field. Blue-Yellow perimetry showed good sensitivity and specificity characteristics, however, lack of perimetric experience and the presence of cataract influenced the Blue-Yellow visual field and may confound the interpretation of Blue-Yellow visual field loss. Visual field indices demonstrated a moderate relationship to the structural parameters of the optic nerve head using scanning laser tomography. No abnormalities in Blue-Yellow or Red-Green colour CS was apparent for the OHT patients. A greater vulnerability of the SWS pathway in glaucoma was demonstrated using Blue-Yellow perimetry however predicting which patients may benefit from B-Y perimetric examination is difficult. Furthermore, cataract and the extent of the field loss may limit the extent to which the integrity of the SWS channels can be selectively examined.

Non-standard techniques for the investigation of the visual field

The study investigated the potential applications and the limitations of non-standard techniques of visual field investigation utilizing automated perimetry. Normal subjects exhibited a greater sensitivity to kinetic stimuli than to static stimuli of identical size. The magnitude of physiological SKD was found to be largely independent of age, stimulus size, meridian and eccentricity. The absence of a dependency on stimulus size indicated that successive lateral spatial summation could not totally account for the underlying mechanism of physiological SKD. The visual field indices MD and LV exhibited a progressive deterioration during the time course of a conventional central visual field examination both for normal subjects and for ocular hypertensive patients. The fatigue effect was more pronounced in the latter stages and for the second eye tested. The confidence limits for the definition of abnormality should reflect the greater effect of fatigue on the second eye. A 330 cdm-2 yellow background was employed for blue-on-yellow perimetry. Instrument measurement range was preserved by positioning a concave mirror behind the stimulus bulb to increase the light output by 60% . The mean magnitude of SWS pathway isolation was approximately 1.4 log units relative to a 460nm stimulus filter. The absorption spectra of the ocular media exhibited an exponential increase with increase in age, whilst that of the macular pigment showed no systematic trend. The magnitude of ocular media absorption was demonstrated to reduce with increase in wavelength. Ocular media absorption was significantly greater in diabetic patients than in normal subjects. Five diabetic patients with either normal or borderline achromatic sensitivity exhibited an abnormal blue-on-yellow sensitivity; two of these patients showed no signs of retinopathy. A greater vulnerability of the SWS pathway to the diabetic disease process was hypothesized.

Agreement and clinical comparison between a new swept-source optical coherence tomography-based optical biometer and an optical low-coherence reflectometry biometer

Purpose To compare measurements taken using a swept-source optical coherence tomography-based optical biometer (IOLmaster 700) and an optical low-coherence reflectometry biometer (Lenstar 900), and to determine the clinical impacts of differences in their measurements on intraocular lens (IOL) power predictions. Methods Eighty eyes of 80 patients scheduled to undergo cataract surgery were examined with both biometers. The measurements made using each device were axial length (AL), central corneal thickness (CCT), aqueous depth (AQD), lens thickness (LT), mean keratometry (MK), white-to-white distance (WTW), and pupil diameter (PD). Holladay 2 and SRK/T formulas were used to calculate IOL power. Differences in measurement between the two biometers were determined using the paired t-test. Agreement was assessed through intraclass correlation coefficients (ICC) and Bland–Altman plots. Results Mean patient age was 76.3±6.8 years (range 59–89). Using the Lenstar, AL and PD could not be measured in 12.5 and 5.25% of eyes, respectively, while IOLMaster 700 took all measurements in all eyes. The variables CCT, AQD, LT, and MK varied significantly between the two biometers. According to ICCs, correlation between measurements made with both devices was excellent except for WTW and PD. Using the SRK/T formula, IOL power prediction based on the data from the two devices were statistically different, but differences were not clinically significant. Conclusions No clinically relevant differences were detected between the biometers in terms of their measurements and IOL power predictions. Using the IOLMaster 700, it was easier to obtain biometric measurements in eyes with less transparent ocular media or longer AL.

Modelling ocular monochromatic aberrations using schematic eyes with homogeneous optical media

Previous research has indicated that schematic eyes incorporating aspheric surfaces but lacking gradient index are unable to model ocular spherical aberration and peripheral astigmatism simultaneously. This limits their use as wide-angle schematic eyes. This thesis challenges this assumption by investigating the flexibility of schematic eyes comprising aspheric optical surfaces and homogeneous optical media. The full variation of ocular component dimensions found in human eyes was established from the literature. Schematic eye parameter variants were limited to these dimensions. The levels of spherical aberration and peripheral astigmatism modelled by these schematic eyes were compared to the range of measured levels. These were also established from the literature. To simplify comparison of modelled and measured data, single value parameters were introduced; the spherical aberration function (SAF), and peripheral astigmatism function (PAF). Some ocular components variations produced a wide range of aberrations without exceeding the limits of human ocular components. The effect of ocular component variations on coma was also investigated, but no comparison could be made as no empirical data exists. It was demonstrated that by combined manipulation of a number of parameters in the schematic eyes it was possible to model all levels of ocular spherical aberration and peripheral astigmatism. However, the unique parameters of a human eye could not be obtained in this way, as a number of models could be used to produce the same spherical aberration and peripheral astigmatism, while giving very different coma levels. It was concluded that these schematic eyes are flexible enough to model the monochromatic aberrations tested, the absence of gradient index being compensated for by altering the asphericity of one or more surfaces.