In vitro ophthalmic ultrahigh-resolution OCT imaging reveals retinal morphology with unprecedented detail. The specific assignment of OCT signal patterns to retinal substructures provides a basis for improved interpretation of in vivo ophthalmic OCT tomograms of high clinical relevance.
Noncontact, depth-resolved, optical probing of retinal response to visual stimulation with a <10-m spatial resolution, achieved by using functional ultrahigh-resolution optical coherence tomography (fUHROCT), is demonstrated in isolated rabbit retinas. The method takes advantage of the fact that physiological changes in dark-adapted retinas caused by light stimulation can result in local variation of the tissue reflectivity. fUHROCT scans were acquired from isolated retinas synchronously with electrical recordings before, during, and after light stimulation. Pronounced stimulusrelated changes in the retinal reflectivity profile were observed in the inner͞outer segments of the photoreceptor layer and the plexiform layers. Control experiments (e.g., dark adaptation vs. light stimulation), pharmacological inhibition of photoreceptor function, and synaptic transmission to the inner retina confirmed that the origin of the observed optical changes is the altered physiological state of the retina evoked by the light stimulus. We have demonstrated that fUHROCT allows for simultaneous, noninvasive probing of both retinal morphology and function, which could significantly improve the early diagnosis of various ophthalmic pathologies and could lead to better understanding of pathogenesis.electroretinogram ͉ functional optical coherence tomography ͉ inner plexiform layer ͉ photoreceptors ͉ retinal imaging T he vertebrate retina consists of several distinct layers: nuclear layers containing cell bodies can be differentiated from plexiform layers with axons and dendrites forming the neuronal network that preprocesses light-evoked signals before transmission to the brain. Early stages of retinal disorders are often confined to one of these layers and are manifested by both morphological abnormalities and impaired physiological responses. Detection of such pathologies requires high-resolution imaging methods. Various imaging modalities such as fundus photography, ultrasound imaging, and optical coherence tomography (OCT) are clinically used for imaging retinal morphology. OCT is an emerging imaging technique that allows for noncontact, in vivo visualization of biological tissue morphology with a micrometer-scale resolution at imaging depths of 1-2 mm (1-3). Currently, electrophysiological tests such as electroretinography (ERG) (4) and multifocal ERG (5) are used for clinical assessment of retinal function.More then 25 years ago, it was observed that the isolated retina when stimulated with visible light changes the amount of transmitted near-infrared light (NIR) (6, 7). Photoreceptors (PRs) were determined to be the main source of this effect, and in the following years, this method was used for investigation and quantitative evaluation of the activation of the PR G protein transducin and the time course of transduction events (8-10 and reviewed in ref. 11). In the last few years, other physiological processes at the cellular and subcellular level such as membrane depolarization (12), cell swelling (13), and altered metabolism...
A minor population of cone photoreceptors (called B-cones) can be distinguished from the major population (called R-cones) on morphological criteria as seen by light microscopy in foveal and peripheral human retina. The B-cones are characterized by a longer inner segment projecting into subretinal space, a larger-diameter inner segment, an increased staining intensity of the inner segment, and a different distribution relative to the R-cones in the cone mosaic. B-cones occur even in the foveolar center (3-5%) and rise to a maximum (15%) in the foveolar slope. They can also be identified in peripheral retina where they form 7-10% of the total cone population. The B-cone population follows the distribution profile postulated for the blue-sensitive system from histochemical studies on monkeys and from psychophysical studies on humans. The B-cones also share many of the same morphological features of the putative blue cones of the ground squirrel and monkey retinas. For these reasons we suggest that our B-cone group is the blue cone population of the human retina.
This study addresses the correlation of retinal topography with factors such as the visual environment, life style, and behavior for a major mammalian group, the artiodactyls. To provide a broader basis for semiquantitative comparison, short-wavelength-sensitive (S)- and middle-to-long-wavelength-sensitive (M)-opsin cone receptor populations from 25 species from five artiodactyl families and of the African elephant were labeled and sampled. The resulting topographic maps were analyzed with respect to the position and extension of high-density regions. For better parameter differentiation, systematic relationships were statistically normalized. In all species examined, two classes of cones have been detected. In most species, the S-cone maxima were located in the temporodorsal retina, but there are exceptions such as the roe deer with accumulation in the ventral retina. For M-cones, as a consequence of their role in terrain/food assessment and predator detection, the standard topography is L-shaped: a horizontal visual streak including a temporal area centralis is extended by a temporal rim. Its extension is correlated with the animal's body height (P = 0.0017): small species (pudu, mouse deer) tend to have a visual streak only, whereas the giraffe shows a complete dorsal arch of elevated densities. Furthermore, a size-independent habitat correlation was revealed for a similar M-cone pattern (P < 0.0001): mountainous species show a striking specialization around the dorsal retina, pointing to the importance of the inferior visual field in precipitous terrain.
Cellular in vivo visualization of the three dimensional architecture of individual human foveal cone photoreceptors is demonstrated by combining ultrahigh resolution optical coherence tomography and a novel adaptive optics modality. Isotropic resolution in the order of 2-3 microm, estimated from comparison with histology, is accomplished by employing an ultrabroad bandwidth Titanium:sapphire laser with 140 nm bandwidth and previous correction of chromatic and monochromatic ocular aberrations. The latter, referred to as pancorrection, is enabled by the simultaneous use of a specially designed lens and an electromagnetically driven deformable mirror with unprecedented stroke for correcting chromatic and monochromatic aberrations, respectively. The increase in imaging resolution allows for resolving structural details of distal elements of individual foveal cones: inner segment zones--myoids and ellipsoids--are differentiated from outer segments protruding into pigment epithelial processes in the retina. The presented technique has the potential to unveil photoreceptor development and pathogenesis as well as improved therapy monitoring of numerous retinal diseases.
This paper presents a successful combination of ultra-high speed (120,000 depth scans/s), ultra-high resolution optical coherence tomography with adaptive optics and an achromatizing lens for compensation of monochromatic and longitudinal chromatic ocular aberrations, respectively, allowing for non-invasive volumetric imaging in normal and pathologic human retinas at cellular resolution. The capability of this imaging system is demonstrated here through preliminary studies by probing cellular intraretinal structures that have not been accessible so far with in vivo, non-invasive, label-free imaging techniques, including pigment epithelial cells, micro-vasculature of the choriocapillaris, single nerve fibre bundles and collagenous plates of the lamina cribrosa in the optic nerve head. In addition, the volumetric extent of cone loss in two colour-blinds could be quantified for the first time. This novel technique provides opportunities to enhance the understanding of retinal pathogenesis and early diagnosis of retinal diseases.
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