Single Retinal Cells Research Articles

Real-Time Imaging of Single Retinal Cell Apoptosis in a Non-Human Primate Ocular Hypertension Model.

To evaluate the feasibility of using DARC (detection of apoptosing retinal cells) technology as a biomarker for preclinical assessment of glaucomatous damage in a non-human primate (NHP) model of ocular hypertension (OHT). Elevated intraocular pressure (IOP) was induced by applying a laser to the trabecular meshwork in each eye of NHPs. Changes in DARC counts in the retina, identified as fluorescent-tagged annexin V (ANX776)-positive cells, were evaluated together with optic nerve damage, assessed using spectral domain-optical coherence tomography. The pharmacokinetic properties of ANX776 in both healthy and OHT model monkeys were also examined. Sustained elevation of IOP and subsequent thinning of the retinal nerve fiber layer thickness (RNFLT) around the optic nerve head were confirmed in the OHT model. Increases in DARC counts were also detected after IOP elevation. We identified a statistically significant relationship between cumulative DARC counts and reductions in RNFLT both globally and in each peripapillary sector. Intravenous administration of ANX776 increased blood annexin V in a dose-dependent manner, which was subsequently eliminated. This study revealed that DARC technology can effectively assess glaucomatous damage in an NHP OHT model. We obtained the fundamental data that could serve as a reference for developing preclinical models to evaluate the pharmacodynamics of neuroprotective agents using DARC technology in NHP OHT models. Our basic data in a monkey OHT model could be useful for future preclinical studies using DARC technology to estimate the pharmacodynamic response of neuroprotective agents.

Detecting retinal neural and stromal cell classes and ganglion cell subtypes based on transcriptome data with deep transfer learning.

To develop and assess the accuracy of deep learning models that identify different retinal cell types, as well as different retinal ganglion cell (RGC) subtypes, based on patterns of single-cell RNA sequencing (scRNA-seq) in multiple datasets. Deep domain adaptation models were developed and tested using three different datasets. The first dataset included 44 808 single retinal cells from mice (39 cell types) with 24 658 genes, the second dataset included 6225 single RGCs from mice (41 subtypes) with 13 616 genes and the third dataset included 35 699 single RGCs from mice (45 subtypes) with 18 222 genes. We used four loss functions in the learning process to align the source and target distributions, reduce misclassification errors and maximize robustness. Models were evaluated based on classification accuracy and confusion matrix. The accuracy of the model for correctly classifying 39 different retinal cell types in the first dataset was ∼92%. Accuracy in the second and third datasets reached ∼97% and 97% in correctly classifying 40 and 45 different RGCs subtypes, respectively. Across a range of seven different batches in the first dataset, the accuracy of the lead model ranged from 74% to nearly 100%. The lead model provided high accuracy in identifying retinal cell types and RGC subtypes based on scRNA-seq data. The performance was reasonable based on data from different batches as well. The validated model could be readily applied to scRNA-seq data to identify different retinal cell types and subtypes. The code and datasets are available on https://github.com/DM2LL/Detecting-Retinal-Cell-Classes-and-Ganglion-Cell-Subtypes. We have also added the class labels of all samples to the datasets. Supplementary data are available at Bioinformatics online.

Revealing Fine Structures of the Retinal Receptive Field by Deep-Learning Networks.

Deep convolutional neural networks (CNNs) have demonstrated impressive performance on many visual tasks. Recently, they became useful models for the visual system in neuroscience. However, it is still not clear what is learned by CNNs in terms of neuronal circuits. When a deep CNN with many layers is used for the visual system, it is not easy to compare the structure components of CNNs with possible neuroscience underpinnings due to highly complex circuits from the retina to the higher visual cortex. Here, we address this issue by focusing on single retinal ganglion cells with biophysical models and recording data from animals. By training CNNs with white noise images to predict neuronal responses, we found that fine structures of the retinal receptive field can be revealed. Specifically, convolutional filters learned are resembling biological components of the retinal circuit. This suggests that a CNN learning from one single retinal cell reveals a minimal neural network carried out in this cell. Furthermore, when CNNs learned from different cells are transferred between cells, there is a diversity of transfer learning performance, which indicates that CNNs are cell specific. Moreover, when CNNs are transferred between different types of input images, here white noise versus natural images, transfer learning shows a good performance, which implies that CNNs indeed capture the full computational ability of a single retinal cell for different inputs. Taken together, these results suggest that CNNs could be used to reveal structure components of neuronal circuits, and provide a powerful model for neural system identification.

Single-cell transcriptional logic of cell-fate specification and axon guidance in early-born retinal neurons.

Retinal ganglion cells (RGCs), cone photoreceptors (cones), horizontal cells and amacrine cells are the first classes of neurons produced in the retina. However, an important question is how this diversity of cell states is transcriptionally produced. Here, we profiled 6067 single retinal cells to provide a comprehensive transcriptomic atlas showing the diversity of the early developing mouse retina. RNA velocities unveiled the dynamics of cell cycle coordination of early retinogenesis and define the transcriptional sequences at work during the hierarchical production of early cell-fate specification. We show that RGC maturation follows six waves of gene expression, with older-generated RGCs transcribing increasing amounts of guidance cues for young peripheral RGC axons that express the matching receptors. Spatial transcriptionally deduced features in subpopulations of RGCs allowed us to define novel molecular markers that are spatially restricted. Finally, the isolation of such a spatially restricted population, ipsilateral RGCs, allowed us to identify their molecular identity at the time they execute axon guidance decisions. Together, these data represent a valuable resource shedding light on transcription factor sequences and guidance cue dynamics during mouse retinal development.

A pilot study assessing retinal pathology in psychosis using optical coherence tomography: Choroidal and macular thickness

Mounting evidence supports a genetic-vascular-inflammatory etiology of schizophrenia. The retina provides an indirect assessment of inflammation and degeneration in the brain. In particular, the use of spectral domain optical coherence tomography (SD-OCT) has emerged as a powerful tool for examining single retinal nerve cell layers and the choroid, the vascular layer supplying the outer retina. In this study, choroidal and macular thicknesses were measured in six patients with psychosis with either schizophrenia or bipolar disorder, and in 18 age- and sex-matched healthy controls. Mean choroidal thickness was reduced in psychosis, though not significantly so. There was a statistically significant decrease in macular thickness in psychosis patients predominantly affecting the inner layers of the macula. Significant macular thinning may signal vascular, inflammatory, or degenerative processes that may also be occurring in the brain. This is one of the first studies to examine choroidal thickness in psychosis. Further studies are needed to determine whether the retinal changes in psychosis are correlated with microvascular dysfunction, neuroinflammation, and neurodegeneration.

AP-2\u03b5 Expression in Developing Retina: Contributing to the Molecular Diversity of Amacrine Cells

AP-2 transcription factors play important roles in the regulation of gene expression during development. Four of the five members of the AP-2 family (AP-2α, AP-2β, AP-2γ and AP-2δ) have previously been shown to be expressed in developing retina. Mouse knockouts have revealed roles for AP-2α, AP-2β and AP-2δ in retinal cell specification and function. Here, we show that the fifth member of the AP-2 family, AP-2ε, is also expressed in amacrine cells in developing mammalian and chicken retina. Our data indicate that there are considerably fewer AP-2ε-positive cells in the developing mouse retina compared to AP-2α, AP-2β and AP-2γ-positive cells, suggesting a specialized role for AP-2ε in a subset of amacrine cells. AP-2ε, which is restricted to the GABAergic amacrine lineage, is most commonly co-expressed with AP-2α and AP-2β, especially at early stages of retinal development. Co-expression of AP-2ε and AP-2γ increases with differentiation. Analysis of previously published Drop-seq data from single retinal cells supports co-expression of multiple AP-2s in the same cell. Since AP-2s bind to their target sequences as either homodimers or heterodimers, our work suggests spatially- and temporally-coordinated roles for combinations of AP-2 transcription factors in amacrine cells during retinal development.

Development of Refractive Errors—What Can We Learn From Inherited Retinal Dystrophies?

It is unknown which retinal cells are involved in the retina-to-sclera signaling cascade causing myopia. As inherited retinal dystrophies (IRD) are characterized by dysfunction of a single retinal cell type and have a high risk of refractive errors, a study investigating the affected cell type, causal gene, and refractive error in IRDs may provide insight herein. Case-control study. Study Population: Total of 302 patients with IRD from 2 ophthalmogenetic centers in the Netherlands. Reference Population: Population-based Rotterdam Study-III and Erasmus Rucphen Family Study (N= 5550). Distributions and mean spherical equivalent (SE) were calculated for main affected cell type and causal gene; and risks of myopia and hyperopia were evaluated using logistic regression. Bipolar cell-related dystrophies were associated with the highest risk of SE high myopia 239.7; odds ratio (OR) mild hyperopia 263.2, both P < .0001; SE-6.86 diopters (D) (standard deviation [SD] 6.38), followed by cone-dominated dystrophies (OR high myopia 19.5, P < .0001; OR high hyperopia 10.7, P= .033; SE-3.10 D [SD 4.49]); rod dominated dystrophies (OR high myopia 10.1, P < .0001; OR high hyperopia 9.7, P= .001; SE-2.27 D [SD 4.65]), and retinal pigment epithelium (RPE)-related dystrophies (OR low myopia 2.7; P= .001; OR high hyperopia 5.8; P= .025; SE-0.10 D [SD 3.09]). Mutations in RPGR (SE-7.63 D [SD 3.31]) and CACNA1F (SE-5.33 D [SD 3.10]) coincided with the highest degree of myopia and in CABP4 (SE 4.81 D [SD 0.35]) with the highest degree of hyperopia. Refractive errors, in particular myopia, are common in IRD. The bipolar synapse and the inner and outer segments of the photoreceptor may serve as critical sites for myopia development.

GABAB receptors enhance excitatory responses in isolated rat retinal ganglion cells.

GABA is an inhibitory transmitter but can sometimes produce paradoxical excitatory effects through synaptic networks. We found a novel GABA-mediated excitation within a single retinal cell. It involves a chain of events from receptor stimulation to the sequential modulation of two associated channels, resulting in enhanced neuroexcitability. GABAB receptor activation selectively suppresses N-type calcium channels. The BK-type potassium channels are exclusively linked to the N-type calcium channel. Thus, stimulation of GABAB receptors suppresses an outward current, increasing the excitatory range of single neurons. GABAB receptors (GABAB Rs) suppress voltage-gated calcium channels and activate G-protein coupled potassium channels (GIRK and TREK channels), both mechanisms serving to inhibit neurons. In isolated rat retinal spiking neurons, GABAB Rs produce both actions but the net effect is to enhance excitatory signals. This is because GABAB Rs selectively suppress N-type calcium channels, which in turn are specifically linked to BK channels. Consequently, when GABAB Rs are stimulated there is a reduction in outward current, allowing neurons to extend their level of depolarization. Whereas many retinal neurons use L-type channels to stimulate vesicle fusion, the suppression of N-type channels augments dynamic range without affecting transmitter release.

Retinal Wave Patterns Are Governed by Mutual Excitation among Starburst Amacrine Cells and Drive the Refinement and Maintenance of Visual Circuits.

Spontaneous activity drives early mammalian circuit development, but the initiation and patterning of activity vary across development and among modalities. Cholinergic "retinal waves" are initiated in starburst amacrine cells and propagate to retinal ganglion cells and higher-order visual areas, but the mechanism responsible for creating their unique and critical activity pattern is incompletely understood. We demonstrate that cholinergic wave patterns are dictated by recurrent connectivity within starburst amacrine cells, and retinal ganglion cells act as "readouts" of patterned activity. We also show that eye-specific segregation occurs normally without glutamatergic waves, but elimination of both cholinergic and glutamatergic waves completely disrupts visual circuit development. These results suggest that each retinal wave pattern during development is optimized for concurrently refining multiple visual circuits.

Analysis of gene expression in wild-type and Notch1 mutant retinal cells by single cell profiling.

The vertebrate retina comprises sensory neurons, the photoreceptors, as well as many other types of neurons and one type of glial cell. These cells are generated by multipotent and restricted retinal progenitor cells (RPCs), which express Notch1. Loss of Notch1 in RPCs late during retinal development results in the overproduction of rod photoreceptors at the expense of interneurons and glia. To examine the molecular underpinnings of this observation, microarray analysis of single retinal cells from wild-type or Notch1 conditional knockout retinas was performed. In situ hybridization was carried out to validate some of the findings. The majority of Notch1-mutant cells lost expression of known Notch target genes. These cells also had low levels of RPC and cell cycle genes, and robustly up-regulated rod precursor genes. In addition, single wild-type cells, in which cell cycle marker genes were down-regulated, expressed markers of both rod photoreceptors and interneurons.

Notch1 is required in newly postmitotic cells to inhibit the rod photoreceptor fate

Several models of cell fate determination can be invoked to explain how single retinal progenitor cells (RPCs) produce different cell types in a terminal division. To gain insight into this process, the effects of the removal of a cell fate regulator, Notch1, were studied in newly postmitotic cells using a conditional allele of Notch1 (N1-CKO) in mice. Almost all newly postmitotic N1-CKO cells became rod photoreceptors, whereas wild-type (WT) cells achieved a variety of fates. Single cell profiling of wild-type and N1-CKO retinal cells transitioning from progenitor to differentiated states revealed differential expression of inhibitor of DNA binding factors Id1 and Id3, as well as Notch-regulated ankyrin repeat protein (Nrarp). Misexpression of Id1 and Id3 was found to be sufficient to drive production of Müller glial cells and/or RPCs. Moreover, Id1 and Id3 were shown to partially rescue the production of bipolar and Müller glial cells in the absence of Notch1 in mitotic and newly postmitotic cells. Misexpression of Nrarp, a downstream target gene and inhibitor of the Notch signaling pathway, resulted in the overproduction of rod photoreceptors at the expense of Müller glial cells. These data demonstrate that cell fate decisions can be made in newly postmitotic retinal cells, and reveal some of the regulators downstream of Notch1 that influence the choice of rod and non-rod fates. Taken together, our results begin to address how different signals downstream from a common pathway lead to different fate outcomes.

Clinical Applications of High Resolution In-Vivo Retinal Imaging

Clinical Applications of High Resolution In-Vivo Retinal Imaging

Single-cell Profiling of Developing and Mature Retinal Neurons

Highly specialized, but exceedingly small populations of cells play important roles in many tissues. The identification of cell-type specific markers and gene expression programs for extremely rare cell subsets has been a challenge using standard whole-tissue approaches. Gene expression profiling of individual cells allows for unprecedented access to cell types that comprise only a small percentage of the total tissue(1-7). In addition, this technique can be used to examine the gene expression programs that are transiently expressed in small numbers of cells during dynamic developmental transitions(8). This issue of cellular diversity arises repeatedly in the central nervous system (CNS) where neuronal connections can occur between quite diverse cells(9). The exact number of distinct cell types is not precisely known, but it has been estimated that there may be as many as 1000 different types in the cortex itself(10). The function(s) of complex neural circuits may rely on some of the rare neuronal types and the genes they express. By identifying new markers and helping to molecularly classify different neurons, the single-cell approach is particularly useful in the analysis of cell types in the nervous system. It may also help to elucidate mechanisms of neural development by identifying differentially expressed genes and gene pathways during early stages of neuronal progenitor development. As a simple, easily accessed tissue with considerable neuronal diversity, the vertebrate retina is an excellent model system for studying the processes of cellular development, neuronal differentiation and neuronal diversification. However, as in other parts of the CNS, this cellular diversity can present a problem for determining the genetic pathways that drive retinal progenitors to adopt a specific cell fate, especially given that rod photoreceptors make up the majority of the total retinal cell population(11). Here we report a method for the identification of the transcripts expressed in single retinal cells (Figure 1). The single-cell profiling technique allows for the assessment of the amount of heterogeneity present within different cellular populations of the retina(2,4,5,12). In addition, this method has revealed a host of new candidate genes that may play role(s) in the cell fate decision-making processes that occur in subsets of retinal progenitor cells(8). With some simple adjustments to the protocol, this technique can be utilized for many different tissues and cell types.

Single-cell Profiling of Developing and Mature Retinal Neurons

Highly specialized, but exceedingly small populations of cells play important roles in many tissues. The identification of cell-type specific markers and gene expression programs for extremely rare cell subsets has been a challenge using standard whole-tissue approaches. Gene expression profiling of individual cells allows for unprecedented access to cell types that comprise only a small percentage of the total tissue1-7. In addition, this technique can be used to examine the gene expression programs that are transiently expressed in small numbers of cells during dynamic developmental transitions8. This issue of cellular diversity arises repeatedly in the central nervous system (CNS) where neuronal connections can occur between quite diverse cells9. The exact number of distinct cell types is not precisely known, but it has been estimated that there may be as many as 1000 different types in the cortex itself10. The function(s) of complex neural circuits may rely on some of the rare neuronal types and the genes they express. By identifying new markers and helping to molecularly classify different neurons, the single-cell approach is particularly useful in the analysis of cell types in the nervous system. It may also help to elucidate mechanisms of neural development by identifying differentially expressed genes and gene pathways during early stages of neuronal progenitor development. As a simple, easily accessed tissue with considerable neuronal diversity, the vertebrate retina is an excellent model system for studying the processes of cellular development, neuronal differentiation and neuronal diversification. However, as in other parts of the CNS, this cellular diversity can present a problem for determining the genetic pathways that drive retinal progenitors to adopt a specific cell fate, especially given that rod photoreceptors make up the majority of the total retinal cell population11. Here we report a method for the identification of the transcripts expressed in single retinal cells (Figure 1). The single-cell profiling technique allows for the assessment of the amount of heterogeneity present within different cellular populations of the retina2,4,5,12. In addition, this method has revealed a host of new candidate genes that may play role(s) in the cell fate decision-making processes that occur in subsets of retinal progenitor cells8. With some simple adjustments to the protocol, this technique can be utilized for many different tissues and cell types.

Fate Restriction and Multipotency in Retinal Stem Cells

Stem cells have the capacity to both self-renew and generate postmitotic cells. Long-term tracking of individual clones in their natural environment constitutes the ultimate way to validate postembryonic stem cells. We identify retinal stem cells (RSCs) using the spatiotemporal organization of the fish retina and follow the complete offspring of a single cell during the postnatal life. RSCs generate two tissues of the adult fish retina, the neural retina (NR) and the retinal-pigmented epithelium (RPE). Despite their common embryonic origin and tight coordination during continuous organ growth, we prove that NR and RPE are maintained by dedicated RSCs that contribute in a fate-restricted manner to either one or the other tissue. We show that in the NR, RSCs are multipotent and generate all neuron types and glia. The clonal origin of these different cell types from a multipotent NSC has far-reaching implications for cell type and tissue homeostasis.

Imaging apoptosis in the eye

Apoptosis is a form of programmed cell death that is implicated in both pathological and physiological processes throughout the body. Its imaging in vivo with intravenous radiolabelled-annexin V has been heralded as an important advance, with around 30 clinical trials demonstrating its application in the early detection and monitoring of disease, and the assessment of efficacy of potential and existing therapies. A recent development has been the use of fluorescently labeled annexin V to visualize single retinal cells undergoing the process of apoptosis in vivo with ophthalmoscopy. This has been given the acronym DARC (Detection of Apoptosing Retinal Cells). DARC so far has only been used experimentally, but clinical trials are starting shortly in glaucoma patients. Results suggest that DARC may provide a direct assessment of retinal ganglion cell health. By enabling early assessment and quantitative analysis of cellular degeneration in glaucoma, it is hoped that DARC can identify patients before the onset of irreversible vision loss. Furthermore, in addition to aiding the tracking of disease, it may provide a rapid and objective assessment of potential and effective therapies, providing a new and meaningful clinical endpoint in glaucomatous disease that is so badly needed.

In vivo imaging of microscopic structures in the rat retina.

The ability to resolve single retinal cells in rodents in vivo has applications in rodent models of the visual system and retinal disease. The authors have characterized the performance of a fluorescence adaptive optics scanning laser ophthalmoscope (fAOSLO) that provides cellular and subcellular imaging of rat retina in vivo. Enhanced green fluorescent protein (eGFP) was expressed in retinal ganglion cells of normal Sprague-Dawley rats via intravitreal injections of adeno-associated viral vectors. Simultaneous reflectance and fluorescence retinal images were acquired using the fAOSLO. fAOSLO resolution was characterized by comparing in vivo images with subsequent imaging of retinal sections from the same eyes using confocal microscopy. Retinal capillaries and eGFP-labeled ganglion cell bodies, dendrites, and axons were clearly resolved in vivo with adaptive optics. Adaptive optics correction reduced the total root mean square wavefront error, on average, from 0.30 microm to 0.05 microm (measured at 904 nm, 1.7-mm pupil). The full width at half maximum (FWHM) of the average in vivo line-spread function (LSF) was approximately 1.84 microm, approximately 82% greater than the FWHM of the diffraction-limited LSF. With perfect aberration compensation, the in vivo resolution in the rat eye could be approximately 2x greater than that in the human eye because of its large numerical aperture (approximately 0.43). Although the fAOSLO corrects a substantial fraction of the rat eye's aberrations, direct measurements of retinal image quality reveal some blur beyond that expected from diffraction. Nonetheless, subcellular features can be resolved, offering promise for using adaptive optics to investigate the rodent eye in vivo with high resolution.

Proliferation and Expression of Progenitor and Mature Retinal Phenotypes in the Adult Mammalian Ciliary Body after Retinal Ganglion Cell Injury

Despite the identification of a small population of cells residing in the ciliary body (CB) of the adult mammalian eye that have the capacity to generate retina-like cells in vitro, their activity in vivo remains quiescent. The authors sought to identify whether the predictable and time-dependent death of retinal ganglion cells (RGCs) results in activation of progenitor-like cells within the CB. RGC injury was induced by optic nerve axotomy in adult mice. Thymidine-analogue lineage tracing and immunocytochemistry were used to identify dividing cells and the phenotype of newly generated progeny. Two populations of nestin-expressing cells are present in the CB of the uninjured eye. One population resides in periendothelial cells of blood vessels, and a second resides in the ciliary epithelium. Axotomy increases proliferation in the CB, a response that begins before the onset of RGC death and continues during a time that corresponds with the peak in RGC death. In addition, a subpopulation of nestin-positive cells in the CB upregulates the homeodomain protein Chx10. Finally, recoverin, the expression of which is normally restricted to photoreceptors and bipolar cells of the retina, is upregulated in the CB in a manner that is independent of proliferation. Together, these results suggest that progenitorlike cells of the CB respond to cues associated with the loss of a single retinal cell type and that a subpopulation of those cells may differentiate into a cell that bears phenotypic resemblance to those seen in the retina.

How to watch ganglion cells in the DARC?

Abstract Purpose: Assessment of neuroprotection in vivo is difficult. DARC (Detection of Apoptosing Retinal Cells) is a newly devised, non‐invasive real‐time imaging technique using confocal laser scanning ophthalmoscopy to visualise single retinal nerve cell apoptosis in vivo. Methods: DARC allows longitudinal study of disease processes, which has not previously been possible. The implications of this novel technique include its application to glaucoma as a powerful new clinical tool with which to diagnose and identify patients with early disease, before they lose vision. Furthermore it may serve as a surrogate biomarker, providing real‐time information that could dramatically reduce the duration of glaucoma clinical studies, which currently have to use visual field status as a key endpoint and determinant of outcome. Results: We have been able for the first time to image changes occurring in neuronal cell apoptosis over hours, days and months, and show effects depending on the magnitude of the initial apoptotic inducer in several experimental models of neurodegeneration. Conclusions: More importantly, by using the eye as a “window” onto the brain, DARC opens the door to directly observing effects of therapeutic strategies in neurodegenerative disorders, by allowing for the study of potential neuroprotective drugs using meaningful endpoints that are based on the direct assessment of neuronal death in vivo. We have already assessed several neuroprotective agents using DARC including neuropeptide approaches.

Applications of adaptive‐optics in ophthalmology

Abstract Purpose: This presentation reviews the current and potential applications of adaptive optics (AO) in the area of ophthalmology. Methods: AO was invented by astrophysicists in order to eliminate image blur and twinkling in large terrestrial telescopes. This technique cancels the optical distortions that are introduced by the earth atmosphere, a turbid and inhomogeneous medium, in the light emitted by distant stars. Like the atmosphere, the human eye is optically imperfect: it suffers from wavefront aberrations whose pattern and amount differ between individuals. This is the reason why AO has met several applications in ophthalmology. One part of the AO technology consists in measuring the distortions of light using a wavefront sensor: this technique has been integrated into wavefront aberrometers that routinely improve the precision of LASIK procedures. Results: The full implementation of AO has recently led to other breakthrough devices. AO visual simulators are enhanced aberrometers that allow patients to visualize, at the pre‐operative stage, the expected outcome of refractive surgery procedures, including presbyopic ablations. Several research laboratories have implemented AO to improve the resolution of various retinal imaging techniques (fundus camera, SLO, OCT) up to the scale of single retinal cells. Three‐dimensional images of cone photoreceptors have recently been obtained with a resolution of 3μm, both in the longitudinal and transverse dimensions of the retinal tissue. Aside from this world‐record, the presentation will also address the limitations of the technique and describe some of the work in progress towards the clinical applications of AO technology. Conclusions: AO technology will likely provide new devices that meet unsatisfied needs in refractive surgery and retinal disagnostics.