Photoreceptors in the retina are specialized neuronal cells that perceive light and play a central role in the visual system. Damage to photoreceptors is a clinical feature often associated with various retinal degenerative disorders. The photoreceptor bed comprises a unique extracellular matrix (ECM) scaffold often described as the interphotoreceptor matrix (IPM) in the subretinal space, vital during retinal development and homeostasis. In this study, we used focused ion beam scanning electron microscopy (FIB-SEM) and transmission electron microscopy (TEM) to analyze the ultrastructural architecture of the retinal pigmented epithelium (RPE)-photoreceptor complex in mice. Additionally, we describe methods for retinal preparation in EM imaging. TEM images display ultrastructural retina layers, including Bruch\'s membrane and the interdigitation zone (IZ). The 3-dimensional reconstruction of the outer retina revealed individual photoreceptors, the connection between their inner and outer segment via the photoreceptor cilia, and photoreceptor interaction with the RPE ciliary processes. Our findings highlight the importance of FIB-SEM in deciphering the ultrastructural details of RPE-photoreceptor interactions in the IPM complex which are essential for the maintenance of retinal architecture.

UNASSIGNED: Photoreceptor (PR) outer segments, retinal pigment epithelium apical processes, and inter-PR matrix contribute to the interdigitation zone (IZ) of optical coherence tomography (OCT). We hypothesize that this interface degrades over adulthood, in concert with a delay of rod mediated dark adaptation (RMDA). To explore this idea, we determined IZ discernibility and RMDA in younger and older adults.
UNASSIGNED: For this cross-sectional study, eyes of 20 young (20-30 years) and 40 older (≥60 years) participants with normal maculas according to the AREDS 9-step grading system underwent OCT imaging and RMDA testing at 5° superior to the fovea. Custom FIJI plugins enabled analysis for IZ discernibility at 9 eccentricities in 0.5 mm steps on one single horizontal B-scan through the fovea. Locations with discernible IZ met two criteria: visibility on B-scans and a distinct peak on a longitudinal reflectivity profile. The frequency of sites meeting both criteria was compared between both age groups and correlated with rod intercept time (RIT).
UNASSIGNED: The median number of locations with discernible IZ was significantly higher (foveal, 4 vs. 0, p = 0.0099; extra-foveal 6 vs. 0, p < 0.001) in eyes of young (26 ± 3 years) compared to older (73 ± 5 years) participants. For the combined young and older sample, the higher frequency of discernible IZ was correlated with shorter RIT (faster dark adaptation) (rs = -0.56, p < 0.0001). This association was significant within young eyes (rs = -0.54; p = 0.0134) and not within older eyes (rs = -0.29, p = 0.706).
UNASSIGNED: Results suggest that the interface between outer segments and apical processes degrades in normal aging, potentially contributing to delayed rod-mediated dark adaptation. More research is needed to verify an age-related association between IZ discernibility and rod-mediated dark adaptation. If confirmed in a large sample, IZ discernibility might prove to be a valuable biomarker and predictor for visual function in aging.

BACKGROUND: The study aims to investigate the relationship between the volume-accumulated reflectivity (termed \"integral\") on spectral domain optical coherence tomography (SD-OCT) and cone density on adaptive optics (AO) imaging.
METHODS: In this cross-sectional study, both eyes of 32 healthy subjects and 5 patients with inherited retinal diseases (IRD) were studied. The parameter, integral, was defined as the volume-accumulated reflectivity values in a selected region on OCT images; integrals of the ellipsoid zone (EZ) and interdigitation zone (IZ) were measured at 2°, 3°, 4°, 5°and 6° eccentricity along the four meridians on fovea-centered OCT B-scans. Cone density in the same region was measured using a flood illumination adaptive optics camera RTX1.
RESULTS: Integrals of EZ, IZ and cone density shared similar distribution patterns. Integral of the IZ was better correlated with cone density in both healthy people (r = 0.968, p < 0.001) and those with IRD (r = 0.823, p < 0.001) than direct measurements of reflectivity on OCT images. A strong correlation was found between best corrected visual acuity (BCVA) and cone density at 2° eccentricity (r = -0.857, p = 0.002). BCVA was also correlated with the integral of the IZ at the foveola (r = -0.746, p = 0.013) and fovea (r = -0.822, p = 0.004).
CONCLUSIONS: The new parameter \"integral\" of the photoreceptor outer segment measured from SD-OCT was noted to correlate with cone density and visual function in this pilot study.

OBJECTIVE: To investigate the changes in the external limiting membrane (ELM), ellipsoid zone (EZ), and interdigitation zone (IZ) integrity and their relationship with visual outcomes after idiopathic epiretinal membranes peeling.
METHODS: Clinical records of 150 eyes from 144 consecutive patients who underwent vitrectomy were reviewed. The status of IZ, EZ, and ELM was assessed by spectral-domain optical coherence tomography at baseline and 1, 4, 10, and 24 months postoperatively.
RESULTS: Sixty-one eyes presented with photoreceptor layer disruption preoperatively, and IZ disruption (40.7%) was the primary type. The best-corrected visual acuity (BCVA) in the photoreceptor disruption group was significantly lower than that in the intact group at baseline and the final follow-up. Of them, ELM + EZ + IZ disruption showed the worst BCVA (P = 0.001). After surgery, 62 eyes were observed with disruption aggravated. EZ + IZ disruption (51.0%) was the most frequent type at 1 month postoperatively. The eyes with longer symptom duration, better BCVA, earlier stage, thinner CFT at baseline, and combined cataract surgery more tended to be observed with photoreceptor damage progressed after surgery (P < 0.05). There was no significant difference in the final BCVA between the eyes with and without damage progressed (P = 0.332). Finally, 28.1% of the eyes recovered photoreceptor continuity. The eyes with foveal photoreceptor integrity restored had better BCVA than those remaining discontinuous (P < 0.001).
CONCLUSIONS: ERM-induced photoreceptor disruption mainly manifests as IZ disruption and has a negative effect on BCVA, whereas surgery mainly causes EZ and IZ disruption, which does not have a significant impact on the final BCVA.

UNASSIGNED: To determine the changes in the microstructures of the photoreceptors in patients with autosomal recessive bestrophinopathy (ARB) by ultrahigh-resolution spectral-domain optical coherence tomography (UHR-SD-OCT).
UNASSIGNED: Five eyes of 4 patients with ARB were studied. Cross-sectional images of the fovea were recorded by the UHR-SD-OCT system with a depth resolution of <2.0 μm.
UNASSIGNED: The UHR-SD-OCT images revealed changes in the outer retinal structures that were dependent on the severity of the photoreceptor atrophy. There was an increase in the reflectivity and appearance of small hyperreflective dots (HRDs) in the outer segments, followed by an irregularity and decrease in the length of the outer segments, then a disruption of the ellipsoid zone (EZ) band, and appearance of large HRDs corresponding to the segmented ellipsoids. Finally, there was a disappearance of the large HRDs followed by a localized thinning of the outer nuclear layer and appearance of hyperreflective foci above the region of the disrupted EZ.
UNASSIGNED: UHR-SD-OCT can record images that show detailed changes of the microstructures of the photoreceptors at different stages of ARB. These observations should help in determining the mechanisms involved in retinal pathology and should provide important information on the effectiveness of treatments.

UNASSIGNED: To analyze the microstructures of the photoreceptor layer in detail in eyes with occult macular dystrophy (OMD, Miyake\'s disease) by ultrahigh-resolution spectral-domain optical coherence tomography (UHR-SD-OCT).
UNASSIGNED: Twenty-eight normal subjects and 5 patients with OMD of different severities were studied. Cross-sectional images through the fovea were recorded with a UHR-SD-OCT system with a depth resolution of <2.0 μm. In patients with OMD, the UHR-SD-OCT images revealed abnormal photoreceptor microstructures which were not detected in the conventional SD-OCT images. The UHR-SD-OCT images showed that the interdigitation zone (IZ) was not present and the outer segments were hyperreflective with hyperreflective dots (HRDs) aligned like string of pearls during the earlier stages. There was a disruption of the ellipsoid zone (EZ) which appeared as clusters of larger HRDs, and these HRDs became less apparent with increasing time. The outer segments became hyporeflective and rod IZ became apparent with longer duration of the disease process.
UNASSIGNED: The UHR-SD-OCT images show detailed characteristics of the photoreceptor microstructures of different severities during the progression of OMD. These detailed observations will help in understanding the mechanisms involved in the retinal pathology and should provide important information for their treatments.

OBJECTIVE: To report an early stage of gyrate atrophy (GA) of the retina and choroid for which spectral-domain optical coherence tomography (SD-OCT) images revealed an obscured interdigitation zone (IZ).
METHODS: A 13-year-old boy was referred to our department due to blurred vision in his left eye. Best corrected visual acuity was 20/20 and 20/25 in the right and left eye, respectively. Fundus examination revealed scalloped atrophic peripheral chorioretinal lesions in both eyes. Concentrations of plasma and urine ornithine were 1192 nmol/mL and 1930 μmol/g·cre, respectively. Consequently, he was diagnosed with GA. Although Goldmann perimetry found no abnormalities, electroretinogram (ERG) revealed loss of the rod responses and significant attenuation of the cone responses in both eyes. Detailed analysis of the posterior part of the fundus using multifocal electroretinogram showed poor responses, specifically in the nasal macular area of the left eye. SD-OCT showed an obscured IZ corresponding to the attenuated cone response determined by ERG. There was preservation of the retinal pigment epithelium, ellipsoid zone and external limiting membrane.
CONCLUSIONS: Obscured IZ during early stage GA was confirmed in a teenage patient with a chief complaint of blurred vision. OCT is useful in the detection of minute morphological changes that occur earlier in GA.

The purpose of this study was to design a new parameter \"integral\" to quantitatively evaluate the spatial cumulative reflectivity of the outer retinal layers in optical coherence tomography (OCT), and to investigate its role in the detection of outer retinal diseases.
This was a cross-sectional study. Fovea-centered line OCT scans were performed on 60 eyes of 60 healthy volunteers and 44 eyes of 44 patients diagnosed with outer retinal diseases. The integrals of the ellipsoid zone (EZ) and interdigitation zone (IZ) were measured by respectively accumulating the grayscale values of all the pixels within the EZ and IZ at specified locations on the scanning lines, and were then adjusted by calculating their percentages on the outer retina. The integrals of the EZ and IZ were compared between the two groups.
The integrals of the EZ and IZ were stably and normally distributed in the healthy eyes, and were significantly lower in eyes with outer retinal lesions than in healthy ones (P < 0.05). Moreover, the integrals of the EZ and IZ were correlated with best corrected visual acuity (BCVA; adjusted R2 = 0.620) and the presence of outer retinal lesions (Nagelkerke R2 = 0.767). The area under the receiver operating characteristic (ROC) curve was 0.954 (95% confidence interval [CI] = 0.918-0.990) when the integral was selected as a diagnostic variable.
Obtained from this novel quantification method, the new parameter integral was comparable between different individuals and had the potential to detect outer retinal abnormalities in reflectivity through OCT.
Our work verified the feasibility of the new image analysis technique in the detection of the diseases affecting the outer retina.

The aim of the current study was to evaluate the correlation between the integrity of the outer retinal layers on optical coherence tomography (OCT) and objective parameters of retinal microvascular perfusion on optical coherence tomography angiography (OCTA).
A total of 105 eyes of 54 diabetic patients were included in the study. Integrity of the outer retinal layers including the external limiting membrane (ELM), ellipsoid zone (EZ), and interdigitation zone (IZ) was assessed by spectral-domain optical coherence tomography. The foveal avascular zone (FAZ) area and vessel density (VD) measurements in the superficial capillary plexus (SCP) and deep capillary plexus (DCP) in all the Early Treatment Diabetic Retinopathy Study (ETDRS) sectors were evaluated by OCTA. Associations between the quantitative measurement of the FAZ and retinal VD measurements and outer retinal disruptions were evaluated.
The FAZ area was correlated with outer retinal layer disruption both in the superficial plexus (r = 0.244, 0.228, 0.212, P = 0.013, 0.02, 0.031 for the ELM, EZ, and IZ, respectively) and the deep capillary plexus (r = 0.298, 0.234, 0.197, P = 0.002, 0.019, 0.048 for the ELM, EZ, and IZ, respectively). A significant relationship was also found between the VD measurements in the SCP and DCP in ETDRS sectors and the outer retinal layers disruption.
The results of the current study show a significant relationship between the quantitative OCTA parameters and the integrity of the outer retinal layers. This finding reveals a correlation between retinal capillary nonperfusion and outer retinal disruption in eyes with diabetic retinopathy.

OBJECTIVE: A new clinical ultrahigh-resolution spectral domain optical coherence tomography (UHR-SD-OCT) system using an original averaging technique named \"A-scan matching algorithm\" was developed. The aim of this study was to determine whether our new UHR-SD-OCT system can obtain clearer sectional images of the retina than conventional standard resolution SD-OCT systems (SR-SD-OCT).
METHODS: We recorded horizontal B-scan images of 42 normal eyes using our new UHR-SD-OCT device (Bi-μ, Kowa) and a conventional SR-SD-OCT (Spectralis, Heidelberg). To evaluate the clarity of the interdigitation zone (IZ) subjectively, the integrity of IZ was divided into three types by two raters. To evaluate the clarity of the IZ objectively, a peak height score (PHS) was calculated at five different points of the macula using the longitudinal reflectivity profile.
RESULTS: The mean (± SD) of the subjective visibility score of the IZ in the UHR-SD-OCT images was 2.64 ± 0.54 which was significantly higher than the 2.46 ± 0.46 in the SR-SD-OCT images (P = 0.02). The PHS was also significantly higher for the UHR-SD-OCT than for the SR-SD-OCT images at all five locations (all P < 0.01).
CONCLUSIONS: The results indicate that the newly developed clinical UHR-SD-OCT instrument using the A-scan matching algorithm can obtain clearer images of the IZ, and they suggest that this device should be clinically useful in detecting finer structural abnormalities of the outer retina.