Minimum intensity projection of embossed quadrant-detection images for improved photoreceptor mosaic visualisation

Abstract

Non-confocal split-detection imaging reveals the cone photoreceptor inner segment mosaic in a plethora of retinal conditions, with the potential of providing insight to ageing, disease, and response to treatment processes, in vivo, and allows the screening of candidates for cell rescue therapies. This imaging modality complements confocal reflectance adaptive optics scanning light ophthalmoscopy, which relies on the waveguiding properties of cones, as well as their orientation toward the pupil. Split-detection contrast, however, is directional, with each cone inner segment appearing as opposite dark and bright semicircles, presenting a challenge for either manual or automated cell identification. Quadrant-detection imaging, an evolution of split detection, could be used to generate images without directional dependence. Here, we demonstrate how the embossed-filtered quadrant-detection images, originally proposed by Migacz et al. for visualising hyalocytes, can also be used to generate photoreceptor mosaic images with better and non-directional contrast for improved visualisation. As a surrogate of visualisation improvement between legacy split-detection images and the images resulting from the method described herein, we provide preliminary results of simple image processing routines that may enable the automated identification of generic image features, as opposed to complex algorithms developed specifically for photoreceptor identification, in pathological retinas.

Adaptive Optics Retinal Imaging in RDH12-Associated Early Onset Severe Retinal Dystrophy

“… 30 Confocal reflectance imaging revealed photoreceptor outer segments using a 790 nm superluminescent diode (Superlum, Cork, Ireland) whereas nonconfocal multiple-scattered light revealed photoreceptor inner segments using either a split- 17 or a quadrant-detection setup. 31 The light source for wavefront-sensing was an 850 nm superluminescent diode (Superlum), and the wavefront correction was performed by a Hi-Speed DM97-15 deformable mirror (Alpao, Montbonnot-Saint-Martin, France). Patient's pupils were dilated using a drop of 1% tropicamide and 2.5% phenylephrine each.…”

“…These four split directions were further processed using the emboss (edge-enhancing) filter in Photoshop CS6 (Adobe Systems, San Jose, CA, USA). 31 The embossed quadrant-detection image was the primary modality used for cell annotations (where available) because it allowed better resolution of both waveguiding and nonwaveguiding cells, compared with confocal or split-detection modalities, respectively.…”