Facile synthesis of ultrabright luminogens with specific lipid droplets targeting feature for in vivo two-photon fluorescence retina imaging

The application of fluorescent probes for in vivo retinal imaging is of great importance,which could provide direct and crucial imaging evidence for a better understanding of common eye diseases.Herein,a group of bright organic luminogens with typical electron-donating(D)and electron-accepting(A)structures(abbreviated as LDs-BDM,LDs-BTM,and LDs-BHM)was synthesized through a simple single-step reaction.They were found to be efficient solid-state emitters with high fluorescence quantum yields of above 70%(e.g.,83.7%for LDs-BTM).Their light-emission properties could be tuned by the modulation ofπ-conjugation effect with methoxy groups at different substituent positions.Their resulting fluorescent nanoparticles(NPs)were demonstrated as specific lipid droplets(LDs)targeting probes with high brightness,good biocompatibility,and satisfactory photostability.LDs-BTM NPs with a large two-photon absorption cross section(σ2=249 GM)were further utilized as ultrabright two-photon fluorescence(2PF)nanoprobes for in vivo retina imaging of live zebrafish by NIR excitation at an ultralow concentration(0.5μmol/L).Integrated histological structures at the tissue level and corresponding fine details at the cellular level of the embryonic retina of live zebrafish were clearly demonstrated.This is the first report of using ultrabright LDs-targeting nanoprobes to accurately measure fine details in the retina with 2PF microscopic technique.These good results are anticipated to open up a new avenue in the development of efficient 2PF emitters for non-invasive bioimaging of living animals.

Bimorph deformable mirror based adaptive optics scanning laser ophthalmoscope for retina imaging in vivo

A bimorph deformable mirror （DM） with a large stroke of more than 30 μm using 35 actuators is presented and characterized for an adaptive optics （AO） confocal scanning laser ophthalmoscope application. Facilitated with a Shack-Hartmann wavefront sensor, the bimorph DM-based AO operates closed-loop AO corrections for hu- man eyes and reduces wavefront aberrations in most eyes to below 0.1 μm rms. Results from living eyes, including one exhibiting ~5D of myopia and ~2D of astigmatism along with notable high-order aberrations, reveal a prac- tical efficient aberration correction and demonstrate a great benefit for retina imaging, including improving resolution, increasing brightness, and enhancing the contrast of images.

Adaptive optics scanning laser ophthalmoscopy in fundus imaging, a review and update

Adaptive optics scanning laser ophthalmoscopy（AOSLO） has been a promising technique in funds imaging with growing popularity. This review firstly gives a brief history of adaptive optics（AO） and AO-SLO. Then it compares AO-SLO with conventional imaging methods（fundus fluorescein angiography, fundus autofluorescence, indocyanine green angiography and optical coherence tomography） and other AO techniques（adaptive optics flood-illumination ophthalmoscopy and adaptive optics optical coherence tomography）. Furthermore, an update of current research situation in AO-SLO is made based on different fundus structures as photoreceptors（cones and rods）, fundus vessels, retinal pigment epithelium layer, retinal nerve fiber layer, ganglion cell layer and lamina cribrosa. Finally, this review indicates possible research directions of AO-SLO in future.

Automatic Feature Extraction from Ocular Images

Ocular images processing is an important task in: i) biometrics system based on retina and/or sclera images, and ii) in clinical ophthalmology diagnosis of diseases like various vascular disorders. We presents a general framework for image processing of ocular images with a particular view on feature extraction. The method uses the set of geometrical and texture features and based on the information of the complex vessel structure of the retina and sclera. The feature extraction contains the image preprocessing, locating and segmentation of the region of interest (ROI). The image processing of ROI and the feature extraction are proceeded, and then the feature vector is determined for the human recognition and ophthalmology diagnosis.

Biophotons Contribute to Retinal Dark Noise

The discovery of dark noise in retinal photoreceptors resulted in a long-lasting controversy over its origin and the underlying mechanisms.Here,we used a novel ultra-weak biophoton imaging system（UBIS） to detect biophotonic activity（emission） under dark conditions in rat and bullfrog（Rana catesbeiana） retinas in vitro.We found a significant temperature-dependent increase in biophotonic activity that was completely blocked either by removing intracellular and extracellular Ca^（2＋）together or inhibiting phosphodiesterase 6.These findings suggest that the photon-like component of discrete dark noise may not be caused by a direct contribution of the thermal activation of rhodopsin,but rather by an indirect thermal induction of biophotonic activity,which then activates the retinal chromophore of rhodopsin.Therefore,this study suggests a possible solution regarding the thermal activation energy barrier for discrete dark noise,which has been debated for almost half a century.