Light Sheet Fluorescence Microscopy Eye Exam

Following iDISCO+-based clearing and immunolabeling, researchers devised a protocol for the complete human eye’s light sheet fluorescence microscopy (LSFM). En-face observation of the fundus and resolution of retinal capillaries are made possible by this method’s ability to provide three-dimensional (3D) navigation and personalized presentation. This technique is helpful in anatomopathology and research because it gets over the drawbacks of conventional histology and makes monitoring spatially complicated structures possible.

Light sheet fluorescence microscopy (LSFM), which offers faster acquisition times than confocal microscopy, has recently been developed to enable volumetric imaging of immunolabeled cleared materials. Large, spatially complex structures may be thoroughly analyzed with this method, allowing for quicker reconstruction than with standard histology. Clearing protocols like CLARITY, BABB, 3DISCO, and CUBIC are often used.

Ocular histopathology is particularly difficult because of the eye’s spherical shape and varied makeup. Eliminating ocular tissues may provide a sophisticated approach to histology. There have been few studies on ocular tissue cleaning, though. Recently, LSFM of whole murine eyes was attained by researchers, allowing for the time course of vascular development to be documented and hyaloid vessel organization to be observed.

Through enhanced bleaching techniques, linearization to promote antibody penetration, and a modified iDISCO+ clearing methodology, researchers have accomplished LSFM of complete immunolabelled human eyeballs. A specially-built mesoSPIM and thorough numerical dissection enable multiscale, tailored imaging of eye tissues, including pseudo-fundoscopic (en face) vision. Because of conventional techniques, clearing the entire human eye—a sphere of 6 cm3—has proven difficult.

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