Adaptive Optical Two-Photon Fluorescence Microscopy Probes Cellular Organization of Ocular Lenses In Vivo

Paidi, Santosh Kumar; Zhang, Qinrong; Yang, Yuhan; Xia, Chun-Hong; Ji, Na; Gong, Xiaohua

doi:10.1167/iovs.64.7.20

Adaptive Optical Two-Photon Fluorescence Microscopy Probes Cellular Organization of Ocular Lenses In Vivo

Journal Article · Mon Jun 12 00:00:00 EDT 2023 · Investigative Opthalmology & Visual Science

DOI:https://doi.org/10.1167/iovs.64.7.20· OSTI ID:2470995

Paidi, Santosh Kumar ^[1]; Zhang, Qinrong ^[1]; Yang, Yuhan ^[1]; Xia, Chun-Hong ^[1]; Ji, Na ^[2]; Gong, Xiaohua ^[1]

University of California, Berkeley, CA (United States)
University of California, Berkeley, CA (United States); Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)

The mammalian ocular lens is an avascular multicellular organ that grows continuously throughout life. Traditionally, its cellular organization is investigated using dissected lenses, which eliminates in vivo environmental and structural support. Therefore, in vivo optical imaging methods for studying lenses in their native context in live animals are urgently needed. Here, we demonstrated that two-photon fluorescence microscopy can visualize lens cells in vivo. To maintain subcellular resolution at depth, we used adaptive optics to correct aberrations owing to ocular and lens tissues, which led to substantial signal and resolution improvements. Imaging lens cells up to 980 µm deep, we observed novel cellular organizations including suture-associated voids, enlarged vacuoles, and large cavities, contrary to the conventional view of a highly ordered organization. We tracked these features longitudinally over weeks and observed the incorporation of new cells during growth. Taken together, noninvasive longitudinal in vivo imaging of lens morphology using adaptive optics two-photon fluorescence microscopy will allow us to observe the development or alterations of lens cellular organization in living animals directly.

View Accepted Manuscript (DOE)

Research Organization:: Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)

Sponsoring Organization:: USDOE

Grant/Contract Number:: AC02-05CH11231

OSTI ID:: 2470995

Journal Information:: Investigative Opthalmology & Visual Science, Journal Name: Investigative Opthalmology & Visual Science Journal Issue: 7 Vol. 64; ISSN 1552-5783

Publisher:: Association for Research in Vision and OphthalmologyCopyright Statement

Country of Publication:: United States

Language:: English

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