Adaptive optical two-photon fluorescence microscopy probes cellular organization of ocular lenses in vivo.
Santosh Kumar PaidiQinrong ZhangYuhan YangChun-Hong XiaNa JiXiaohua GongPublished in: bioRxiv : the preprint server for biology (2023)
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. Here, we demonstrated that two-photon fluorescence microscopy (2PFM) can visualize lens cells in vivo. To maintain subcellular resolution at depth, we employed adaptive optics (AO) to correct aberrations due 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, non-invasive longitudinal in vivo imaging of lens morphology using AO 2PFM will allow us to directly observe the development or alterations of lens cellular organization in living animals.
Keyphrases
- single molecule
- induced apoptosis
- high resolution
- cataract surgery
- cell cycle arrest
- living cells
- optical coherence tomography
- cell death
- small molecule
- gene expression
- endoplasmic reticulum stress
- oxidative stress
- signaling pathway
- high throughput
- cell proliferation
- optic nerve
- mass spectrometry
- preterm birth
- label free
- monte carlo