School of Optometry, University of California, Berkeley, California, United States.
Department of Physics, University of California, Berkeley, California, United States.
Invest Ophthalmol Vis Sci. 2023 Jun 1;64(7):20. doi: 10.1167/iovs.64.7.20.
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.
哺乳动物眼球晶状体是一个无血管的多细胞器官，在整个生命过程中不断生长。传统上，通过解剖晶状体来研究其细胞组织，这消除了体内的环境和结构支持。因此，迫切需要在活体动物的天然环境中研究晶状体的体内光学成像方法。
在这里，我们证明了双光子荧光显微镜可以在体内可视化晶状体细胞。为了在深度上保持亚细胞分辨率，我们使用自适应光学来校正由于眼部和晶状体组织引起的像差，这导致了信号和分辨率的显著提高。
对深度达 980µm 的晶状体细胞进行成像，我们观察到了一些新的细胞组织，包括缝线相关的空隙、扩大的空泡和大腔，这与传统的高度有序组织的观点相悖。我们对这些特征进行了数周的纵向跟踪，并在生长过程中观察到了新细胞的掺入。
综上所述，使用自适应光学双光子荧光显微镜对晶状体形态进行非侵入性的纵向体内成像，将使我们能够直接观察活体动物中晶状体细胞组织的发育或改变。
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