![[Translate to chinese:] Raw widefield and THUNDER image of transversal mouse adult fiber lens section. Courtesy N. Houssin, Plagemen lab, Ohio State University, Columbus, USA. Transversal_mouse_adult_fiber_lens_section_Widefield_THUNDER_teaser.jpg](/fileadmin/_processed_/3/0/csm_Transversal_mouse_adult_fiber_lens_section_Widefield_THUNDER_teaser_e7400ac99b.jpg "[Translate to chinese:] Raw widefield and THUNDER image of transversal mouse adult fiber lens section. Courtesy N. Houssin, Plagemen lab, Ohio State University, Columbus, USA.")
研究眼睛形态发生
可将小鼠晶状体作为模型系统进行研究，以了解形态发生时间，例如，细胞形状变化、以及细胞迁移和增殖，这些对晶状体的形成都非常重要[1，2]。这些过程中断会导致形成出生缺陷，但是对这些中断的根源尚不了解[1，2]。科学家们结合使用小鼠遗传学、鸡胚学和细胞培养来打造模型系统，帮助其在细胞和分子层面阐明眼睛形态发生机制。
挑战
使用宽场显微镜观察厚样本时，由于光散射会产生模糊或离焦模糊，所以面临着巨大的挑战[3]。模糊可能会掩盖样本内部深处的目标结构。
方法
成像的样本是小鼠成人纤维晶状体的厚横向玻片。用抗ß连环蛋白抗体将BDB610153膜蛋白标记为绿色，用抗丝状肌动蛋白结合蛋白抗体将PA1-25076肌动蛋白丝结合蛋白标记为红色，并使用Hoechst将细胞核标记为蓝色。使用THUNDER Imager 3D Assay对该玻片成像并应用了large volume computational clearing（LVCC）[3，4]。
结果
小鼠晶状体玻片的图像如下面的图1所示。
![[Translate to chinese:] A) raw widefield image and B) image after LVCC of a transversal mouse adult fiber lens section. The images show the cell organization where green indicates BDB610153 membrane proteins, red Afadin actin filament-binding proteins, and blue nuclei.](/fileadmin/_processed_/5/0/csm_Transversal_mouse_adult_fiber_lens_section_Widefield_THUNDER_16c6426f5e.jpg "[Translate to chinese:] A) raw widefield image and B) image after LVCC of a transversal mouse adult fiber lens section. The images show the cell organization where green indicates BDB610153 membrane proteins, red Afadin actin filament-binding proteins, an")
结论
此处所示的结果证明，使用传统的宽场显微镜系统采集的小鼠纤维晶状体玻片图像非常模糊[3，4]，而使用THUNDER Imager 3D Assay时，展示目标区域的能力大大提高。用户通过它能够清晰地观察细节，即使是在完好的样本内部深处，也能实时进行观察，而不会出现任何的模糊。
References
- N.S. Houssin, J.B. Martin, V. Coppola, S. Ok Yoon, T.F. Plageman, Formation and contraction of multicellular actomyosin cables facilitate lens placode invagination, Developmental Biology (2020) vol. 462, iss. 1, pp. 36-49, DOI: 10.1016/j.ydbio.2020.02.014.
- B. Chauhan, T. Plageman, M. Lou, R. Lang, Epithelial Morphogenesis: The Mouse Eye as a Model System, Ch. 11 in Current Topics in Developmental Biology, Ed. P.A. Trainor (Academic Press, 2015), vol. 111, pp. 375-399, DOI: 10.1016/bs.ctdb.2014.11.011.
- J. Schumacher, L. Bertrand, Real time images of 3D specimens with sharp contrast free of haze: Technology Note THUNDER Imagers: How Do They Really Work? Science Lab (2019) Leica Microsystems.
- L. Felts, V. Kohli, J.M. Marr, J. Schumacher, O. Schlicker, An Introduction to Computational Clearing: A New Method to Remove Out-of-Focus Blur, Science Lab (2020) Leica Microsystems.
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