Lallemant Louison, Lebreton Corinne, Garfa-Traoré Meriem
Cell Imaging Platform, INSERM-US24-CNRS UMS 3633 Structure Fédérative de Recherche Necker, Paris University, Paris 75015, France.
Université de Paris, Imagine Institute, Laboratory of Intestinal Immunity, INSERM UMR1163, Paris 75015, France.
J Biol Methods. 2020 Dec 28;7(4):e141. doi: 10.14440/jbm.2020.334. eCollection 2020.
An organoid is a three-dimensional multicellular structure that shows realistic micro-anatomy of an organ. This model mimics the environment, architecture and multi-lineage differentiation of the original organs and allows to answer many interesting biological questions. For these reasons, they are widely used in stem cell, regenerative medicine, toxicology, pharmacology, and host-microbe interactions research. In order to study organoids, microscopy is very useful: It is possible to make three-dimensional reconstruction of serial sections but it is time consuming and error-prone. Here we propose an alternative solution: Tissue clearing reduces the dispersion of light because it homogenizes the refractive index of the tissue, allowing sample observation throughout its thickness. We have compared different clearing techniques on mouse intestinal organoids using different acquisition methods.
类器官是一种三维多细胞结构，展现出器官逼真的微观解剖结构。该模型模拟了原始器官的环境、结构和多谱系分化，能够回答许多有趣的生物学问题。基于这些原因，它们被广泛应用于干细胞、再生医学、毒理学、药理学以及宿主-微生物相互作用研究。为了研究类器官，显微镜检查非常有用：可以对连续切片进行三维重建，但这既耗时又容易出错。在此，我们提出一种替代解决方案：组织透明化可减少光的散射，因为它使组织的折射率均匀化，从而能够对整个样本厚度进行观察。我们使用不同的采集方法，比较了小鼠肠道类器官上的不同透明化技术。
