State Key Laboratory for Animal Disease Control and Prevention, Key Laboratory of Veterinary Public Health of Agriculture Ministry Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China.
Int J Mol Sci. 2023 Sep 1;24(17):13568. doi: 10.3390/ijms241713568.
Intestinal organoids have emerged as powerful model systems for studying the complex structure and function of the intestine. However, there is a lack of widely applicable methods for the collection, labeling, and imaging of intestinal organoids. In this study, we developed a novel method for loading and labeling intestinal organoids, a method that efficiently collects the organoids and facilitates imaging of their three-dimensional (3D) structure. Based on this strainer platform, mouse intestinal organoids were adequately collected and immobilized, facilitating the immunolabeling workflow to target proteins of the organoids. After evaluation, the strainer size of 40 μm was considered to be more conducive to the collection and labeling of mouse intestinal organoids. More extensive research on organoids of multiple types and species origins will contribute to broadening the applicability of the methodology. Overall, our study proposes an innovative workflow for loading and analyzing intestinal organoids. The combination of a strainer-based collection method, fluorescent labeling, and 3D reconstruction provides valuable insights into the organization and complexity of these tissue models, thereby offering new avenues for investigating intestinal development, disease modeling, and drug discovery.
肠类器官已成为研究肠道复杂结构和功能的强大模型系统。然而，目前缺乏广泛适用于肠类器官收集、标记和成像的方法。在本研究中，我们开发了一种新型的肠类器官加载和标记方法，该方法能够高效地收集类器官，并方便对其三维（3D）结构进行成像。基于这个滤网平台，成功收集并固定了小鼠肠类器官，便于对类器官的目标蛋白进行免疫标记工作流程。经过评估，40 μm 的滤网尺寸更有利于收集和标记小鼠肠类器官。对多种类型和物种来源的类器官进行更广泛的研究将有助于扩大该方法的适用性。总的来说，本研究提出了一种用于加载和分析肠类器官的创新工作流程。基于滤网的收集方法、荧光标记和 3D 重建的结合，为这些组织模型的组织和复杂性提供了有价值的见解，从而为研究肠道发育、疾病建模和药物发现开辟了新途径。
