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用于小鼠肠道组织光学透明化的快速高效方案，以增强荧光成像和三维重建。

Rapid and efficient protocol for optical clearing of mouse intestinal tissues for enhanced fluorescence imaging and 3D reconstruction.

作者信息

George Joel Johnson, Ball Sierra H, Wang Elena, Schneider Remy T, Capdevila Claudia, Martini Francesca, Lee Hyeonjeong, Miller Jonathan, Cheng Liang, Murray John W, Snoeck Hans-Willem, Yan Kelley S

机构信息

Department of Medicine, Columbia University Irving Medical Center, New York, NY, USA; Department of Genetics & Development, Columbia University Irving Medical Center, New York, NY, USA; Herbert Irving Comprehensive Cancer Center, Columbia University Irving Medical Center, New York, NY, USA; Digestive & Liver Diseases Research Center, Columbia University Irving Medical Center, New York, NY, USA; Columbia Center for Human Development/Center for Stem Cell Therapies, Columbia University Vagelos College of Physicians and Surgeons, New York, NY 10032, USA.

Department of Medicine, Columbia University Irving Medical Center, New York, NY, USA; Department of Genetics & Development, Columbia University Irving Medical Center, New York, NY, USA; Herbert Irving Comprehensive Cancer Center, Columbia University Irving Medical Center, New York, NY, USA; Digestive & Liver Diseases Research Center, Columbia University Irving Medical Center, New York, NY, USA; Columbia Center for Human Development/Center for Stem Cell Therapies, Columbia University Vagelos College of Physicians and Surgeons, New York, NY 10032, USA.

出版信息

STAR Protoc. 2025 Jun 20;6(2):103841. doi: 10.1016/j.xpro.2025.103841. Epub 2025 May 28.

DOI:10.1016/j.xpro.2025.103841
PMID:40440175
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC12156166/
Abstract

Histological analysis of intestinal epithelial tissues is enhanced by 3D visualization compared to 2D sections. Here, we present a protocol for 3D visualization of intestinal epithelial cells using an optical clearing approach optimized for endogenous fluorescence and preservation of crypt-villus morphology. We describe steps for clearing and refractive index matching tissue. We provide detailed procedures for imaging and reconstructing tissue to visualize epithelial cells along the crypt-villus axis with high resolution. We illustrate this approach with endogenous tdTomato used for lineage tracing in the small intestine of Fgfbp1-CreER; Rosa26-tdTomato mice. For complete details on the use and execution of this protocol, please refer to Capdevila et al..

摘要

与二维切片相比，三维可视化增强了肠道上皮组织的组织学分析。在此，我们展示了一种使用光学透明方法对肠道上皮细胞进行三维可视化的方案，该方法针对内源性荧光和隐窝 - 绒毛形态的保存进行了优化。我们描述了组织透明和折射率匹配的步骤。我们提供了成像和重建组织的详细程序，以高分辨率可视化沿隐窝 - 绒毛轴的上皮细胞。我们用用于Fgfbp1 - CreER; Rosa26 - tdTomato小鼠小肠谱系追踪的内源性tdTomato说明了这种方法。有关此方案的使用和执行的完整详细信息，请参考卡德维拉等人的研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4a96/12156166/11e392ec1242/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4a96/12156166/5762fdae0046/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4a96/12156166/44f39f1363b2/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4a96/12156166/11e392ec1242/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4a96/12156166/5762fdae0046/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4a96/12156166/44f39f1363b2/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4a96/12156166/11e392ec1242/gr2.jpg

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