Authors
Jiawei Li, Rong Jin, Xuanxuan Zhang, Yuecong Chen, Huihan Ai, Xianglin Mei, Ying Sun, Lihua Zheng, Guannan Wang, Yongli Bao, Xiaoli Li
Published in
MethodsX. Volume 17. Pages 104016. Epub Jun 25, 2026.
Abstract
Mouse intestinal organoids are essential 3D models that retain host genetic characteristics and complex architecture, making them invaluable for intestinal biology research. However, traditional cultivation and histological processing often suffer from high operational complexity and sample loss during sectioning. This protocol provides an optimized workflow for the establishment of mouse intestinal organoid cultures and their subsequent preparation for immunohistochemistry and fluorescence staining. By refining the handling steps, the method significantly reduces total operational time while enhancing experimental quality. Furthermore, the protocol addresses common challenges in structural integrity, ensuring that the delicate crypt-villus morphology remains intact during the sectioning process. This comprehensive approach offers a robust framework for high-quality histological analysis, supported by a comparative evaluation with traditional methods to guide researchers in different experimental contexts.•Introduces a direct pellet-OCT embedding strategy that minimizes organoid transfer and sample loss during histological processing.•Enables high-quality frozen sections compatible with H&E, Alcian Blue, immunohistochemistry, and immunofluorescence staining.•Provides a practical alternative to conventional agarose-paraffin workflows while preserving organoid morphology and reducing processing complexity.
PMID:
42405229
Bibliographic data and abstract were imported from PubMed on 06 Jul 2026.
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