An efficient and user-friendly method for cytohistological analysis of organoids

一种高效且用户友好的类器官细胞组织学分析方法

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作者:Shi-Wei Zhang, Wei Chen, Xiao-Fang Lu, Zhang Wen, Lei Hu, Yu-Hong Liu, Zheng Yang, Ling Xue, Qiao Su, Le-Ping Yan, Joaquim M Oliveira, Rui L Reis, Yu-Long He, Chang-Hua Zhang

Abstract

Organoid culture is a recently developed in vitro three-dimensional (3D) cell culture technology. It has wide applications in tissue engineering studies. However, histological analysis of organoid is quite complex and tedious for researchers. This study proposes a user-friendly, affordable and efficient method for making formalin-fixed paraffin embedded (FFPE) organoid blocks and Optimal Cutting Temperature compound (OCT) embedded frozen organoid blocks. This method implements a key pre-embedding step for preparing paraffin embedded organoid blocks, which could concentrate organoid together without damaging or loss of samples. This method could be used to process even a small number of organoids with high efficiency. In addition, with minor modifications, the method is readily applied for OCT embedded organoid blocks. The slides generated were ready for H&E staining, immunohistochemistry staining and immunofluorescent staining. The method described in this study can be easily used for routine histological analysis of organoid, and could be performed in general pathology labs and requires no dedicated equipment and reagent.

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