Journal
JOURNAL OF TISSUE ENGINEERING AND REGENERATIVE MEDICINE
Volume 15, Issue 11, Pages 1012-1022Publisher
WILEY
DOI: 10.1002/term.3248
Keywords
FFPE; immunofluorescence; immunohistochemistry; OCT; organoids analysis
Categories
Funding
- Shenzhen Fundamental Research Program [JCYJ20200109142605909]
- Sanming Project of Medicine in Shenzhen [SZSM201911010]
- Shenzhen Key Medical Discipline Construction Fund [SZXK016]
- Sun Yat-sen University [ZSQYRSFPD0008]
- Seventh Affiliated Hospital Sun Yat-sen University [ZSQYRSF0008]
- Science and Technology Planning Project of Guangdong Province [2019A030317005]
- Guangdong Provincial Key Laboratory of Digestive Cancer Research [2021B1212040006]
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This study introduces a user-friendly, cost-effective method for making FFPE and OCT embedded organoid blocks, facilitating histological analysis for researchers with no specialized equipment required.
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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