Establishment of a novel protocol for formalin-fixed paraffin embedded organoids and spheroids

• Published in: Biology open Vol. 12; no. 5
• Main Authors: Yoshimoto, Shohei, Taguchi, Masahide, Sumi, Satoko, Oka, Kyoko, Okamura, Kazuhiko
• Format: Journal Article
• Language: English
• Published: England The Company of Biologists 01.05.2023
• Subjects: formalin-fixed paraffin-embedded (ffpe); immunohistochemistry; organoid; spheroid; three-dimensional experimental model; Immunohistochemistry; Organoid; Spheroid; Formalin-fixed paraffin-embedded (FFPE); Three-dimensional experimental model
• ISSN: 2046-6390; 2046-6390
• DOI: 10.1242/bio.059882

Three-dimensional (3D) cell culture models such as spheroids and organoids are widely use in the field of experimental biology. To analyze these 3D experimental models, formalin-fixed paraffin-embedded (FFPE) sections are superior to whole-mount imaging in some experimental purposes, such as exploring samples with a depth limitation of primary antibody penetration immunohistochemically. However, tiny 3D cell culture samples are difficult to embed in paraffin and acquire appropriate sections. In this report, we optimized a protocol of paraffin embedding for spheroids and organoids. In addition, we compared FFPE sections with frozen sections in ratio of sample collection and section condition after staining, and could reproduce improved results reliably. The protocol we established could be widely used in many laboratories and become a useful technique for analyzing spheroids and organoids.