Advancements in 3D spheroid imaging: Optimised cryosectioning and immunostaining techniques.
| Autor: | Charlet-Faure C; Department of Biochemistry and Molecular Biology, University of Southern Denmark, Campusvej 55, Odense M DK-5230, Denmark.; Faculté des Sciences Site St Charles Aix Marseille Université, 3 place Victor Hugo, Marseille 13331, France., Thulesen AP; Department of Biochemistry and Molecular Biology, University of Southern Denmark, Campusvej 55, Odense M DK-5230, Denmark., Rogowska-Wrzesinska A; Department of Biochemistry and Molecular Biology, University of Southern Denmark, Campusvej 55, Odense M DK-5230, Denmark. |
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| Jazyk: | angličtina |
| Zdroj: | MethodsX [MethodsX] 2023 Oct 06; Vol. 11, pp. 102415. Date of Electronic Publication: 2023 Oct 06 (Print Publication: 2023). |
| DOI: | 10.1016/j.mex.2023.102415 |
| Abstrakt: | This article presents a modified protocol for embedding and sectioning spheroids and organoids, which are increasingly used in research due to their ability to emulate living tissue. The modifications aim to reduce the distortion and damage of these fragile structures during the embedding and sectioning process. The new method involves using optimized embedding containers, a modified embedding protocol, and optimized temperatures for cryosectioning. A heat-induced antigen retrieval protocol was tested and found to significantly increase immunostaining intensity without compromising spheroid integrity. The combined approach allowed for the creation of thinner cryosections, leading to clearer and more detailed images. The results suggest that the modified protocol could be widely adopted to enhance the imaging of spheroids and organoids.•Paraformaldehyde fixation of spheroids•Antigen retrieval treatment of spheroids•Embedding in freezing medium and cryosectioning. Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. (© 2023 The Authors. Published by Elsevier B.V.) |
| Databáze: | MEDLINE |
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