PEGDA-based HistoBrick for increasing throughput of cryosectioning and immunohistochemistry in organoid and small tissue studies.

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dc.contributor.authorVuille-Dit-Bille, Emilie
dc.contributor.authorUtz, Larissa
dc.contributor.authorMüllner, Fiona E
dc.contributor.authorArteaga-Moreta, Valeria J
dc.contributor.authorHou, Yanyan
dc.contributor.authorSpirig, Stefan E
dc.contributor.authorLedroit-Paic, Diane
dc.contributor.authorHeub, Sarah
dc.contributor.authorGoldowsky, Jonas
dc.contributor.authorWeder, Gilles
dc.contributor.authorRenner, Magdalena
dc.date.accessioned2025-04-10T13:35:59Z
dc.date.available2025-04-10T13:35:59Z
dc.date.issued2025-01-02
dc.description.abstractHistology is the gold standard for analyzing tissue structure and cell morphology. Immunostaining on thin tissue sections enables precise visualization of antigens and proteins. However, for cryosectioning small tissues such as organoids, spheroids, and tumoroids there is a lack of standardized, time- and cost-effective methods, limiting the throughput of analysis. Here, we have adapted to cryosectioning our previously developed HistoBrick approach, in which small tissue arrangement is spatially controlled within arrayed mini-wells. By testing various embedding matrices, we show that an 8% PEGDA and 2.5% gelatine mixture is optimal, providing essential structural support to maintain sample integrity during cryosectioning. This embedding matrix preserves fragile substructures of human retinal organoids, which are particularly susceptible to damage during sample preparation. Using PEGDA-gelatine HistoBricks for the simultaneous embedding of 16 retinal organoids, we analyzed a time course of retinal organoid development. We observed the maintenance of photoreceptors cell bodies up to week 98 in culture, while photoreceptor outer segments were gradually lost. Further, we observed displaced photoreceptors in the region of outer segments. The PEGDA-gelatine HistoBrick is a cost-efficient tool that can be implemented for small tissue studies to increase throughput in experiments such as large-scale screenings or toxicology research.
dc.description.sponsorshipResearch Support, Non-U.S. Gov't
dc.identifier.citationVuille-dit-Bille, E., Utz, L., Müllner, F.E. et al. PEGDA-based HistoBrick for increasing throughput of cryosectioning and immunohistochemistry in organoid and small tissue studies. Sci Rep 15, 412 (2025). https://doi.org/10.1038/s41598-024-83164-2
dc.identifier.doi10.1038/s41598-024-83164-2
dc.identifier.issn2045-2322
dc.identifier.other39747958
dc.identifier.pii10.1038/s41598-024-83164-2
dc.identifier.pmid39747958
dc.identifier.urihttps://hdl.handle.net/20.500.12839/1654
dc.language.isoen
dc.rights© 2024. The Author(s).
dc.rightsAttribution-NonCommercial-NoDerivatives 4.0 International*
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/4.0/*
dc.source.beginpage412
dc.source.countryEngland
dc.source.issue1
dc.source.journaltitleScientific reports
dc.source.volume15
dc.subject3D cell models
dc.subjectCryosection
dc.subjectHistoBrick
dc.subjectHistology
dc.subjectHuman retinal organoids
dc.subjectMicrotissues
dc.subjectOrganoids
dc.subjectSpheroids
dc.titlePEGDA-based HistoBrick for increasing throughput of cryosectioning and immunohistochemistry in organoid and small tissue studies.
dc.typeJournal Article
dc.type.csemdivisionsBU-R
dc.type.csemresearchareasTools for Life Sciences
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