Quantitative Analysis of Whole-Mount Fluorescence-Stained Tumor Spheroids in Phenotypic Drug Screens.

3D cell culture Image analysis Open source Optical tissue clearing Organoids Segmentation Spheroid

Journal

Methods in molecular biology (Clifton, N.J.)
ISSN: 1940-6029
Titre abrégé: Methods Mol Biol
Pays: United States
ID NLM: 9214969

Informations de publication

Date de publication:
2024
Historique:
medline: 23 2 2024
pubmed: 23 2 2024
entrez: 23 2 2024
Statut: ppublish

Résumé

Three-dimensional cell cultures, such as spheroids or organoids, serve as important models for drug screening purposes. Optical tissue clearing (OTC) enhances the visualization of fluorescence stainings and enables in toto microscopy of 3D cell culture models. Furthermore, subsequent automated image analysis tools convert qualitative confocal image sets into quantitative data. In this chapter, we describe a detailed protocol for preparation of HT29 cancer spheroids, 3D in toto immunostaining, glycerol-based OTC, whole-mount imaging, and semi-automated downstream image processing and segmentation for nuclear image analysis using open-source software.

Identifiants

DOI: 10.1007/978-1-0716-3674-9_20 PMID: 38393603
pubmed: 38393603
doi: 10.1007/978-1-0716-3674-9_20
doi:

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

311-334

Informations de copyright

© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.

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Auteurs

Elina Nuernberg (E)

Institute of Molecular and Cell Biology, Mannheim University of Applied Sciences, Mannheim, Germany.
Center for Mass Spectrometry and Optical Spectroscopy (CeMOS), Mannheim University of Applied Sciences, Mannheim, Germany.

Roman Bruch (R)

Institute of Molecular and Cell Biology, Mannheim University of Applied Sciences, Mannheim, Germany.
Institute for Automation and Applied Informatics, Karlsruhe Institute of Technology (KIT), Eggenstein-Leopoldshafen, Germany.

Mathias Hafner (M)

Institute of Molecular and Cell Biology, Mannheim University of Applied Sciences, Mannheim, Germany.

Ruediger Rudolf (R)

Institute of Molecular and Cell Biology, Mannheim University of Applied Sciences, Mannheim, Germany.
Center for Mass Spectrometry and Optical Spectroscopy (CeMOS), Mannheim University of Applied Sciences, Mannheim, Germany.

Mario Vitacolonna (M)

Institute of Molecular and Cell Biology, Mannheim University of Applied Sciences, Mannheim, Germany. m.vitacolonna@hs-mannheim.de.
Center for Mass Spectrometry and Optical Spectroscopy (CeMOS), Mannheim University of Applied Sciences, Mannheim, Germany. m.vitacolonna@hs-mannheim.de.

Classifications MeSH