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  Preparing Planarian Cells for High-Content Fluorescence Microscopy Using RNA in Situ Hybridization and Immunocytochemistry

Grohme, M. A., Frank, O., & Rink, J. C. (2023). Preparing Planarian Cells for High-Content Fluorescence Microscopy Using RNA in Situ Hybridization and Immunocytochemistry. In L. Gentile (Ed.), Schmidtea Mediterranea: Methods and Protocols (pp. 121-155). New York, NY: Springer US. doi:10.1007/978-1-0716-3275-8_8.

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Item Permalink: https://hdl.handle.net/21.11116/0000-000E-06D4-C Version Permalink: https://hdl.handle.net/21.11116/0000-000E-06D5-B
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978-1-0716-3275-8_8.pdf (Publisher version), 520KB
 
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 Creators:
Grohme, Markus A., Author
Frank, Olga, Author
Rink, Jochen C.1, Author           
Affiliations:
1Department of Tissue Dynamics and Regeneration, Max Planck Institute for Multidisciplinary Sciences, Max Planck Society, ou_3350274              

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 Abstract: High-content fluorescence microscopy combines the efficiency of high-throughput techniques with the ability to extract quantitative information from biological systems. Here we describe a modular collection of assays adapted for fixed planarian cells that enable multiplexed measurements of biomarkers in microwell plates. These include protocols for RNA fluorescent in situ hybridization (RNA FISH) as well as immunocytochemical protocols for quantifying proliferating cells targeting phosphorylated histone H3 as well as 5-bromo-2′-deoxyuridine (BrdU) incorporated into the nuclear DNA. The assays are compatible with planarians of virtually any size, as the tissue is disaggregated into a single-cell suspension before fixation and staining. By sharing many reagents with established planarian whole-mount staining protocols, preparation of samples for high-content microscopy adoption requires little additional investment.

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Language(s): eng - English
 Dates: 2023-07-112023
 Publication Status: Issued
 Pages: -
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 Table of Contents: -
 Rev. Type: Peer
 Identifiers: DOI: 10.1007/978-1-0716-3275-8_8
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Title: Schmidtea Mediterranea: Methods and Protocols
Source Genre: Book
 Creator(s):
Gentile, Luca, Editor
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Publ. Info: New York, NY : Springer US
Pages: - Volume / Issue: 2680 Sequence Number: - Start / End Page: 121 - 155 Identifier: ISSN: 1064-3745
ISSN: 1940-6029
ISBN: 978-1-0716-3274-1
ISBN: 978-1-0716-3275-8

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Title: Methods in Molecular Biology
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Pages: - Volume / Issue: 2680 Sequence Number: - Start / End Page: - Identifier: -