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Quantitation of mitotic cells in the neural tube of zebrafish embryos using automated nuclei counting

Here, we provide a protocol to automate the quantification of the number of phospho-histone H3-positive cells in the developing nervous system of zebrafish using a custom MATLAB script to identify labeled nuclei. We describe steps for fixation, immunolabeling, and imaging of zebrafish embryos. We th...

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Detalles Bibliográficos
Autores principales: Emond, Michelle R., Jontes, James D.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10519849/
https://www.ncbi.nlm.nih.gov/pubmed/37738116
http://dx.doi.org/10.1016/j.xpro.2023.102596
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author Emond, Michelle R.
Jontes, James D.
author_facet Emond, Michelle R.
Jontes, James D.
author_sort Emond, Michelle R.
collection PubMed
description Here, we provide a protocol to automate the quantification of the number of phospho-histone H3-positive cells in the developing nervous system of zebrafish using a custom MATLAB script to identify labeled nuclei. We describe steps for fixation, immunolabeling, and imaging of zebrafish embryos. We then detail the analysis steps using Fiji and MATLAB. This protocol can be used for fixed, immunolabeled tissue, as shown here, or for live samples, such as cells expressing a histone-GFP fusion protein. For complete details on the use and execution of this protocol, please refer to Biswas et al.(1)
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spelling pubmed-105198492023-09-27 Quantitation of mitotic cells in the neural tube of zebrafish embryos using automated nuclei counting Emond, Michelle R. Jontes, James D. STAR Protoc Protocol Here, we provide a protocol to automate the quantification of the number of phospho-histone H3-positive cells in the developing nervous system of zebrafish using a custom MATLAB script to identify labeled nuclei. We describe steps for fixation, immunolabeling, and imaging of zebrafish embryos. We then detail the analysis steps using Fiji and MATLAB. This protocol can be used for fixed, immunolabeled tissue, as shown here, or for live samples, such as cells expressing a histone-GFP fusion protein. For complete details on the use and execution of this protocol, please refer to Biswas et al.(1) Elsevier 2023-09-21 /pmc/articles/PMC10519849/ /pubmed/37738116 http://dx.doi.org/10.1016/j.xpro.2023.102596 Text en © 2023 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Emond, Michelle R.
Jontes, James D.
Quantitation of mitotic cells in the neural tube of zebrafish embryos using automated nuclei counting
title Quantitation of mitotic cells in the neural tube of zebrafish embryos using automated nuclei counting
title_full Quantitation of mitotic cells in the neural tube of zebrafish embryos using automated nuclei counting
title_fullStr Quantitation of mitotic cells in the neural tube of zebrafish embryos using automated nuclei counting
title_full_unstemmed Quantitation of mitotic cells in the neural tube of zebrafish embryos using automated nuclei counting
title_short Quantitation of mitotic cells in the neural tube of zebrafish embryos using automated nuclei counting
title_sort quantitation of mitotic cells in the neural tube of zebrafish embryos using automated nuclei counting
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10519849/
https://www.ncbi.nlm.nih.gov/pubmed/37738116
http://dx.doi.org/10.1016/j.xpro.2023.102596
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