Cargando…
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...
Autores principales: | , |
---|---|
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 |
_version_ | 1785109784472584192 |
---|---|
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) |
format | Online Article Text |
id | pubmed-10519849 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT emondmicheller quantitationofmitoticcellsintheneuraltubeofzebrafishembryosusingautomatednucleicounting AT jontesjamesd quantitationofmitoticcellsintheneuraltubeofzebrafishembryosusingautomatednucleicounting |