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Brief freezing steps lead to robust immunofluorescence in the Drosophila nervous system
Drosophila melanogaster possesses a complex nervous system, regulating sophisticated behavioral outputs, that serves as a powerful model for dissecting molecular mechanisms underlying neuronal function and neurodegenerative disease. Immunofluorescence techniques provide a way to visualize the spatio...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Future Science Ltd
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7031821/ https://www.ncbi.nlm.nih.gov/pubmed/31687836 http://dx.doi.org/10.2144/btn-2018-0067 |
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author | Thapa, Ada Sullivan, Shea M Nguyen, Minh Q Buckley, Dominic Ngo, Vy T Dada, Austin O Blankinship, Essence Cloud, Veronica Mohan, Ryan D |
author_facet | Thapa, Ada Sullivan, Shea M Nguyen, Minh Q Buckley, Dominic Ngo, Vy T Dada, Austin O Blankinship, Essence Cloud, Veronica Mohan, Ryan D |
author_sort | Thapa, Ada |
collection | PubMed |
description | Drosophila melanogaster possesses a complex nervous system, regulating sophisticated behavioral outputs, that serves as a powerful model for dissecting molecular mechanisms underlying neuronal function and neurodegenerative disease. Immunofluorescence techniques provide a way to visualize the spatiotemporal organization of these networks, permitting observation of their development, functional location, remodeling and, eventually, degradation. However, basic immunostaining techniques do not always result in efficient antibody penetration through the brain, and supplemental techniques to enhance permeability can compromise structural integrity, altering spatial organization. Here, slow freezing of brains is shown to facilitate antibody permeability without loss of antibody specificity or brain integrity. To demonstrate the advantages of this freezing technique, the results of two commonly used permeation methods – detergent-based and partial proteolytic digestion – are compared. |
format | Online Article Text |
id | pubmed-7031821 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Future Science Ltd |
record_format | MEDLINE/PubMed |
spelling | pubmed-70318212020-03-03 Brief freezing steps lead to robust immunofluorescence in the Drosophila nervous system Thapa, Ada Sullivan, Shea M Nguyen, Minh Q Buckley, Dominic Ngo, Vy T Dada, Austin O Blankinship, Essence Cloud, Veronica Mohan, Ryan D Biotechniques Benchmark Drosophila melanogaster possesses a complex nervous system, regulating sophisticated behavioral outputs, that serves as a powerful model for dissecting molecular mechanisms underlying neuronal function and neurodegenerative disease. Immunofluorescence techniques provide a way to visualize the spatiotemporal organization of these networks, permitting observation of their development, functional location, remodeling and, eventually, degradation. However, basic immunostaining techniques do not always result in efficient antibody penetration through the brain, and supplemental techniques to enhance permeability can compromise structural integrity, altering spatial organization. Here, slow freezing of brains is shown to facilitate antibody permeability without loss of antibody specificity or brain integrity. To demonstrate the advantages of this freezing technique, the results of two commonly used permeation methods – detergent-based and partial proteolytic digestion – are compared. Future Science Ltd 2019-11-05 2019-12 /pmc/articles/PMC7031821/ /pubmed/31687836 http://dx.doi.org/10.2144/btn-2018-0067 Text en © 2019 Ryan D Mohan This work is licensed under the Attribution-NonCommercial-NoDerivatives 4.0 Unported License (http://creativecommons.org/licenses/by-nc-nd/4.0/) |
spellingShingle | Benchmark Thapa, Ada Sullivan, Shea M Nguyen, Minh Q Buckley, Dominic Ngo, Vy T Dada, Austin O Blankinship, Essence Cloud, Veronica Mohan, Ryan D Brief freezing steps lead to robust immunofluorescence in the Drosophila nervous system |
title | Brief freezing steps lead to robust immunofluorescence in the Drosophila nervous system |
title_full | Brief freezing steps lead to robust immunofluorescence in the Drosophila nervous system |
title_fullStr | Brief freezing steps lead to robust immunofluorescence in the Drosophila nervous system |
title_full_unstemmed | Brief freezing steps lead to robust immunofluorescence in the Drosophila nervous system |
title_short | Brief freezing steps lead to robust immunofluorescence in the Drosophila nervous system |
title_sort | brief freezing steps lead to robust immunofluorescence in the drosophila nervous system |
topic | Benchmark |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7031821/ https://www.ncbi.nlm.nih.gov/pubmed/31687836 http://dx.doi.org/10.2144/btn-2018-0067 |
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