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Protocol for immunofluorescence staining of murine helminth-infected intestinal and lung tissues
Preserving structurally intact tissue through proper tissue fixation and cryopreservation minimizes tissue autolysis, desiccation, and enzymatic degradation. In this protocol, we describe the use of an optimized fresh freezing technique that cryopreserves the tissue and favors the retention of the n...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10322902/ https://www.ncbi.nlm.nih.gov/pubmed/37086410 http://dx.doi.org/10.1016/j.xpro.2023.102208 |
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author | El-Naccache, Darine W. Chen, Fei Gause, William C. |
author_facet | El-Naccache, Darine W. Chen, Fei Gause, William C. |
author_sort | El-Naccache, Darine W. |
collection | PubMed |
description | Preserving structurally intact tissue through proper tissue fixation and cryopreservation minimizes tissue autolysis, desiccation, and enzymatic degradation. In this protocol, we describe the use of an optimized fresh freezing technique that cryopreserves the tissue and favors the retention of the natural protein structure of antigens. We also detail the use of cold, low-concentration paraformaldehyde (PFA) fixation to enhance tissue morphology and detection of fluorescent proteins, such as GFP and TdTomato, in tissues of genetically engineered mice. For complete details on the use and execution of this protocol, please refer to Chen et al. (2022)(1) and El-Naccache et al. (2022).(2) |
format | Online Article Text |
id | pubmed-10322902 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-103229022023-07-07 Protocol for immunofluorescence staining of murine helminth-infected intestinal and lung tissues El-Naccache, Darine W. Chen, Fei Gause, William C. STAR Protoc Protocol Preserving structurally intact tissue through proper tissue fixation and cryopreservation minimizes tissue autolysis, desiccation, and enzymatic degradation. In this protocol, we describe the use of an optimized fresh freezing technique that cryopreserves the tissue and favors the retention of the natural protein structure of antigens. We also detail the use of cold, low-concentration paraformaldehyde (PFA) fixation to enhance tissue morphology and detection of fluorescent proteins, such as GFP and TdTomato, in tissues of genetically engineered mice. For complete details on the use and execution of this protocol, please refer to Chen et al. (2022)(1) and El-Naccache et al. (2022).(2) Elsevier 2023-04-21 /pmc/articles/PMC10322902/ /pubmed/37086410 http://dx.doi.org/10.1016/j.xpro.2023.102208 Text en © 2023. 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 El-Naccache, Darine W. Chen, Fei Gause, William C. Protocol for immunofluorescence staining of murine helminth-infected intestinal and lung tissues |
title | Protocol for immunofluorescence staining of murine helminth-infected intestinal and lung tissues |
title_full | Protocol for immunofluorescence staining of murine helminth-infected intestinal and lung tissues |
title_fullStr | Protocol for immunofluorescence staining of murine helminth-infected intestinal and lung tissues |
title_full_unstemmed | Protocol for immunofluorescence staining of murine helminth-infected intestinal and lung tissues |
title_short | Protocol for immunofluorescence staining of murine helminth-infected intestinal and lung tissues |
title_sort | protocol for immunofluorescence staining of murine helminth-infected intestinal and lung tissues |
topic | Protocol |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10322902/ https://www.ncbi.nlm.nih.gov/pubmed/37086410 http://dx.doi.org/10.1016/j.xpro.2023.102208 |
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