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Algae to angiosperms: Autofluorescence for rapid visualization of plant anatomy among diverse taxa
PREMISE: Fluorescence microscopy is an effective tool for viewing plant internal anatomy. However, using fluorescent antibodies or labels hinders throughput. We present a minimal protocol that takes advantage of inherent autofluorescence and aldehyde‐induced fluorescence in plant cellular and subcel...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8272585/ https://www.ncbi.nlm.nih.gov/pubmed/34268017 http://dx.doi.org/10.1002/aps3.11437 |
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author | Pegg, Timothy J. Gladish, Daniel K. Baker, Robert L. |
author_facet | Pegg, Timothy J. Gladish, Daniel K. Baker, Robert L. |
author_sort | Pegg, Timothy J. |
collection | PubMed |
description | PREMISE: Fluorescence microscopy is an effective tool for viewing plant internal anatomy. However, using fluorescent antibodies or labels hinders throughput. We present a minimal protocol that takes advantage of inherent autofluorescence and aldehyde‐induced fluorescence in plant cellular and subcellular structures to markedly increase throughput in cellular and ultrastructural visualization. METHODS AND RESULTS: Twelve species distributed across the plant phylogeny were each subjected to five fixative treatments: 1% paraformaldehyde and 2% glutaraldehyde, 2% paraformaldehyde, 2% glutaraldehyde, formalin‐acid‐alcohol (FAA), and 70% ethanol. Samples were prepared by embedding and mechanically sectioning or via whole mount. A confocal laser scanning system was used to collect micrographs. We evaluated and compared fixative influence on sample structural preservation and tissue autofluorescence. CONCLUSIONS: Formaldehyde fixation of Viridiplantae taxa samples generates useful structural data while requiring no additional histological staining or clearing. In addition, a fluorescence‐capable microscope is the only specialized equipment required for image acquisition. The minimal protocol developed in this experiment enables high‐throughput sample processing by eliminating the need for multi‐day preparations. |
format | Online Article Text |
id | pubmed-8272585 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-82725852021-07-14 Algae to angiosperms: Autofluorescence for rapid visualization of plant anatomy among diverse taxa Pegg, Timothy J. Gladish, Daniel K. Baker, Robert L. Appl Plant Sci Protocol Note PREMISE: Fluorescence microscopy is an effective tool for viewing plant internal anatomy. However, using fluorescent antibodies or labels hinders throughput. We present a minimal protocol that takes advantage of inherent autofluorescence and aldehyde‐induced fluorescence in plant cellular and subcellular structures to markedly increase throughput in cellular and ultrastructural visualization. METHODS AND RESULTS: Twelve species distributed across the plant phylogeny were each subjected to five fixative treatments: 1% paraformaldehyde and 2% glutaraldehyde, 2% paraformaldehyde, 2% glutaraldehyde, formalin‐acid‐alcohol (FAA), and 70% ethanol. Samples were prepared by embedding and mechanically sectioning or via whole mount. A confocal laser scanning system was used to collect micrographs. We evaluated and compared fixative influence on sample structural preservation and tissue autofluorescence. CONCLUSIONS: Formaldehyde fixation of Viridiplantae taxa samples generates useful structural data while requiring no additional histological staining or clearing. In addition, a fluorescence‐capable microscope is the only specialized equipment required for image acquisition. The minimal protocol developed in this experiment enables high‐throughput sample processing by eliminating the need for multi‐day preparations. John Wiley and Sons Inc. 2021-07-02 /pmc/articles/PMC8272585/ /pubmed/34268017 http://dx.doi.org/10.1002/aps3.11437 Text en © 2021 The Authors. Applications in Plant Sciences published by Wiley Periodicals LLC on behalf of Botanical Society of America https://creativecommons.org/licenses/by-nc/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc/4.0/ (https://creativecommons.org/licenses/by-nc/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes. |
spellingShingle | Protocol Note Pegg, Timothy J. Gladish, Daniel K. Baker, Robert L. Algae to angiosperms: Autofluorescence for rapid visualization of plant anatomy among diverse taxa |
title | Algae to angiosperms: Autofluorescence for rapid visualization of plant anatomy among diverse taxa |
title_full | Algae to angiosperms: Autofluorescence for rapid visualization of plant anatomy among diverse taxa |
title_fullStr | Algae to angiosperms: Autofluorescence for rapid visualization of plant anatomy among diverse taxa |
title_full_unstemmed | Algae to angiosperms: Autofluorescence for rapid visualization of plant anatomy among diverse taxa |
title_short | Algae to angiosperms: Autofluorescence for rapid visualization of plant anatomy among diverse taxa |
title_sort | algae to angiosperms: autofluorescence for rapid visualization of plant anatomy among diverse taxa |
topic | Protocol Note |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8272585/ https://www.ncbi.nlm.nih.gov/pubmed/34268017 http://dx.doi.org/10.1002/aps3.11437 |
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