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A Low Percent Ethanol Method for Immobilizing Planarians

Planarians have recently become a popular model system for the study of adult stem cells, regeneration and polarity. The system is attractive for both undergraduate and graduate research labs, since planarian colonies are low cost and easy to maintain. Also in situ hybridization, immunofluorescence...

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Detalles Bibliográficos
Autores principales: Stevenson, Claire G., Beane, Wendy Scott
Formato: Texto
Lenguaje:English
Publicado: Public Library of Science 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3001876/
https://www.ncbi.nlm.nih.gov/pubmed/21179478
http://dx.doi.org/10.1371/journal.pone.0015310
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author Stevenson, Claire G.
Beane, Wendy Scott
author_facet Stevenson, Claire G.
Beane, Wendy Scott
author_sort Stevenson, Claire G.
collection PubMed
description Planarians have recently become a popular model system for the study of adult stem cells, regeneration and polarity. The system is attractive for both undergraduate and graduate research labs, since planarian colonies are low cost and easy to maintain. Also in situ hybridization, immunofluorescence and RNA-interference (RNAi) gene knockdown techniques have been developed for planarian studies. However, imaging of live worms (particularly at high magnifications) is difficult because animals are strongly photophobic; they quickly move away from light sources and out of frame. The current methods available to inhibit movement in planarians include RNAi injection and exposure to cold temperatures. The former is labor and time intensive, while the latter precludes the use of many fluorescent reporter dyes. Here, we report a simple, inexpensive and reversible method to immobilize planarians for live imaging. Our data show that a short 1 hour treatment with 3% ethanol (EtOH) is sufficient to inhibit both the fine and gross movements of Schmidtea mediterranea planarians, of the typical size used (4–6 mm), with full recovery of movement within 3–4 hours. Importantly, EtOH treatment did not interfere with regeneration, even after repeated exposure, nor lyse epithelial cells (as assayed by H&E staining). We demonstrate that a short exposure to a low concentration of EtOH is a quick and effective method of immobilizing planarians, one that is easily adaptable to planarians of all sizes and will increase the accessibility of live imaging assays to planarian researchers.
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spelling pubmed-30018762010-12-21 A Low Percent Ethanol Method for Immobilizing Planarians Stevenson, Claire G. Beane, Wendy Scott PLoS One Research Article Planarians have recently become a popular model system for the study of adult stem cells, regeneration and polarity. The system is attractive for both undergraduate and graduate research labs, since planarian colonies are low cost and easy to maintain. Also in situ hybridization, immunofluorescence and RNA-interference (RNAi) gene knockdown techniques have been developed for planarian studies. However, imaging of live worms (particularly at high magnifications) is difficult because animals are strongly photophobic; they quickly move away from light sources and out of frame. The current methods available to inhibit movement in planarians include RNAi injection and exposure to cold temperatures. The former is labor and time intensive, while the latter precludes the use of many fluorescent reporter dyes. Here, we report a simple, inexpensive and reversible method to immobilize planarians for live imaging. Our data show that a short 1 hour treatment with 3% ethanol (EtOH) is sufficient to inhibit both the fine and gross movements of Schmidtea mediterranea planarians, of the typical size used (4–6 mm), with full recovery of movement within 3–4 hours. Importantly, EtOH treatment did not interfere with regeneration, even after repeated exposure, nor lyse epithelial cells (as assayed by H&E staining). We demonstrate that a short exposure to a low concentration of EtOH is a quick and effective method of immobilizing planarians, one that is easily adaptable to planarians of all sizes and will increase the accessibility of live imaging assays to planarian researchers. Public Library of Science 2010-12-14 /pmc/articles/PMC3001876/ /pubmed/21179478 http://dx.doi.org/10.1371/journal.pone.0015310 Text en Stevenson, Beane. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Stevenson, Claire G.
Beane, Wendy Scott
A Low Percent Ethanol Method for Immobilizing Planarians
title A Low Percent Ethanol Method for Immobilizing Planarians
title_full A Low Percent Ethanol Method for Immobilizing Planarians
title_fullStr A Low Percent Ethanol Method for Immobilizing Planarians
title_full_unstemmed A Low Percent Ethanol Method for Immobilizing Planarians
title_short A Low Percent Ethanol Method for Immobilizing Planarians
title_sort low percent ethanol method for immobilizing planarians
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3001876/
https://www.ncbi.nlm.nih.gov/pubmed/21179478
http://dx.doi.org/10.1371/journal.pone.0015310
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