Transmission electron microscopy methodology applied to the study of seedling roots
Autor(a) principal: | |
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Data de Publicação: | 2019 |
Outros Autores: | , |
Tipo de documento: | Artigo |
Idioma: | por |
Título da fonte: | Revista Brasileira Multidisciplinar |
Texto Completo: | http://revistarebram.com/index.php/revistauniara/article/view/505 |
Resumo: | The use of the transmission electron microscope (MET) is very widespread in the study of biological materials, since it allows the definition of intracellular images, allowing studies of cellular morphology and general aspects of organelles. The images are produced by the initial interaction of an electron beam with the sample, which is maintained under high vacuum conditions. Both the vacuum environment and the irradiation of the electrons during observation, represent adverse conditions for the sample, especially in the case of organic or biological materials, which are by nature hydrated, fragile and electrically insulating. Considering that there are several preparation techniques for MET, but few detailed description of these techniques in the literature, especially for the preparation of seedling roots, this work aimed to apply and adapt these techniques for the study of good quality of seedling roots. The samples were first washed in phosphate buffered saline solution, passed through the primary fixation with glutaraldehyde, then postfixed with osmium tetroxide, where they were subsequently dehydrated with acetone and infiltrated with Epon resin, finishing with polymerization, ultramicrotomy and sample staining. However, this detailed description of the methodology applied to seedlings root can be used safely and effectively by other people, since there are almost no adapted descriptions of MET methodology in the literature, especially for seedlings. |
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Transmission electron microscopy methodology applied to the study of seedling rootsMetodologia de microscopia eletrônica de transmissão aplicada ao estudo de raízes de plântulasTransmission Electron MicroscopeAdapting techniquesWeed roots.Microscópio Eletrônico de TransmissãoAdaptação de técnicasraízes de planta daninha.The use of the transmission electron microscope (MET) is very widespread in the study of biological materials, since it allows the definition of intracellular images, allowing studies of cellular morphology and general aspects of organelles. The images are produced by the initial interaction of an electron beam with the sample, which is maintained under high vacuum conditions. Both the vacuum environment and the irradiation of the electrons during observation, represent adverse conditions for the sample, especially in the case of organic or biological materials, which are by nature hydrated, fragile and electrically insulating. Considering that there are several preparation techniques for MET, but few detailed description of these techniques in the literature, especially for the preparation of seedling roots, this work aimed to apply and adapt these techniques for the study of good quality of seedling roots. The samples were first washed in phosphate buffered saline solution, passed through the primary fixation with glutaraldehyde, then postfixed with osmium tetroxide, where they were subsequently dehydrated with acetone and infiltrated with Epon resin, finishing with polymerization, ultramicrotomy and sample staining. However, this detailed description of the methodology applied to seedlings root can be used safely and effectively by other people, since there are almost no adapted descriptions of MET methodology in the literature, especially for seedlings. O emprego do microscópio eletrônico de transmissão (MET) é bastante difundido no estudo de materiais biológicos, pois ele permite definição de imagens intracelulares, permitindo estudos de morfologia celular e de aspectos gerais das organelas. As imagens são produzidas por meio da interação inicial de um feixe de elétrons com a amostra, que é mantida sob condições de alto vácuo. Tanto o ambiente de vácuo como a própria irradiação dos elétrons durante a observação, representam condições adversar para a amostra, especialmente no caso de materiais orgânicos ou biológicos, que são por natureza hidratados, frágeis e elétricamente isolantes. Considerando que existem várias técnicas de preparo para MET, mas pouca descrição detalhada destas técnicas na literatura, principalmente para o preparo de raízes de plântulas, assim, esse trabalho teve como objetivo aplicar e adaptar essas técnicas para o estudo de boa qualidade de raízes de plântulas. As amostras primeiramente foram lavadas em tampão fosfato salina, passando pela fixação primária com glutaraldeído, depois pós-fixação com tetróxido de Ósmio, onde posteriormente foram desidratadas com acetona e infiltradas com resina Epon, finalizando com a polimerização, ultramicrotomia e contrastação da amostra. Contudo, essa descrição detalhada da metodologia aplicada para raiz de plântulas, pode ser utilizada com segurança e eficácia por outras pessoas, pois quase não existem descrições adaptadas de metodologia de MET na literatura, principalmente para raiz de plântulas.Revista Brasileira Multidisciplinar - ReBraM2019-01-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfhttp://revistarebram.com/index.php/revistauniara/article/view/50510.25061/2527-2675/ReBraM/2019.v22i1.505Revista Brasileira Multidisciplinar - ReBraM; v. 22 n. 1 (2019): Janeiro-Abril; 175-1862527-26751415-3580reponame:Revista Brasileira Multidisciplinarinstname:Universidade de Araraquara (UNIARA)instacron:UNIARAporhttp://revistarebram.com/index.php/revistauniara/article/view/505/pdfCopyright (c) 2019 Revista Brasileira Multidisciplinarinfo:eu-repo/semantics/openAccessPergo, Érica MarusaBritta, Elizandra AparecidaGalletti, Patrícia Aparecida2019-05-03T20:57:18Zoai:ojs.revistarebram.com:article/505Revistahttp://revistarebram.com/index.php/revistauniaraPRIhttps://revistarebram.com/index.php/revistauniara/oairevistauniara@uniara.com||2527-26752527-2675opendoar:2019-05-03T20:57:18Revista Brasileira Multidisciplinar - Universidade de Araraquara (UNIARA)false |
dc.title.none.fl_str_mv |
Transmission electron microscopy methodology applied to the study of seedling roots Metodologia de microscopia eletrônica de transmissão aplicada ao estudo de raízes de plântulas |
title |
Transmission electron microscopy methodology applied to the study of seedling roots |
spellingShingle |
Transmission electron microscopy methodology applied to the study of seedling roots Pergo, Érica Marusa Transmission Electron Microscope Adapting techniques Weed roots. Microscópio Eletrônico de Transmissão Adaptação de técnicas raízes de planta daninha. |
title_short |
Transmission electron microscopy methodology applied to the study of seedling roots |
title_full |
Transmission electron microscopy methodology applied to the study of seedling roots |
title_fullStr |
Transmission electron microscopy methodology applied to the study of seedling roots |
title_full_unstemmed |
Transmission electron microscopy methodology applied to the study of seedling roots |
title_sort |
Transmission electron microscopy methodology applied to the study of seedling roots |
author |
Pergo, Érica Marusa |
author_facet |
Pergo, Érica Marusa Britta, Elizandra Aparecida Galletti, Patrícia Aparecida |
author_role |
author |
author2 |
Britta, Elizandra Aparecida Galletti, Patrícia Aparecida |
author2_role |
author author |
dc.contributor.author.fl_str_mv |
Pergo, Érica Marusa Britta, Elizandra Aparecida Galletti, Patrícia Aparecida |
dc.subject.por.fl_str_mv |
Transmission Electron Microscope Adapting techniques Weed roots. Microscópio Eletrônico de Transmissão Adaptação de técnicas raízes de planta daninha. |
topic |
Transmission Electron Microscope Adapting techniques Weed roots. Microscópio Eletrônico de Transmissão Adaptação de técnicas raízes de planta daninha. |
description |
The use of the transmission electron microscope (MET) is very widespread in the study of biological materials, since it allows the definition of intracellular images, allowing studies of cellular morphology and general aspects of organelles. The images are produced by the initial interaction of an electron beam with the sample, which is maintained under high vacuum conditions. Both the vacuum environment and the irradiation of the electrons during observation, represent adverse conditions for the sample, especially in the case of organic or biological materials, which are by nature hydrated, fragile and electrically insulating. Considering that there are several preparation techniques for MET, but few detailed description of these techniques in the literature, especially for the preparation of seedling roots, this work aimed to apply and adapt these techniques for the study of good quality of seedling roots. The samples were first washed in phosphate buffered saline solution, passed through the primary fixation with glutaraldehyde, then postfixed with osmium tetroxide, where they were subsequently dehydrated with acetone and infiltrated with Epon resin, finishing with polymerization, ultramicrotomy and sample staining. However, this detailed description of the methodology applied to seedlings root can be used safely and effectively by other people, since there are almost no adapted descriptions of MET methodology in the literature, especially for seedlings. |
publishDate |
2019 |
dc.date.none.fl_str_mv |
2019-01-01 |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://revistarebram.com/index.php/revistauniara/article/view/505 10.25061/2527-2675/ReBraM/2019.v22i1.505 |
url |
http://revistarebram.com/index.php/revistauniara/article/view/505 |
identifier_str_mv |
10.25061/2527-2675/ReBraM/2019.v22i1.505 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.relation.none.fl_str_mv |
http://revistarebram.com/index.php/revistauniara/article/view/505/pdf |
dc.rights.driver.fl_str_mv |
Copyright (c) 2019 Revista Brasileira Multidisciplinar info:eu-repo/semantics/openAccess |
rights_invalid_str_mv |
Copyright (c) 2019 Revista Brasileira Multidisciplinar |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Revista Brasileira Multidisciplinar - ReBraM |
publisher.none.fl_str_mv |
Revista Brasileira Multidisciplinar - ReBraM |
dc.source.none.fl_str_mv |
Revista Brasileira Multidisciplinar - ReBraM; v. 22 n. 1 (2019): Janeiro-Abril; 175-186 2527-2675 1415-3580 reponame:Revista Brasileira Multidisciplinar instname:Universidade de Araraquara (UNIARA) instacron:UNIARA |
instname_str |
Universidade de Araraquara (UNIARA) |
instacron_str |
UNIARA |
institution |
UNIARA |
reponame_str |
Revista Brasileira Multidisciplinar |
collection |
Revista Brasileira Multidisciplinar |
repository.name.fl_str_mv |
Revista Brasileira Multidisciplinar - Universidade de Araraquara (UNIARA) |
repository.mail.fl_str_mv |
revistauniara@uniara.com|| |
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1797174606303330304 |