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Three-Dimensional Imaging of Plant Organs Using a Simple and Rapid Transparency Technique

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dc.contributor.author Hasegawa, Junko en
dc.contributor.author Sakamoto, Yuki en
dc.contributor.author Nakagami, Satoru en
dc.contributor.author Aida, Mitsuhiro en
dc.contributor.author Sawa, Shinichiro en
dc.contributor.author Matsunaga, Sachihiro en
dc.date.accessioned 2016-03-11T00:57:46Z en
dc.date.available 2017-03-01 en
dc.date.issued 2016-02-29 en
dc.identifier.issn 1471-9053 en
dc.identifier.uri http://hdl.handle.net/10061/10474 en
dc.description.abstract Clearing techniques eliminate factors that interfere with microscopic observation, including light scattering and absorption by pigments and cytoplasmic components. The techniques allow fluorescence-based detailed analyses of materials and characterization of the three-dimensional structure of organs. We describe a simple and rapid clearing and imaging method, termed ‘TOMEI’ (Transparent plant Organ MEthod for Imaging), which enables microscopic observation of intact plant organs. This method involves a clearing reagent containing 2,2′-thiodiethanol. Conveniently, transparent plant organs were prepared within only 3-6 h. We detected fluorescent stains at a depth of approximately 200 µm using confocal laser scanning microscopy and analyzed fluorescent proteins in internal tissues of transparent organs cleared using TOMEI. We adapted TOMEI for various plant organs of Arabidopsis thaliana and Oryza sativa, including leaves, flower buds, flower stalks, root and nematode-infected root-knots. We visualized whole leaves of A. thaliana from the adaxial epidermis to the abaxial epidermis as well as protoxylem and metaxylem vessels of vascular bundles embedded in spongy mesophyll cells. Inner floral organs were observed in flower buds cleared using TOMEI without the need to prepare sections or remove sepals. Multicolor imaging of fluorescent proteins and dyes, and analyses of the three-dimensional structure of plant organs based on optical sections are possible using TOMEI. We analyzed root-knots cleared using TOMEI and revealed that nematodes induce giant cell expansion in a DNA content-dependent manner. The TOMEI method is applicable to analysis of fluorescent proteins and dyes quantitatively with cell morphological characteristics in whole plant organs. ja
dc.language.iso en en
dc.publisher Oxford University Press en
dc.relation.isreplacedby http://pcp.oxfordjournals.org/content/early/2016/02/29/pcp.pcw027 en
dc.rights 出版社許諾条件により、本文は2017年3月1日以降公開 The publisher require to delay the availability of full text until 2017/3/1. ja
dc.rights This is a pre-copyedited, author-produced PDF of an article accepted for publication in Plant & Cell Phsyology following peer review. The version of record Plant Cell Physiol First published online: February 29, 2016 is available online at: http://pcp.oxfordjournals.org/content/early/2016/02/29/pcp.pcw027. en
dc.subject Clearing method en
dc.subject Deep imaging en
dc.subject Fluorescent protein en
dc.subject 2,2′-Thiodiethanol ja
dc.subject Transparency technique en
dc.title Three-Dimensional Imaging of Plant Organs Using a Simple and Rapid Transparency Technique en
dc.type.nii Journal Article en
dc.textversion author en
dc.textversion publisher en
dc.identifier.jtitle Plant & Cell Physiology en
dc.relation.doi 10.1093/pcp/pcw027 en
dc.identifier.NAIST-ID 73294100 en

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