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

• DOI: 10.1093/pcp/pcw027
• Publication Date: 2016-03-01T02:44:06Z
• Authors: Junko Hasegawa; Yuki Sakamoto; Satoru Nakagami; Mitsuhiro Aida; Shinichiro Sawa; Sachihiro Matsunaga
• Keywords: Clearance; Epidermis (zoology); Petal; Plant cell; Sepal; Spongy tissue; Pericycle

Clearing techniques eliminate factors that interfere with microscopic observation, including light scattering and absorption by pigments cytoplasmic components. The allow fluorescence-based detailed analyses of materials characterization the three-dimensional structure organs. We describe a simple rapid clearing imaging method, termed 'TOMEI' (Transparent plant Organ MEthod for Imaging), which enables observation intact This method involves reagent containing 2,2′-thiodiethanol. Conveniently, transparent organs were prepared within only 3–6 h. detected fluorescent stains at depth approximately 200 µm using confocal laser scanning microscopy analyzed proteins in internal tissues cleared TOMEI. adapted TOMEI various Arabidopsis thaliana Oryza sativa , leaves, flower buds, stalks, root nematode-infected root-knots. visualized whole leaves A. from adaxial epidermis to abaxial as well protoxylem metaxylem vessels vascular bundles embedded spongy mesophyll cells. Inner floral observed buds without need prepare sections or remove sepals. Multicolor dyes, based on optical are possible root-knots revealed nematodes induce giant cell expansion DNA content-dependent manner. is applicable analysis dyes quantitatively morphological characteristics