RPD

Research Promotion Division (RPD),

Institute of Transformative Bio-Molecules (ITbM), Nagoya University

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Molecule Gallery:

#11

Overcoming the obstacles to fluorescence microscopy in plants

The discovery of fluorescent proteins has allowed for rapid progress in the analysis of the functions and structures of organic tissues through observation of the behavior of proteins. An important part of this process is making the structure to be observed transparent so that the target protein can be seen.

However, plant cells are a more difficult target for fluorescence microscopy than animal cells, as they contain autofluorescent substances such as chlorophyll, and observation is further complicated by the refractive index of other components such as the cell wall and surrounding air.

Using chemical compound screening, researchers at ITbM have discovered a highly elutive surfactant that removes autofluorescence in plants while causing very little tissue damage, and a second compound that not only renders the observed tissue transparent but also preserves the fluorescent proteins. When a plant specimen was immersed in a solution of these two compounds, its tissue was rendered transparent in just a few days.

A truly innovative and easy to use reagent

Previously, it was necessary to thinly slice plant tissues in order to observe their internal structures at high resolution. The individual images would then have to be reconstructed into a 3D image in a lengthy and complex process.

ClearSee changes this in one fell swoop. Now, anybody can quickly and easily render plant tissue transparent and observe its internal structure without the need for special microscopy techniques. Since its discovery, the researchers have further developed it to be applicable to a wider range of plants and tissues, naming this improved version ClearSeeAlpha. ClearSeeAlpha is expected to become a valuable tool for the acceleration of plant science research.

Reference:

  • “ClearSee: a rapid optical clearing reagent for whole-plant fluorescence imaging” by Daisuke Kurihara, Yoko Mizuta, Yoshikatsu Sato, Tetsuya Higashiyama, Development 2015, 142 (23): 4168–4179. DOI: 10.1242/dev.127613
  • “ClearSeeAlpha: Advanced Optical Clearing for Whole-Plant Imaging” by Daisuke Kurihara, Yoko Mizuta, Shiori Nagahara, Tetsuya Higashiyama, Plant and Cell Physiology 2021, 62 (8): 1302–1310. DOI: 10.1093/pcp/pcab033

 

CONTACT

Research Promotion Division (RPD),

Institute of Transformative Bio-Molecules (ITbM), Nagoya University

 

530, ITbM, Furo-cho, Chikusa-ku, Nagoya 464-8601 JAPAN

E-mail: rpd[at]itbm.nagoya-u.ac.jp

 

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