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Rapid clearing of biological organs by using phosphoric acid, a hydrophilic solution with high refractive index

View ORCID ProfileMasakazu Umezawa, Shinsuke Haruguchi, Rihito Fukushima, Shota Sekiyama, Masao Kamimura, Kohei Soga
doi: https://doi.org/10.1101/452425
Masakazu Umezawa
1Department of Materials Science and Technology, Faculty of Industrial Science and Technology, Tokyo University of Science, Katsushika-ku, Tokyo, Japan
2Imaging Frontier Center, Research Institute for Science and Technology, Tokyo University of Science, Noda-shi, Chiba, Japan
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  • ORCID record for Masakazu Umezawa
  • For correspondence: masa-ume@rs.noda.tus.ac.jp mail@ksoga.com
Shinsuke Haruguchi
1Department of Materials Science and Technology, Faculty of Industrial Science and Technology, Tokyo University of Science, Katsushika-ku, Tokyo, Japan
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Rihito Fukushima
1Department of Materials Science and Technology, Faculty of Industrial Science and Technology, Tokyo University of Science, Katsushika-ku, Tokyo, Japan
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Shota Sekiyama
1Department of Materials Science and Technology, Faculty of Industrial Science and Technology, Tokyo University of Science, Katsushika-ku, Tokyo, Japan
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Masao Kamimura
1Department of Materials Science and Technology, Faculty of Industrial Science and Technology, Tokyo University of Science, Katsushika-ku, Tokyo, Japan
2Imaging Frontier Center, Research Institute for Science and Technology, Tokyo University of Science, Noda-shi, Chiba, Japan
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Kohei Soga
1Department of Materials Science and Technology, Faculty of Industrial Science and Technology, Tokyo University of Science, Katsushika-ku, Tokyo, Japan
2Imaging Frontier Center, Research Institute for Science and Technology, Tokyo University of Science, Noda-shi, Chiba, Japan
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  • For correspondence: masa-ume@rs.noda.tus.ac.jp mail@ksoga.com
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Abstract

Tissue clearing is a fundamental challenge in biology and medicine to achieve high-resolution optical imaging of tissues deep inside intact organs. The clearing methods, reported up to now, require long incubation time or physical/electrical pressure to achieve tissue clearing, which is done by matching the refractive indices of the whole sample and medium to that of the lipid layer. Here we show that phosphoric acid increases the refractive index of the medium and can increase the transparency of formalin-fixed tissue samples rapidly. Immersion of fixed tissues of mice in phosphoric acid solutions increased their transparency within 60 min in the case of 3-mm-thick fixed tissue specimens. While phosphoric acid suppresses bright signals on the boundary of cells in their phase-contrast images, it does not damage the morphology of cell membrane with phospholipid bilayer. The protocol presented herein may contribute to develop better and faster soaking methods for tissue clearing than previously reported protocols.

Highlights

  • ▪ Phosphoric acid can reduce light scattering by tissue samples.

  • ▪ Tissue clearing effect of phosphoric acid is fast and needs only 60-min incubation.

  • ▪ Cell membrane was preserved during incubation using phosphoric acid.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.
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Posted October 24, 2018.
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Rapid clearing of biological organs by using phosphoric acid, a hydrophilic solution with high refractive index
Masakazu Umezawa, Shinsuke Haruguchi, Rihito Fukushima, Shota Sekiyama, Masao Kamimura, Kohei Soga
bioRxiv 452425; doi: https://doi.org/10.1101/452425
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Rapid clearing of biological organs by using phosphoric acid, a hydrophilic solution with high refractive index
Masakazu Umezawa, Shinsuke Haruguchi, Rihito Fukushima, Shota Sekiyama, Masao Kamimura, Kohei Soga
bioRxiv 452425; doi: https://doi.org/10.1101/452425

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