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タイトル: | Improved chemical fixation of lipid-secreting plant cells for transmission electron microscopy |
著者: | Kiyoto, Shingo Ichino, Takuji Awano, Tatsuya https://orcid.org/0000-0002-4735-4506 (unconfirmed) Yazaki, Kazufumi |
著者名の別形: | 清都, 晋吾 市野, 琢爾 粟野, 達也 矢﨑, 一史 |
キーワード: | imidazole Lithospermum erythrorhizon malachite green p-phenylenediamine shikonin transmission electron microscopy |
発行日: | Aug-2022 |
出版者: | Oxford University Press (OUP) |
誌名: | Microscopy |
巻: | 71 |
号: | 4 |
開始ページ: | 206 |
終了ページ: | 213 |
抄録: | Cultured Lithospermum erythrorhizon cells were fixed with a new fixation method to visualize the metabolism of shikonin derivatives, the lipophilic naphthoquinone pigments in Boraginaceae. The new fixation method combined glutaraldehyde containing malachite green, imidazole–osmium and p-phenylenediamine treatments, and cells were then observed with a transmission electron microscope. The method prevented the extraction of lipids, including shikonin derivatives, and improved the visualization of subcellular structures, especially the membrane system, when compared with that of conventional fixation. The improved quality of the transmission electron micrographs is because malachite green ionically binds to the plasma membrane, organelles and lipids and acts as a mordant for electron staining with osmium tetroxide. Imidazole promotes the reaction of osmium tetroxide, leading to enhanced electron staining. p-Phenylenediamine reduces osmium tetroxide bound to cellular materials and increases the electron density. This protocol requires only three additional reagents over conventional chemical fixation using glutaraldehyde and osmium tetroxide. |
著作権等: | © The Author(s) 2022. Published by Oxford University Press on behalf of The Japanese Society of Microscopy. This is an Open Access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited. |
URI: | http://hdl.handle.net/2433/275698 |
DOI(出版社版): | 10.1093/jmicro/dfac018 |
PubMed ID: | 35388424 |
出現コレクション: | 学術雑誌掲載論文等 |
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