ID 112380
Author
Akiyama, Toshihiro University of Tsukuba
Inoko, Akihito Aichi Cancer Center Research Institute
Kaji, Yuichi University of Tsukuba
Kakiguchi, Kisa RIKEN
Segawa, Hiroki The University of Tokyo|National Research Institute of Police Science
Ishitsuka, Kei University of Tsukuba
Yoshida, Masaki University of Tsukuba
Numata, Osamu University of Tsukuba
Leproux, Philippe Institut de Recherche XLIM
Couderc, Vincent Institut de Recherche XLIM
Oshika, Tetsuro University of Tsukuba
Kano, Hideaki University of Tsukuba
Content Type
Journal Article
Description
Despite growing demand for truly naïve imaging, label-free observation of cilium-related structure remains challenging, and validation of the pertinent molecules is correspondingly difficult. In this study, in retinas and cultured cells, we distinctively visualized Rootletin filaments in rootlets in the second harmonic generation (SHG) channel, integrated in custom coherent nonlinear optical microscopy (CNOM) with a simple, compact, and ultra-broadband supercontinuum light source. This SHG signal was primarily detected on rootlets of connecting cilia in the retinal photoreceptor and was validated by colocalization with anti-Rootletin staining. Transfection of cells with Rootletin fragments revealed that the SHG signal can be ascribed to filaments assembled from the R234 domain, but not to cross-striations assembled from the R123 domain. Consistent with this, Rootletin-depleted cells lacked SHG signal expected as centrosome linker. As a proof of concept, we confirmed that similar fibrous SHG was observed even in unicellular ciliates. These findings have potential for broad applications in clinical diagnosis and biophysical experiments with various organisms.
Journal Title
Scientific Reports
ISSN
20452322
Publisher
Springer Nature
Volume
7
Start Page
39967
Published Date
2017-01-06
Remark
Supplementary Information : srep_7_39967_s1.pdf
Rights
© The Author(s) 2017
This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
EDB ID
DOI (Published Version)
URL ( Publisher's Version )
FullText File
language
eng
TextVersion
Publisher
departments
Medical Sciences