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Cathodoluminescence of green fluorescent protein exhibits the redshifted spectrum and the robustness

Green fluorescent protein (GFP) and its variants are an essential tool for visualizing functional units in biomaterials. This is achieved by the fascinating optical properties of them. Here, we report novel optical properties of enhanced GFP (EGFP), which is one of widely used engineered variants of...

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Autores principales: Akiba, Keiichirou, Tamehiro, Katsuyuki, Matsui, Koki, Ikegami, Hayata, Minoda, Hiroki
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7562900/
https://www.ncbi.nlm.nih.gov/pubmed/33060754
http://dx.doi.org/10.1038/s41598-020-74367-4
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author Akiba, Keiichirou
Tamehiro, Katsuyuki
Matsui, Koki
Ikegami, Hayata
Minoda, Hiroki
author_facet Akiba, Keiichirou
Tamehiro, Katsuyuki
Matsui, Koki
Ikegami, Hayata
Minoda, Hiroki
author_sort Akiba, Keiichirou
collection PubMed
description Green fluorescent protein (GFP) and its variants are an essential tool for visualizing functional units in biomaterials. This is achieved by the fascinating optical properties of them. Here, we report novel optical properties of enhanced GFP (EGFP), which is one of widely used engineered variants of the wild-type GFP. We study the electron-beam-induced luminescence, which is known as cathodoluminescence (CL), using the hybrid light and transmission electron microscope. Surprisingly, even from the same specimen, we observe a completely different dependences of the fluorescence and CL on the electron beam irradiation. Since light emission is normally independent of whether an electron is excited to the upper level by light or by electron beam, this difference is quite peculiar. We conclude that the electron beam irradiation causes the local generation of a new redshifted form of EGFP and CL is preferentially emitted from it. In addition, we also find that the redshifted form is rather robust to electron bombardment. These remarkable properties can be utilized for three-dimensional reconstruction without electron staining in focused ion beam/scanning electron microscopy technology and provide significant potential for simultaneously observing the functional information specified by super-resolution CL imaging and the structural information at the molecular level obtained by electron microscope.
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spelling pubmed-75629002020-10-19 Cathodoluminescence of green fluorescent protein exhibits the redshifted spectrum and the robustness Akiba, Keiichirou Tamehiro, Katsuyuki Matsui, Koki Ikegami, Hayata Minoda, Hiroki Sci Rep Article Green fluorescent protein (GFP) and its variants are an essential tool for visualizing functional units in biomaterials. This is achieved by the fascinating optical properties of them. Here, we report novel optical properties of enhanced GFP (EGFP), which is one of widely used engineered variants of the wild-type GFP. We study the electron-beam-induced luminescence, which is known as cathodoluminescence (CL), using the hybrid light and transmission electron microscope. Surprisingly, even from the same specimen, we observe a completely different dependences of the fluorescence and CL on the electron beam irradiation. Since light emission is normally independent of whether an electron is excited to the upper level by light or by electron beam, this difference is quite peculiar. We conclude that the electron beam irradiation causes the local generation of a new redshifted form of EGFP and CL is preferentially emitted from it. In addition, we also find that the redshifted form is rather robust to electron bombardment. These remarkable properties can be utilized for three-dimensional reconstruction without electron staining in focused ion beam/scanning electron microscopy technology and provide significant potential for simultaneously observing the functional information specified by super-resolution CL imaging and the structural information at the molecular level obtained by electron microscope. Nature Publishing Group UK 2020-10-15 /pmc/articles/PMC7562900/ /pubmed/33060754 http://dx.doi.org/10.1038/s41598-020-74367-4 Text en © The Author(s) 2020 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Akiba, Keiichirou
Tamehiro, Katsuyuki
Matsui, Koki
Ikegami, Hayata
Minoda, Hiroki
Cathodoluminescence of green fluorescent protein exhibits the redshifted spectrum and the robustness
title Cathodoluminescence of green fluorescent protein exhibits the redshifted spectrum and the robustness
title_full Cathodoluminescence of green fluorescent protein exhibits the redshifted spectrum and the robustness
title_fullStr Cathodoluminescence of green fluorescent protein exhibits the redshifted spectrum and the robustness
title_full_unstemmed Cathodoluminescence of green fluorescent protein exhibits the redshifted spectrum and the robustness
title_short Cathodoluminescence of green fluorescent protein exhibits the redshifted spectrum and the robustness
title_sort cathodoluminescence of green fluorescent protein exhibits the redshifted spectrum and the robustness
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7562900/
https://www.ncbi.nlm.nih.gov/pubmed/33060754
http://dx.doi.org/10.1038/s41598-020-74367-4
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