Author:
Akiba Keiichirou,Tamehiro Katsuyuki,Matsui Koki,Ikegami Hayata,Minoda Hiroki
Abstract
AbstractGreen 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.
Funder
Ministry of Education, Culture, Sports, Science and Technology
Publisher
Springer Science and Business Media LLC
Reference23 articles.
1. Chalfie, M., Tu, Y., Euskirchen, G., Ward, W. W. & Prasher, D. C. Green fluorescent protein as a marker for gene expression. Science 263, 802–805 (1994).
2. Tsien, R. Y. The green fluorescence protein. Annu. Rev. Biochem. 67, 509–544 (1998).
3. Jung, G. Fluorescent Protein I: From Understanding to Design (Springer, New York, 2013).
4. Egerton, R. F., Li, P. & Malac, M. Radiation damage in the TEM and SEM. Micron 35, 399–409 (2004).
5. Fisher, P. J., Wessels, W. S., Dietz, A. B. & Prendergast, F. G. Enhanced biological cathodoluminescence. Opt. Commun. 281, 1901–1908 (2008).
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