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Proceedings Paper

Dendra2-tagged Lifeact and MAP4 as exchangeable probes for single-molecule fluorescence imaging of cytoskeleton in live cells
Author(s): Elena V. Zagaynova M.D.; Olga E. Furman; Maxim M. Perfilov; Natalia V. Klementieva; Konstantin A. Lukyanov; Nina G. Bozhanova; Alexander S. Mishin
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Paper Abstract

Recently, a new method called image reconstruction by integrating exchangeable single-molecule localization (IRIS) was developed (Kiuchi et al., Nat. Methods 2015, 12, 743-746). IRIS ensures high-density labeling for super-resolution microscopy but can be applied for fixed cells only. Here, we extended the IRIS conception to live cell imaging. We applied Lifeact and MAP4 transiently binding to microfilaments and microtubules, respectively. Green-to-red photoconvertible fluorescent protein Dendra2 was chosen as an efficient tag for single-molecule localization microscopy. Live-cell single-molecule localization imaging with Dendra2-Lifeact and Dendra2-MAP4 was performed. As a result, super-resolved images of actin and tubulin in dynamics in living cells were reconstructed. Importantly, Dendra2-Lifeact provided a higher number of individual localizations and denser labeling of microfilaments in comparison with commonly used Dendra2-actin.

Paper Details

Date Published: 17 May 2018
PDF: 6 pages
Proc. SPIE 10685, Biophotonics: Photonic Solutions for Better Health Care VI, 106850S (17 May 2018); doi: 10.1117/12.2306834
Show Author Affiliations
Elena V. Zagaynova M.D., Privolzhsky Research Medical Univ. (Russian Federation)
Olga E. Furman, Privolzhsky Research Medical Univ. (Russian Federation)
Maxim M. Perfilov, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry (Russian Federation)
Natalia V. Klementieva, Privolzhsky Research Medical Univ. (Russian Federation)
Konstantin A. Lukyanov, Privolzhsky Research Medical Univ. (Russian Federation)
Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry (Russian Federation)
Nina G. Bozhanova, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry (Russian Federation)
Alexander S. Mishin, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry (Russian Federation)


Published in SPIE Proceedings Vol. 10685:
Biophotonics: Photonic Solutions for Better Health Care VI
Jürgen Popp; Valery V. Tuchin; Francesco Saverio Pavone, Editor(s)

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