17 May 2018 Dendra2-tagged Lifeact and MAP4 as exchangeable probes for single-molecule fluorescence imaging of cytoskeleton in live cells
Author Affiliations +
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.
© (2018) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE). Downloading of the abstract is permitted for personal use only.
Elena V. Zagaynova, Elena V. Zagaynova, Olga E. Furman, Olga E. Furman, Maxim M. Perfilov, Maxim M. Perfilov, Natalia V. Klementieva, Natalia V. Klementieva, Konstantin A. Lukyanov, Konstantin A. Lukyanov, Nina G. Bozhanova, Nina G. Bozhanova, Alexander S. Mishin, Alexander S. Mishin, } "Dendra2-tagged Lifeact and MAP4 as exchangeable probes for single-molecule fluorescence imaging of cytoskeleton in live cells", Proc. SPIE 10685, Biophotonics: Photonic Solutions for Better Health Care VI, 106850S (17 May 2018); doi: 10.1117/12.2306834; https://doi.org/10.1117/12.2306834
PROCEEDINGS
6 PAGES


SHARE
Back to Top