Dendra2-tagged Lifeact and MAP4 as exchangeable probes for single-molecule fluorescence imaging of cytoskeleton in live cells

Elena V. Zagaynova, Olga E. Furman, Maxim M. Perfilov, Natalia V. Klementieva, Konstantin A. Lukyanov, Nina G. Bozhanova, Alexander S. Mishin

Research output: Chapter in Book/Report/Conference proceedingConference contributionpeer-review

1 Citation (Scopus)

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.

Original languageEnglish
Title of host publicationBiophotonics
Subtitle of host publicationPhotonic Solutions for Better Health Care VI
EditorsJurgen Popp, Valery V. Tuchin, Francesco Saverio Pavone
PublisherSPIE
ISBN (Electronic)9781510618961
DOIs
Publication statusPublished - 2018
Externally publishedYes
EventBiophotonics: Photonic Solutions for Better Health Care VI 2018 - Strasbourg, France
Duration: 23 Apr 201826 Apr 2018

Publication series

NameProceedings of SPIE - The International Society for Optical Engineering
Volume10685
ISSN (Print)0277-786X
ISSN (Electronic)1996-756X

Conference

ConferenceBiophotonics: Photonic Solutions for Better Health Care VI 2018
Country/TerritoryFrance
CityStrasbourg
Period23/04/1826/04/18

Keywords

  • actin
  • cytoskeleton
  • Dendra2
  • photoconversion
  • Super-resolution fluorescence microscopy
  • tubulin

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