Live-cell nanoscopy with spontaneous blinking of conventional green fluorescent proteins

Alexey S Gavrikov; Mikhail S Baranov; Alexander S Mishin

doi:10.1016/j.bbrc.2019.11.163

Live-cell nanoscopy with spontaneous blinking of conventional green fluorescent proteins

Biochem Biophys Res Commun. 2020 Feb 19;522(4):852-854. doi: 10.1016/j.bbrc.2019.11.163. Epub 2019 Dec 1.

Authors

Alexey S Gavrikov¹, Mikhail S Baranov², Alexander S Mishin³

Affiliations

¹ Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Miklukho-Maklay St. 16/10, Moscow, 117997, Russia.
² Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Miklukho-Maklay St. 16/10, Moscow, 117997, Russia; Pirogov Russian National Research Medical University, Ostrovitianov 1, Moscow, 117997, Russia.
³ Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Miklukho-Maklay St. 16/10, Moscow, 117997, Russia. Electronic address: mishin@ibch.ru.

PMID: 31801668
DOI: 10.1016/j.bbrc.2019.11.163

Abstract

Single-molecule localization microscopy with spontaneously blinking fluorescent tags holds a promise of simplified imaging setup for live-cell application. However, robust blinking has been reported for just a few fluorescent proteins. Here we report on a comparison of spontaneous blinking for three bright green fluorescent proteins, mAvicFP1, AausFP1, and mNeonGreen. mAvicFP1 outperforms other fluorescent proteins in this list in a wide range of camera exposure times and illumination intensities. We establish imaging conditions for live-cell nanoscopy and single-particle tracking with mAvicFP1.

Keywords: Blinking; Fluorescent proteins; Live-cell nanoscopy; Single-molecule localization microscopy.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Caveolins / metabolism
Cell Survival
Green Fluorescent Proteins / metabolism*
HeLa Cells
Humans
Microscopy, Fluorescence
Nanotechnology*

Substances

Caveolins
Green Fluorescent Proteins