Jump to main content
Jump to site search


Protein labeling for live cell fluorescence microscopy with a highly photostable renewable signal

Abstract

We present Protein-PAINT – the implementation of general principles of PAINT (Point Accumulation for Imaging in Nanoscale Topography) for live-cell protein labeling. Our method employs specific binding of cell-permeable fluorogenic dyes to genetically encoded protein tags. We engineered three mutants of bacterial lipocalin Blc that possess different affinities to a fluorogenic dye and exhibit strong increase in fluorescence intensity upon binding. This allows for rapid labeling and washout of intracellular targets in a time scale from seconds to a few minutes. We demonstrate an order of magnitude higher photostability of fluorescence signal in comparison with spectrally similar fluorescent proteins. Protein-PAINT ensures prolonged super-resolution fluorescence microscopy of living cells in both single molecule detection and stimulated emission depletion regimes.

Back to tab navigation

Supplementary files

Publication details

The article was received on 11 Apr 2017, accepted on 01 Aug 2017 and first published on 03 Aug 2017


Article type: Edge Article
DOI: 10.1039/C7SC01628J
Citation: Chem. Sci., 2017, Accepted Manuscript
  • Open access: Creative Commons BY license
  •   Request permissions

    Protein labeling for live cell fluorescence microscopy with a highly photostable renewable signal

    N. G. Bozhanova, M. S. Baranov, N. V. Klementieva, K. S. Sarkisyan, A. S. Gavrikov, I. V. Yampolsky, E. V. Zagaynova, S. Lukyanov, K. Lukyanov and A. S. Mishin, Chem. Sci., 2017, Accepted Manuscript , DOI: 10.1039/C7SC01628J

    This article is licensed under a Creative Commons Attribution 3.0 Unported Licence. Material from this article can be used in other publications provided that the correct acknowledgement is given with the reproduced material.

    Reproduced material should be attributed as follows:

    • For reproduction of material from NJC:
      [Original citation] - Published by The Royal Society of Chemistry (RSC) on behalf of the Centre National de la Recherche Scientifique (CNRS) and the RSC.
    • For reproduction of material from PCCP:
      [Original citation] - Published by the PCCP Owner Societies.
    • For reproduction of material from PPS:
      [Original citation] - Published by The Royal Society of Chemistry (RSC) on behalf of the European Society for Photobiology, the European Photochemistry Association, and RSC.
    • For reproduction of material from all other RSC journals:
      [Original citation] - Published by The Royal Society of Chemistry.

    Information about reproducing material from RSC articles with different licences is available on our Permission Requests page.

Search articles by author

Spotlight

Advertisements