Peptide-PAINT Using a Transfected-Docker Enables Live- and Fixed-Cell Super-Resolution Imaging.

Autor: Maity BK; Department of Physics, University of Illinois at Urbana-Champaign, Urbana, 61801, USA., Nall D; Department of Physics, University of Illinois at Urbana-Champaign, Urbana, 61801, USA., Lee Y; Center for the Physics of the Living Cell, University of Illinois at Urbana-Champaign, Urbana, 61801, USA., Selvin PR; Department of Physics, University of Illinois at Urbana-Champaign, Urbana, 61801, USA.; Center for the Physics of the Living Cell, University of Illinois at Urbana-Champaign, Urbana, 61801, USA.; Center for Biophysics and Quantitative Biology, University of Illinois at Urbana-Champaign, Urbana, 61801, USA.
Jazyk: angličtina
Zdroj: Small methods [Small Methods] 2023 Apr; Vol. 7 (4), pp. e2201181. Date of Electronic Publication: 2023 Feb 03.
DOI: 10.1002/smtd.202201181
Abstrakt: Point accumulation for imaging in nanoscale topography (PAINT) is a single-molecule technique for super-resolution microscopy, which uses exchangeable single stranded DNA oligos or peptide-pairs to create blinking phenomenon and achieves ≈5-25 nanometer resolution. Here, it is shown that by transfecting the protein-of-interest with a docker-coil, rather than by adding the docker externally-as is the norm when using DNA tethers or antibodies as dockers-similar localization can be achieved, ≈10 nm. However, using a transfected docker has several experimental advances and simplifications. Most importantly, it allows Peptide-PAINT to be applied to transfected live cells for imaging surface proteins in mammalian cells and neurons under physiological conditions. The enhanced resolution of Peptide-PAINT is also shown for organelles in fixed cells to unravel structural details including ≈40-nm and ≈60-nm axial repeats in vimentin filaments in the cytoplasm, and fiber shapes of sub-100-nm histone-rich regions in the nucleus.
(© 2023 The Authors. Small Methods published by Wiley-VCH GmbH.)
Databáze: MEDLINE
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