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Super-Resolution Imaging to Study Co-Localization of Proteins and Synaptic Markers in Primary Neurons

Super-Resolution Imaging to Study Co-Localization of Proteins and Synaptic Markers in Primary Neurons

Published on: October 31, 2020

PAINT using proteins: A new brush for super-resolution artists.

Curran Oi^1,2, Simon G J Mochrie^2,3, Mathew H Horrocks⁴

¹Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, Connecticut, USA.

Protein Science : a Publication of the Protein Society

|September 19, 2020

Summary

Transient peptide-protein interactions enable advanced super-resolution imaging through PAINT (points accumulation for imaging in nanoscale topography). This method allows for precise visualization of biomolecules, expanding possibilities in live-cell studies.

Keywords:

TPR coiled coil fluorescence microscopy live cell imaging peptide protein super-resolution

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Area of Science:

Biophysics
Microscopy
Molecular Biology

Background:

Points Accumulation for Imaging in Nanoscale Topography (PAINT) is a super-resolution imaging technique utilizing transient biomolecule-fluorophore interactions.
Various PAINT implementations exist, with recent advancements focusing on transient peptide-protein interactions.

Purpose of the Study:

To review and discuss PAINT methods employing peptide-protein interactions for single-molecule localization microscopy (SMLM).
To highlight applications, key selection parameters for interaction pairs, and future potential of this approach.

Main Methods:

Review of existing literature on PAINT techniques.
Focus on transient peptide-protein interactions as the labeling mechanism.
Discussion of in vitro and in vivo applications.

Main Results:

Peptide-protein interactions offer a viable alternative to small molecules or DNA for PAINT-based SMLM.
Identification of critical parameters for selecting effective peptide-protein pairs.
Demonstration of in vitro and in vivo utility.

Conclusions:

Transient peptide-protein interactions represent a powerful tool for super-resolution imaging.
Expertise in protein science is valuable for optimizing PAINT strategies.
Future directions include multicolor imaging and studying proteins of varying abundance in live cells.