Abstract
PAINT (points accumulation for imaging in nanoscale topography) refers to methods that achieve the sparse temporal labeling required for super‐resolution imaging by using transient interactions between a biomolecule of interest and a fluorophore. There have been a variety of different implementations of this method since it was first described in 2006. Recent papers illustrate how transient peptide–protein interactions, rather than small molecule binding or DNA oligonucleotide duplex formation, can be employed to perform PAINT‐based single molecule localization microscopy (SMLM). We discuss the different approaches to PAINT using peptide and protein interactions, and their applications in vitro and in vivo. We highlight the important parameters to consider when selecting suitable peptide–protein interaction pairs for such studies. We also note the opportunities for protein scientists to apply their expertise in guiding the choice of peptide and protein pairs that are used. Finally, we discuss the potential for expanding super‐resolution imaging methods based on transient peptide–protein interactions, including the development of simultaneous multicolor imaging of multiple proteins and the study of very high and very low abundance proteins in live cells.
Highlights
Recent papers describe the successful use of transient peptide–protein interactions to perform super-resolution microscopy,[1,2,3] in particular as a new way to implement the method known as points accumulation for imaging in nanoscale topography (PAINT)[4] (Figure 1)
These studies demonstrate that different implementations of PAINT, employing protein–peptide interaction pairs, enable high resolution single-molecule localization microscopy (SMLM) in vitro, in fixed cells, and inside live cells
Recent work, which is the focus of this review, has demonstrated the potential for using peptide–protein interactions, rather than DNA duplex formation, to achieve transient labeling
Summary
Recent papers describe the successful use of transient peptide–protein interactions to perform super-resolution microscopy,[1,2,3] in particular as a new way to implement the method known as points accumulation for imaging in nanoscale topography (PAINT)[4] (Figure 1).These studies demonstrate that different implementations of PAINT, employing protein–peptide interaction pairs, enable high resolution single-molecule localization microscopy (SMLM) in vitro, in fixed cells, and inside live cells. KEYWORDS coiled coil, fluorescence microscopy, live cell imaging, peptide, protein, super-resolution, TPR Recent papers describe the successful use of transient peptide–protein interactions to perform super-resolution microscopy,[1,2,3] in particular as a new way to implement the method known as points accumulation for imaging in nanoscale topography (PAINT)[4] (Figure 1).
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