Stoichiometric quantification of spatially dense assemblies with qPAINT

Baker, Matthew A. B.; Nieves, Daniel J.; Hilzenrat, Geva; Berengut, Jonathan F.; Gaus, Katharina; Lee, Lawrence K.

doi:10.1039/c9nr00472f

Cited by 13 publications

(10 citation statements)

References 23 publications

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“…Since its initial implementation in 2006, PAINT‐based SMLM has seen many innovative applications 1,2,5,7–22,36 . The development of DNA‐PAINT in 2010 represented a sea change, by providing a means to incorporate high specificity into the transient interaction.…”

Section: Discussionmentioning

confidence: 99%

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PAINT using proteins: A new brush for super‐resolution artists

Mochrie

Horrocks

et al. 2020

Protein Science

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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.

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Section: Discussionmentioning

confidence: 99%

“…In this implementation of the method, however, the transient interaction is highly specific, dictated by the sequence of the two complementary DNA strands. DNA‐PAINT has been widely used to image DNA origami type structures in vitro 9–20 …”

Section: Encoding Specificity Using Dna‐paintmentioning

confidence: 99%

PAINT using proteins: A new brush for super‐resolution artists

Mochrie

Horrocks

et al. 2020

Protein Science

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“…(e) For qPAINT calibration, a 1mer single-site nanotube is used. The images were reproduced with permission from Baker et al, published by the Royal Society of Chemistry, 2019 [53].…”

Section: Discussionmentioning

confidence: 99%

“…Moreover, qPAINT is capable of imaging the small distance site of a DNA nanotube, which would allow access to the DNA helix structure. The docking strand needs to be in a close parallel position, a gap of approximately 3 nm (Figure 12) [53]. proteins, such as antibodies and streptavidin, on nanoparticles and microparticles, with nanometric accuracy in three dimensions and multiple colors [52].…”

Section: Dna-paintmentioning

confidence: 99%

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Toward Sub-Diffraction Imaging of Single-DNA Molecule Sensors Based on Stochastic Switching Localization Microscopy

Lee

Batjikh

Kang

2020

Sensors

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The natural characteristics of deoxyribonucleic acid (DNA) enable its advanced applications in nanotechnology as a special tool that can be detected by high-resolution imaging with precise localization. Super-resolution (SR) microscopy enables the examination of nanoscale molecules beyond the diffraction limit. With the development of SR microscopy methods, DNA nanostructures can now be optically assessed. Using the specific binding of fluorophores with their target molecules, advanced single-molecule localization microscopy (SMLM) has been expanded into different fields, allowing wide-range detection at the single-molecule level. This review discusses the recent progress in the SR imaging of DNA nano-objects using SMLM techniques, such as direct stochastic optical reconstruction microscopy, binding-activated localization microscopy, and point accumulation for imaging nanoscale topography. Furthermore, we discuss their advantages and limitations, present applications, and future perspectives.

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