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Nat Methods. 2016 May;13(5):439-42. doi: 10.1038/nmeth.3804. Epub 2016 Mar 28.

Quantitative super-resolution imaging with qPAINT.

Author information

1
Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, Massachusetts, USA.
2
Department of Systems Biology, Harvard Medical School, Boston, Massachusetts, USA.
3
Harvard Program in Biophysics, Harvard University, Boston, Massachusetts, USA.
4
Rosenstiel Basic Medical Sciences Research Center, Department of Biology, Brandeis University, Waltham, Massachusetts, USA.

Abstract

Counting molecules in complexes is challenging, even with super-resolution microscopy. Here, we use the programmable and specific binding of dye-labeled DNA probes to count integer numbers of targets. This method, called quantitative points accumulation in nanoscale topography (qPAINT), works independently of dye photophysics for robust counting with high precision and accuracy over a wide dynamic range. qPAINT was benchmarked on DNA nanostructures and demonstrated for cellular applications by quantifying proteins in situ and the number of single-molecule FISH probes bound to an mRNA target.

PMID:
27018580
PMCID:
PMC4941813
DOI:
10.1038/nmeth.3804
[Indexed for MEDLINE]
Free PMC Article

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