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Related Concept Videos

Super-resolution Fluorescence Microscopy01:37

Super-resolution Fluorescence Microscopy

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Super-resolution fluorescence microscopy (SRFM) provides a better resolution than conventional fluorescence microscopy by reducing the point spread function (PSF). PSF is the light intensity distribution from a point that causes it to appear blurred. Due to PSF, each fluorescing point appears bigger than its actual size, and it is the PSF interference of nearby fluorophores that causes the blurred image. Various approaches to achieving higher resolution through SRFM have recently been...
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Related Experiment Video

Updated: Mar 23, 2026

Whole-cell Super-Resolution Imaging via DNA-PAINT on a Spinning Disk Confocal with Optical Photon Reassignment
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Published on: January 6, 2026

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Quantitative super-resolution imaging with qPAINT.

Ralf Jungmann1,2, Maier S Avendaño1,2, Mingjie Dai1,3

  • 1Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, Massachusetts, USA.

Nature Methods
|March 29, 2016
PubMed
Summary
This summary is machine-generated.

Counting molecules precisely is difficult. Quantitative points accumulation in nanoscale topography (qPAINT) uses DNA probes for accurate molecule counting, independent of dye properties, enabling cellular applications.

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Area of Science:

  • Biophysics
  • Molecular Biology
  • Nanotechnology

Background:

  • Accurate quantification of molecules within complexes is a significant challenge in biological imaging.
  • Existing super-resolution microscopy techniques face limitations in precise molecule counting due to factors like dye photophysics.

Purpose of the Study:

  • To develop a robust and accurate method for counting molecules in biological systems.
  • To overcome the limitations of current microscopy techniques in quantitative molecular analysis.

Main Methods:

  • Utilized quantitative points accumulation in nanoscale topography (qPAINT), a method employing programmable, specific binding of dye-labeled DNA probes.
  • The qPAINT method's independence from dye photophysics ensures reliable counting across a wide dynamic range.
  • Benchmarked the method on DNA nanostructures and applied it to cellular samples.

Main Results:

  • Demonstrated high precision and accuracy in counting integer numbers of target molecules.
  • Successfully quantified proteins in situ within cellular environments.
  • Determined the number of single-molecule FISH probes bound to specific mRNA targets.

Conclusions:

  • qPAINT offers a versatile and accurate approach for quantitative molecular counting in biological research.
  • The method's robustness and wide dynamic range make it suitable for various cellular applications.
  • qPAINT advances the capability for precise molecular enumeration in complex biological systems.