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Related Concept Videos

Super-resolution Fluorescence Microscopy01:37

Super-resolution Fluorescence Microscopy

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 developed.

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Related Experiment Video

Updated: May 11, 2026

Three-dimensional Super Resolution Microscopy of F-actin Filaments by Interferometric PhotoActivated Localization Microscopy (iPALM)
11:57

Three-dimensional Super Resolution Microscopy of F-actin Filaments by Interferometric PhotoActivated Localization Microscopy (iPALM)

Published on: December 1, 2016

Video-rate nanoscopy using sCMOS camera-specific single-molecule localization algorithms.

Fang Huang1, Tobias M P Hartwich, Felix E Rivera-Molina

  • 1Department of Cell Biology, Yale University School of Medicine, New Haven, Connecticut, USA.

Nature Methods
|May 28, 2013
PubMed
Summary
This summary is machine-generated.

New algorithms overcome readout noise in scientific complementary metal-oxide semiconductor (sCMOS) cameras, enabling precise single-molecule localization and super-resolution imaging up to 32 images per second in cells.

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Single Molecule Fluorescence Microscopy on Planar Supported Bilayers

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Related Experiment Videos

Last Updated: May 11, 2026

Three-dimensional Super Resolution Microscopy of F-actin Filaments by Interferometric PhotoActivated Localization Microscopy (iPALM)
11:57

Three-dimensional Super Resolution Microscopy of F-actin Filaments by Interferometric PhotoActivated Localization Microscopy (iPALM)

Published on: December 1, 2016

Video-rate Scanning Confocal Microscopy and Microendoscopy
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Video-rate Scanning Confocal Microscopy and Microendoscopy

Published on: October 20, 2011

Single Molecule Fluorescence Microscopy on Planar Supported Bilayers
20:00

Single Molecule Fluorescence Microscopy on Planar Supported Bilayers

Published on: October 31, 2015

Area of Science:

  • Biophysics
  • Optical Microscopy
  • Super-resolution Imaging

Background:

  • Scientific complementary metal-oxide semiconductor (sCMOS) cameras offer enhanced speed and field of view for nanoscopy.
  • Pixel-dependent readout noise in sCMOS cameras limits localization precision and introduces artifacts in single-molecule switching nanoscopy.

Purpose of the Study:

  • To develop algorithms that mitigate sCMOS readout noise for improved single-molecule localization.
  • To achieve unbiased and precise single-molecule localization at the theoretical limit.

Main Methods:

  • Development of novel algorithms to correct for pixel-dependent readout noise.
  • Integration of these algorithms with a multi-emitter fitting approach.
  • Application in single-molecule switching nanoscopy for super-resolution imaging.

Main Results:

  • Algorithms successfully overcome sCMOS readout noise limitations.
  • Achieved unbiased and precise single-molecule localization.
  • Demonstrated super-resolution imaging at rates up to 32 reconstructed images per second.

Conclusions:

  • The developed algorithms significantly enhance the performance of sCMOS cameras for single-molecule localization super-resolution microscopy.
  • Enables high-speed imaging in both fixed and living cells, advancing biological research.