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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.
Total Internal Reflection Fluorescence Microscopy01:05

Total Internal Reflection Fluorescence Microscopy

Total internal reflection fluorescence microscopy or TIRF is an advanced microscopic technique used to visualize fluorophores in samples close to a solid surface with a higher refractive index, such as a glass coverslip. TIRF only allows fluorophores in proximity to the solid surface to be excited. When light from a medium with a lower refractive index (such as air) hits the glass coverslip at a critical angle, the light undergoes total internal reflection stead of passing through the glass.

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Updated: Jun 4, 2026

Use of Dual Optical Tweezers and Microfluidics for Single-Molecule Studies
06:53

Use of Dual Optical Tweezers and Microfluidics for Single-Molecule Studies

Published on: November 18, 2022

Ultrahigh-resolution optical trap with single-fluorophore sensitivity.

Matthew J Comstock1, Taekjip Ha, Yann R Chemla

  • 1Department of Physics and Center for the Physics of Living Cells, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA.

Nature Methods
|February 22, 2011
PubMed
Summary

We developed a novel single-molecule instrument combining optical traps and fluorescence microscopy. This tool simultaneously measures DNA mechanics and fluorescence, enabling new insights into molecular interactions.

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Combining Single-molecule Manipulation and Imaging for the Study of Protein-DNA Interactions

Published on: August 27, 2014

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Last Updated: Jun 4, 2026

Use of Dual Optical Tweezers and Microfluidics for Single-Molecule Studies
06:53

Use of Dual Optical Tweezers and Microfluidics for Single-Molecule Studies

Published on: November 18, 2022

Optical Trapping of Nanoparticles
13:39

Optical Trapping of Nanoparticles

Published on: January 15, 2013

Combining Single-molecule Manipulation and Imaging for the Study of Protein-DNA Interactions
14:43

Combining Single-molecule Manipulation and Imaging for the Study of Protein-DNA Interactions

Published on: August 27, 2014

Area of Science:

  • Biophysics
  • Molecular Biology
  • Nanotechnology

Background:

  • Understanding DNA-protein interactions is crucial for molecular biology.
  • Existing methods often lack the resolution to observe simultaneous mechanical and fluorescent changes at the single-molecule level.

Purpose of the Study:

  • To present a novel single-molecule instrument for simultaneous high-resolution mechanical and fluorescence measurements.
  • To demonstrate the instrument's capability in observing DNA oligonucleotide binding and unbinding dynamics.

Main Methods:

  • Utilizing a time-shared, ultrahigh-resolution dual optical trap interlaced with a confocal fluorescence microscope.
  • Observing single fluorophore-labeled DNA oligonucleotides binding to complementary DNA immobilized between trapped beads.
  • Simultaneously measuring angstrom-scale changes in tether extension and single-fluorophore fluorescence.

Main Results:

  • Successfully observed binding and unbinding events of DNA oligonucleotides.
  • Detected angstrom-scale mechanical changes coincident with single-fluorophore signals.
  • Determined DNA duplex melting rate as a function of applied force using fluorescence readout.

Conclusions:

  • The developed instrument enables simultaneous measurement of angstrom-scale mechanical motion and fluorescence properties of single molecules.
  • This technology opens new avenues for studying DNA-binding proteins and their dynamics at the single-molecule level.
  • Potential applications include observing DNA translocase activity and protein internal configuration changes.