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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.
Protein Dynamics in Living Cells01:19

Protein Dynamics in Living Cells

Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...

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Related Experiment Video

Updated: Jun 5, 2026

Automated System for Single Molecule Fluorescence Measurements of Surface-immobilized Biomolecules
10:57

Automated System for Single Molecule Fluorescence Measurements of Surface-immobilized Biomolecules

Published on: November 2, 2009

Single-molecule fluorescence characterization in native environment.

Thomas P Burghardt1, Katalin Ajtai

  • 1Department of Biochemistry and Molecular Biology, Mayo Clinic Rochester, Rochester, MN 55905, USA.

Biophysical Reviews
|December 24, 2010
PubMed
Summary
This summary is machine-generated.

Single-molecule detection (SMD) using fluorescence microscopy offers detailed biomolecule insights, overcoming ensemble limitations. Advanced probes enable precise imaging in dense cellular environments, revealing native molecular behavior.

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Conducting Multiple Imaging Modes with One Fluorescence Microscope
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Conducting Multiple Imaging Modes with One Fluorescence Microscope

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Last Updated: Jun 5, 2026

Automated System for Single Molecule Fluorescence Measurements of Surface-immobilized Biomolecules
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Published on: November 2, 2009

Time-Resolved Fluorescence Anisotropy from Single Molecules for Characterizing Local Flexibility in Biomolecules
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Published on: April 25, 2025

Conducting Multiple Imaging Modes with One Fluorescence Microscope
08:32

Conducting Multiple Imaging Modes with One Fluorescence Microscope

Published on: October 28, 2018

Area of Science:

  • Biophysics
  • Microscopy
  • Molecular Biology

Background:

  • Single-molecule detection (SMD) with fluorescence is a key technique for characterizing biomolecule structure and function.
  • Modern light microscopy, utilizing high numerical aperture objectives and sensitive CCD cameras, enables imaging of fluorescently tagged biomolecules.
  • Single-molecule imaging provides bottom-up characterization, avoiding limitations inherent in ensemble measurements.

Purpose of the Study:

  • To highlight the capabilities of modern fluorescence microscopy for single-molecule detection.
  • To discuss the development of specialized probes for SMD applications.
  • To explore the significance of SMD in native, high-density cellular environments.

Main Methods:

  • Utilizing advanced light microscopy with high numerical aperture objectives and sensitive CCD cameras.
  • Employing fluorescent protein and organic tags for biomolecule labeling.
  • Developing and applying specialized probes for high-density SMD.

Main Results:

  • Achieved single-particle localization precision of approximately 1 nm, circumventing the diffraction limit.
  • Enabled detection of both probe position and orientation in cellular environments.
  • Demonstrated the feasibility of SMD under conditions mimicking the native biomolecule environment.

Conclusions:

  • Single-molecule imaging offers unambiguous biomolecule characterization.
  • Specialized probes enhance SMD capabilities in dense cellular regions.
  • Native, high-density SMD is crucial for understanding molecular crowding effects on biomolecule behavior.