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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 1, 2026

Conducting Multiple Imaging Modes with One Fluorescence Microscope
08:32

Conducting Multiple Imaging Modes with One Fluorescence Microscope

Published on: October 28, 2018

High-throughput single-molecule fluorescence spectroscopy using parallel detection.

X Michalet1, R A Colyer, G Scalia

  • 1Dept of Chemistry & Biochemistry, Los Angeles, CA, USA 90095.

Proceedings of Spie--The International Society for Optical Engineering
|June 1, 2011
PubMed
Summary
This summary is machine-generated.

This study introduces a multispot excitation and detection method to accelerate single-molecule fluorescence spectroscopy. This parallelization technique enhances data acquisition speed for fluorescence correlation spectroscopy (FCS) and single-molecule measurements.

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Last Updated: Jun 1, 2026

Conducting Multiple Imaging Modes with One Fluorescence Microscope
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Published on: October 28, 2018

Single-Molecule Tracking Microscopy - A Tool for Determining the Diffusive States of Cytosolic Molecules
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Published on: September 5, 2019

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High Precision FRET at Single-molecule Level for Biomolecule Structure Determination

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Area of Science:

  • Analytical Chemistry
  • Biophysics
  • Physical Chemistry

Background:

  • Single-molecule fluorescence spectroscopy offers extreme sensitivity for natural science applications.
  • Current methods require long acquisition times due to low molecule concentrations and statistical needs.

Purpose of the Study:

  • To develop a parallelized approach for faster data acquisition in single-molecule fluorescence spectroscopy.
  • To demonstrate a multispot excitation and detection geometry for improved experimental efficiency.

Main Methods:

  • Utilized a multispot excitation and detection geometry.
  • Integrated novel highly-parallel detector arrays, including multipixel single-photon counting detectors.
  • Applied the method to fluorescence correlation spectroscopy (FCS) and single-molecule fluorescence measurements.

Main Results:

  • Demonstrated a general approach for parallelizing single-molecule fluorescence spectroscopy.
  • Successfully applied the technique with novel detector arrays for faster measurements.
  • Illustrated the potential of multispot geometry for accelerating data collection.

Conclusions:

  • Parallelization using multispot geometry significantly speeds up data acquisition in single-molecule fluorescence spectroscopy.
  • The developed approach is versatile and compatible with advanced detector technologies.
  • This advancement enhances the practicality and applicability of single-molecule techniques across scientific disciplines.