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Related Concept Videos

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...
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: Jun 14, 2026

Single-Molecule Tracking Microscopy - A Tool for Determining the Diffusive States of Cytosolic Molecules
10:20

Single-Molecule Tracking Microscopy - A Tool for Determining the Diffusive States of Cytosolic Molecules

Published on: September 5, 2019

Fast, single-molecule localization that achieves theoretically minimum uncertainty.

Carlas S Smith1, Nikolai Joseph, Bernd Rieger

  • 1Department of Imaging Science and Technology, Delft University of Technology, The Netherlands.

Nature Methods
|April 6, 2010
PubMed
Summary
This summary is machine-generated.

We developed a fast algorithm for analyzing single fluorophore data, achieving optimal precision. This method enables real-time super-resolution imaging and other advanced applications.

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Last Updated: Jun 14, 2026

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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Single-Molecule Imaging of Nuclear Transport
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Single-Molecule Imaging of Nuclear Transport

Published on: June 9, 2010

Area of Science:

  • Biophysics
  • Computational Imaging
  • Data Analysis

Background:

  • Accurate parameter estimation is crucial for single-molecule localization microscopy (SMLM).
  • Existing methods may face limitations in speed and precision for real-time analysis.

Purpose of the Study:

  • To develop an iterative algorithm for precise localization and intensity estimation of single fluorophores.
  • To achieve the Cramér-Rao lower bound for parameter estimation in fluorescence imaging.

Main Methods:

  • An iterative algorithm was designed to converge to the maximum likelihood estimate.
  • The algorithm was implemented on graphics processing unit (GPU) hardware for accelerated computation.

Main Results:

  • The algorithm efficiently computes and achieves the Cramér-Rao lower bound.
  • GPU implementation processes over 10^5 fits and Cramér-Rao lower bound calculations per second.
  • This enables real-time data analysis capabilities.

Conclusions:

  • The developed algorithm provides a highly efficient and accurate method for single fluorophore analysis.
  • Real-time processing capabilities open new avenues for super-resolution imaging and other high-throughput applications.