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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: May 8, 2026

Imaging Intermediate Filaments and Microtubules with 2-dimensional Direct Stochastic Optical Reconstruction Microscopy
14:23

Imaging Intermediate Filaments and Microtubules with 2-dimensional Direct Stochastic Optical Reconstruction Microscopy

Published on: March 6, 2018

Single-Molecule Localization Microscopy with Fixed Photoactivatable Fluorescent Proteins and Direct Stochastic

Rumelo Amor1, Anusha Malapaka2, Alex J McCann2

  • 1Queensland Brain Institute, The University of Queensland, Brisbane, QLD, Australia.

Methods in Molecular Biology (Clifton, N.J.)
|May 6, 2026
PubMed
Summary
This summary is machine-generated.

Single-molecule localization microscopy (SMLM) techniques like Photoactivated Localization Microscopy (PALM) and Stochastic Optical Reconstruction Microscopy (STORM) offer super-resolution imaging. Protocols are provided for fixed-cell PALM and direct Stochastic Optical Reconstruction Microscopy (dSTORM) data acquisition and analysis.

Keywords:
Fixed PALMSMLMdSTORM

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Last Updated: May 8, 2026

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Published on: March 6, 2018

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Simultaneous Multicolor Imaging of Biological Structures with Fluorescence Photoactivation Localization Microscopy
12:51

Simultaneous Multicolor Imaging of Biological Structures with Fluorescence Photoactivation Localization Microscopy

Published on: December 9, 2013

Area of Science:

  • Cell Biology
  • Microscopy
  • Biophysics

Background:

  • Fluorescence microscopy is crucial for visualizing cellular structures and protein localization.
  • Single-molecule localization microscopy (SMLM) surpasses the diffraction limit for enhanced spatial resolution.
  • Techniques like PALM and STORM provide sub-diffraction limit access to subcellular organization.

Purpose of the Study:

  • Introduce the principles of SMLM.
  • Position PALM and (d)STORM within super-resolution microscopy.
  • Provide detailed protocols for fixed-cell PALM and dSTORM imaging.

Main Methods:

  • Utilizes fixed Photoactivated Localization Microscopy (PALM) and direct Stochastic Optical Reconstruction Microscopy (dSTORM).
  • Involves temporal separation of fluorophore emission events for precise molecular localization.
  • Employs Zeiss ZEN Black 2012 and Abbelight NEO software for image reconstruction and analysis.

Main Results:

  • Fixed PALM and (d)STORM remain widely used due to molecular specificity and broad applicability.
  • These methods enable visualization of subcellular organization below the diffraction limit.
  • Detailed protocols facilitate image acquisition and data analysis for specific cell types.

Conclusions:

  • SMLM techniques, particularly fixed PALM and (d)STORM, are powerful tools for cell biology research.
  • The provided protocols are compatible with standard SMLM platforms.
  • These methods offer broad applicability across diverse biological systems for detailed cellular analysis.