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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

Photoactivated Localization Microscopy with Bimolecular Fluorescence Complementation (BiFC-PALM)
12:42

Photoactivated Localization Microscopy with Bimolecular Fluorescence Complementation (BiFC-PALM)

Published on: December 22, 2015

Single Particle Tracking Photo-Activated Localization Microscopy.

Alex J McCann1, Christopher Small1, Frédéric A Meunier2,3

  • 1Clem Jones Centre for Ageing Dementia Research (CJCADR), Queensland 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.

Super-resolution imaging techniques like single particle tracking Photo-Activated Localization Microscopy (sptPALM) allow real-time tracking of synaptic proteins. This advances our understanding of synaptic plasticity and neurotransmission at the nanoscale.

Keywords:
EndocytosisExocytosisFluorophoreNanoclusteringNeurotransmissionPhotoconversionPlasma membraneSingle-particle trackingSynapseTIRF microscopy

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Last Updated: May 8, 2026

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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:

  • Neuroscience
  • Cell Biology
  • Biophysics

Background:

  • Synaptic plasticity involves dynamic nanoscale reorganization of synaptic components.
  • Confocal microscopy has resolution limits for visualizing these rapid, small-scale changes.
  • Super-resolution microscopy, particularly SMLM, offers nanoscale resolution for real-time synaptic studies.

Purpose of the Study:

  • To provide a conceptual overview of single particle tracking Photo-Activated Localization Microscopy (sptPALM).
  • To detail the principles, implementation, and impact of sptPALM in neurotransmission research.
  • To present a protocol for sptPALM imaging of synaptic proteins in neuronal cultures.

Main Methods:

  • Utilizes total internal reflection fluorescence (TIRF) microscopy.
  • Employs sparse, stochastic activation of fluorophores for single-molecule tracking.
  • Applies single-molecule localization microscopy (SMLM) for nanoscale resolution.

Main Results:

  • Enables real-time tracking of individual synaptic proteins.
  • Allows determination of protein mobility and clustering patterns.
  • Provides nanoscale localization precision for synaptic protein behavior.

Conclusions:

  • sptPALM is a powerful technique for unraveling molecular mechanisms of synaptic transmission and plasticity.
  • The described protocol facilitates detailed analysis of synaptic protein dynamics.
  • This approach enhances understanding of nanoscale synaptic organization and function.