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Related Concept Videos

Super-resolution Fluorescence Microscopy01:37

Super-resolution Fluorescence Microscopy

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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...
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Protein Dynamics in Living Cells01:19

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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: Jul 27, 2025

Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells
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Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells

Published on: June 30, 2018

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Recent progress on single-molecule localization microscopy.

Lusheng Gu1,2,3, Wei Ji1,2,3

  • 1Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China.

Biophysics Reports
|June 8, 2023
PubMed
Summary
This summary is machine-generated.

Super-resolution microscopy using single-molecule localization achieves ~20 nm resolution, breaking the diffraction limit. Recent advancements enhance localization precision for studying cellular nanostructures and dynamics.

Keywords:
Centroid fittingLocalization precisionSingle-molecule localization microscopySuper-resolution imaging

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Three-dimensional Super Resolution Microscopy of F-actin Filaments by Interferometric PhotoActivated Localization Microscopy iPALM
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Related Experiment Videos

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Area of Science:

  • Biophysics
  • Optical Microscopy
  • Cell Biology

Background:

  • Single-molecule localization microscopy (SMLM) has surpassed the diffraction limit of conventional fluorescence microscopy for over a decade.
  • Initial SMLM techniques achieved resolutions around 20 nm using photoactivatable probes and centroid fitting.
  • Growing demands in biological research necessitate further improvements in localization precision.

Purpose of the Study:

  • To review recent advancements in single-molecule localization microscopy (SMLM) techniques.
  • To highlight innovations in measurement principles and hardware design for improved SMLM performance.
  • To emphasize the importance of these advanced SMLM methods for nanoscale biological investigations.

Main Methods:

  • Focus on state-of-the-art methods that have significantly improved single-molecule localization precision.
  • Discussion of novel measurement principles enabling enhanced resolution.
  • Overview of recent hardware designs contributing to SMLM performance gains.

Main Results:

  • Significant improvements in localization precision of single-molecules have been achieved.
  • New techniques have been developed that enhance the overall performance of SMLM.
  • These advancements are crucial for visualizing nanoscale structures and dynamics within cells.

Conclusions:

  • Recent developments in SMLM have substantially boosted resolution and precision.
  • Advanced SMLM is indispensable for detailed studies of cellular nanostructures.
  • These techniques are vital for understanding biomacromolecule dynamics at the nanoscale.