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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.
Confocal Fluorescence Microscopy01:16

Confocal Fluorescence Microscopy

Confocal microscopy is an advanced microscopic technique. The prime advantage of the confocal microscope over other microscopy techniques is its ability to block the out-of-focus light from the illuminated samples using pinholes. It is widely used with fluorescence optics to obtain high-resolution, sharp contrast images. Unlike optical microscopes, confocal microscopes use a focused beam of light laser to scan the entire sample surface at different z-planes. These microscopes are, therefore,...

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Related Experiment Video

Updated: May 9, 2026

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

A user's guide to localization-based super-resolution fluorescence imaging.

Graham T Dempsey1

  • 1Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts, USA.

Methods in Cell Biology
|August 13, 2013
PubMed
Summary
This summary is machine-generated.

Stochastic Optical Reconstruction Microscopy (STORM) offers super-resolution imaging for biological samples. This guide details choosing probes, preparing samples, and acquiring/analyzing data for nanoscale biological investigations.

Keywords:
FPALMFluorescenceLocalizationPALMPhotoswitchingProbesSTORMSuper-resolution

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Last Updated: May 9, 2026

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Area of Science:

  • Biophysics
  • Optical Microscopy
  • Nanotechnology

Background:

  • Super-resolution microscopy techniques have advanced far-field fluorescence imaging.
  • Stochastic Optical Reconstruction Microscopy (STORM) achieves nanoscale resolution by localizing individual fluorophores sequentially.
  • STORM offers a conceptually and practically accessible method for biological investigations.

Purpose of the Study:

  • To provide a comprehensive guide for applying STORM to biological samples.
  • To detail critical considerations for successful STORM implementation.
  • To facilitate high-resolution nanoscale imaging in biological research.

Main Methods:

  • Discussion of photoswitchable fluorescent probe selection.
  • Guidance on biological sample preparation for STORM.
  • Considerations for selecting hardware for data acquisition.
  • Methods for data collection and analysis for image reconstruction.

Main Results:

  • The chapter provides a framework for applying STORM.
  • It addresses key experimental parameters for optimal results.
  • It enables researchers to overcome practical challenges in STORM implementation.

Conclusions:

  • STORM is a powerful technique for nanoscale biological imaging.
  • Careful consideration of probes, sample prep, hardware, and data analysis is crucial.
  • This guide empowers researchers to effectively utilize STORM for biological discovery.