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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.
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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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Three-Dimensional Microscopy in Microbiology

Three-dimensional imaging techniques are essential in cell biology, allowing researchers to visualize intricate cellular structures with high resolution. Two prominent methods, Differential Interference Contrast Microscopy (DIC) and Confocal Scanning Laser Microscopy (CSLM), provide distinct advantages for imaging live and thick specimens, respectively.Differential Interference Contrast MicroscopyDIC microscopy enhances contrast in transparent, unstained samples by converting phase...

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Related Experiment Video

Updated: May 17, 2026

Super-resolution Imaging of the Bacterial Division Machinery
08:47

Super-resolution Imaging of the Bacterial Division Machinery

Published on: January 21, 2013

Super-resolution imaging by localization microscopy.

Dylan M Owen1, Astrid Magenau, David J Williamson

  • 1Lowy Cancer Research Centre, Centre for Vascular Research, University of New South Wales, Sydney, Australia. Dylan.owen@unsw.edu.au

Methods in Molecular Biology (Clifton, N.J.)
|October 23, 2012
PubMed
Summary
This summary is machine-generated.

Photoactivated localization microscopy (PALM) and Stochastic optical reconstruction microscopy (STORM) offer super-resolution imaging by precisely localizing single molecules. This overview details PALM protocols for fixed cells, enabling nanoscale molecular distribution analysis.

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Last Updated: May 17, 2026

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Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells
11:06

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

Published on: June 30, 2018

Area of Science:

  • Biophysics
  • Optical Microscopy
  • Cell Biology

Background:

  • Photoactivated localization microscopy (PALM) and Stochastic optical reconstruction microscopy (STORM) are advanced super-resolution techniques.
  • These methods achieve nanoscale resolution by localizing single fluorescent molecules.
  • Commercial instruments are available, offering tens of nanometers lateral resolution.

Purpose of the Study:

  • To provide an overview of current super-resolution microscopy technologies, including live-cell and 3D PALM.
  • To present a detailed protocol for performing PALM experiments in fixed cell monolayers.
  • To enable quantitative analysis of molecular distributions at the nanoscale.

Main Methods:

  • Utilizing photoactivated localization microscopy (PALM) for super-resolution imaging.
  • Implementing a detailed protocol for instrumentation, data acquisition, and analysis.
  • Employing fixed HeLa cells expressing photo-switchable fluorescent protein PS-CFP2 targeted to the plasma membrane.

Main Results:

  • Demonstration of super-resolution imaging with tens of nanometers lateral resolution.
  • Generation of quantitative data on molecular distributions using PALM.
  • Successful application of the PALM protocol to visualize membrane-bound proteins.

Conclusions:

  • PALM is a powerful technique for nanoscale molecular imaging in fixed cells.
  • The provided protocol facilitates the acquisition and analysis of super-resolution data.
  • This methodology advances the study of molecular organization in biological systems.