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Related Concept Videos

Atomic Force Microscopy01:08

Atomic Force Microscopy

Atomic force microscopy (AFM) is a type of scanning probe microscopy that can analyze topographic details of various specimens like ceramics, glass, polymers, and biological samples. AFM offers over 1000 times more resolution than the optical imaging system. Images generated from AFM are three-dimensional surface profiles, offering an advantage over the flat, two-dimensional images from other imaging techniques.
The AFM Probe
The probe is regarded as the heart of any AFM setup and comprises the...
Viral Structure00:56

Viral Structure

Viruses are extraordinarily diverse in shape and size, but they all have several structural features in common. All viruses have a core that contains a DNA- or RNA-based genome. The core is surrounded by a protective coat of proteins called the capsid. The capsid is composed of subunits called capsomeres. The capsid and genome-containing core are together known as the nucleocapsid.

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Sample Preparation for Single Virion Atomic Force Microscopy and Super-resolution Fluorescence Imaging
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Nanoscale structural features determined by AFM for single virus particles.

Shu-wen W Chen1, Michael Odorico, Matthieu Meillan

  • 1CEA, iBEB, Service de Biochimie et Toxicologie Nucléaire, DSV/iBEB/SBTN - Bat 170, BP17171, F-30207 Bagnols sur Cèze, France. jlpellequer@cea.fr.

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Summary

Single-image analysis of atomic force microscopy (AFM) images reveals structural details of bio-particles like tobacco mosaic virus (TMV). Resolution and processing algorithms significantly impact the scale of observed features.

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Area of Science:

  • Biophysics
  • Materials Science
  • Nanotechnology

Background:

  • Atomic Force Microscopy (AFM) is crucial for imaging bio-particles.
  • Traditional analysis often involves multi-image averaging.
  • Single-particle analysis offers a new approach for AFM data.

Purpose of the Study:

  • To introduce and validate single-image analysis for AFM data.
  • To investigate the impact of probing resolution on AFM imaging of bio-particles.
  • To demonstrate the method using tobacco mosaic virus (TMV).

Main Methods:

  • AFM imaging of TMV at different probing resolutions (1.95 and 0.49 nm/pixel).
  • Comparison with an in silico topograph derived from TMV crystal structure.
  • Application of image processing algorithms for feature extraction.

Main Results:

  • Tip convolution artifacts were observed, overestimating TMV width by ~10 nm.
  • Vertical orientation minimized tip convolution effects on height measurements.
  • Low resolution revealed helical repeats; high resolution showed individual protein subunits.

Conclusions:

  • Single-image analysis enables detailed study of bio-particle structures without limitations.
  • Probing resolution and image processing algorithms dictate the scale of discernible features in AFM.
  • This method enhances the understanding of molecular systems using AFM.