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Related Concept Videos

Atomic Force Microscopy01:08

Atomic Force Microscopy

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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...
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Time-Lapse Single-Biomolecule Atomic Force Microscopy Investigation on Modified Graphite in Solution.

Evgeniy V Dubrovin1,2,3, Marc Schächtele1, Dmitry V Klinov2

  • 1University of Tübingen , Institute of Applied Physics, Auf der Morgenstelle 10, 72076 Tübingen, Germany.

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This study explores modified graphite as an alternative substrate for atomic force microscopy (AFM) of biomolecules. Modified graphite offers controlled ionic strength, complementing mica for single-molecule biophysics.

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Area of Science:

  • Biophysics
  • Surface Science
  • Biochemistry

Background:

  • Atomic force microscopy (AFM) is crucial for single-molecule biophysics.
  • Mica is a common AFM substrate but its high surface charge can alter biomolecular processes.
  • Investigating alternative substrates is needed to control ionic strength near the surface.

Purpose of the Study:

  • To evaluate highly oriented pyrolytic graphite (HOPG) modified with stearylamine and oligoglycine-hydrocarbon derivative (GM) monolayers as an AFM substrate.
  • To assess the behavior of adsorbed biomolecules, including plasmid DNA and E. coli RNA polymerase σ70 subunit holoenzyme (RNAP), on these modified HOPG surfaces in solution.
  • To explore the potential for real-time AFM studies of biomolecular processes like transcription on modified graphite.

Main Methods:

  • Utilized atomic force microscopy (AFM) in solution.
  • Employed modified highly oriented pyrolytic graphite (HOPG) surfaces, including those with stearylamine and GM monolayers.
  • Studied the adsorption and behavior of plasmid DNA and E. coli RNA polymerase σ70 subunit holoenzyme (RNAP).

Main Results:

  • Demonstrated ionic-strength-dependent DNA mobility on GM-modified HOPG.
  • Observed nativelike dimensions for RNAP molecules adsorbed on modified HOPG surfaces.
  • Proposed a method for real-time AFM investigation of transcription on stearylamine-modified graphite.

Conclusions:

  • Modified graphite surfaces provide a controllable ionic strength environment for AFM studies.
  • These modified graphite substrates can serve as a valuable complement to mica for investigating biomolecules and biomolecular processes.
  • This approach enables more accurate studies of biopolymer adsorption and protein-DNA interactions by mitigating substrate-induced ionic strength effects.