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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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Long-tip high-speed atomic force microscopy for nanometer-scale imaging in live cells.

Mikihiro Shibata1, Takayuki Uchihashi2, Toshio Ando2

  • 11] Max Planck Florida Institute for Neuroscience, Jupiter, FL 33458, USA [2] Department of Neurobiology, Duke University Medical School, Durham, NC 27710, USA.

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This study introduces a novel long-tip high-speed atomic force microscopy method for imaging live cells. This technique achieves nanometer resolution, enabling visualization of cellular dynamics like filopodia and endocytosis.

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Area of Science:

  • Cell Biology
  • Biophysics
  • Microscopy

Background:

  • Visualizing live cell dynamics at nanometer resolution under physiological conditions is challenging.
  • High-speed atomic force microscopy (HS-AFM) excels at imaging biomolecules but struggles with larger samples like live mammalian cells due to tip-sample collisions.

Purpose of the Study:

  • To develop a modified HS-AFM technique capable of imaging the surface structure and dynamic morphological changes of live mammalian cells with high spatiotemporal resolution.
  • To overcome the limitations of conventional HS-AFM in imaging larger biological samples.

Main Methods:

  • Modification of HS-AFM by attaching an extremely long (~3 μm) and thin (~5 nm) amorphous carbon tip to the cantilever.
  • Imaging of live COS-7, HeLa cells, and hippocampal neurons using the modified HS-AFM system.

Main Results:

  • The long-tip HS-AFM successfully imaged the surface structure of live cells with nanometer spatial and second-level temporal resolution.
  • The technique enabled visualization of dynamic cellular processes including filopodia morphogenesis, membrane ruffles, pit formation, and endocytosis.

Conclusions:

  • Long-tip HS-AFM is a powerful tool for high-resolution, real-time imaging of live cell surface dynamics under physiological conditions.
  • This advancement expands the application of HS-AFM to complex cellular structures and processes, offering new insights into cell biology.