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To be visualized by an electron microscope, either transmission or scanning, biological samples need to be fixed (stabilized) so the electron beam does not destroy them and dried thoroughly (desiccated/dehydrated) so the vacuum does not affect them. Fixation needs to be done as quickly as possible because the sample properties will start changing as soon as it is removed from its natural environment. For example, in a tissue sample, the oxygen levels begin decreasing, causing an altered...
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Graphene specimen support technique for low voltage STEM imaging.

Masao Yamashita1, Matthew Ryan Leyden1, Hidehito Adaniya1

  • 1Okinawa Institute of Science and Technology Graduate University, 1919-1 Tancha, Onna-son, Kunigami-gun, Okinawa 904-0495, Japan.

Microscopy (Oxford, England)
|April 29, 2017
PubMed
Summary
This summary is machine-generated.

Low voltage scanning transmission electron microscopy (STEM) using graphene supports improves imaging of viruses like bacteriophage T4. A spin sedimentation method aids sample loading onto hydrophobic graphene grids for enhanced biological sample visualization.

Keywords:
FE-SEMRaman spectroscopySTEMbacteriophagegraphenelow energy

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Area of Science:

  • Materials Science
  • Microscopy
  • Structural Biology

Background:

  • Scanning transmission electron microscopy (STEM) offers sub-nanometer resolution for imaging nanometer-sized objects.
  • Graphene, a single-atom thick conductive material, is a promising support for low voltage STEM.
  • Imaging delicate biological specimens like viruses requires advanced microscopy techniques.

Purpose of the Study:

  • To evaluate graphene films as support substrates for low voltage STEM imaging of bacteriophage T4.
  • To demonstrate the enhancement of image signal and fine structure visualization using graphene supports.
  • To address challenges in applying hydrophilic biological samples to hydrophobic graphene grids.

Main Methods:

  • Low voltage STEM imaging of bacteriophage T4.
  • Utilizing highly cleaned graphene films as specimen support.
  • Employing methylamine vanadate stain for biological samples.
  • Implementing a spin sedimentation technique for sample loading.

Main Results:

  • Graphene support films significantly improve image signal in low voltage STEM.
  • Clear visualization of bacteriophage T4 fine structure, particularly the tail, was achieved.
  • The combination of graphene support and methylamine vanadate stain enhanced low voltage STEM imaging.
  • Spin sedimentation effectively facilitated the loading of hydrophilic samples onto hydrophobic graphene grids.

Conclusions:

  • Ultrathin graphene films are highly effective specimen supports for low voltage STEM, enhancing image quality for biological samples.
  • The developed method, combining graphene support, specific staining, and spin sedimentation, enables detailed visualization of viral structures.
  • This approach offers a valuable tool for structural biology research using advanced electron microscopy techniques.