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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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Conventional electron microscopy (EM) involves dehydration, fixation, and staining of biological samples, which distorts the native state of biological molecules and results in several artifacts. Also, the high-energy electron beam damages the sample and makes it difficult to obtain high-resolution images. These issues can be addressed using cryo-EM, which uses frozen samples and gentler electron beams. The technique was developed by Jacques Dubochet, Joachim Frank, and Richard Henderson, for...
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The wavelengths of visible light ultimately limit the maximum theoretical resolution of images created by light microscopes. Most light microscopes can only magnify 1000X, and a few can magnify up to 1500X. Electrons, like electromagnetic radiation, can behave like waves, but with wavelengths of 0.005 nm, they produce significantly greater resolution up to 0.05 nm as compared to 500 nm for visible light. An electron microscope (EM) can create a sharp image that is magnified up to 2,000,000X.
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A scanning electron microscope (SEM) is used to study the surface features of a sample by using an electron beam that scans the sample surface in a two-dimensional manner. Typically, areas between ~1 centimeter to 5 micrometers in width can be imaged. SEM can be used to image bacteria, viruses, tissues as well as larger samples like insects. Conventional SEM gives a magnification ranging from 20X to 30,000X and spatial resolution of 50 to 100 nanometers.
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In 1931, physicist Ernst Ruska—building on the idea that magnetic fields can direct an electron beam just as lenses can direct a beam of light in an optical microscope—developed the first prototype of the electron microscope. This development led to the development of the field of electron microscopy. In the transmission electron microscope (TEM), electrons are produced by a hot tungsten element and accelerated by a potential difference in an electron gun, which gives them up to 400...
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Tailoring surface acoustic wave atomisation for cryo-electron microscopy sample preparation.

Dariush Ashtiani1, Alex de Marco, Adrian Neild

  • 1Department of Mechanical and Aerospace Engineering, Monash University, Clayton, Victoria, Australia. adrian.neild@monash.edu.

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|March 15, 2019
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Summary
This summary is machine-generated.

Surface acoustic wave (SAW) atomisation can be precisely controlled for cryo electron microscopy. This study optimizes SAW atomisation for minimal fluid transfer, achieving stable aerosol jets at low flow rates.

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Area of Science:

  • Physics
  • Materials Science
  • Biotechnology

Background:

  • Surface acoustic wave (SAW) atomisation is typically optimized for pharmacological delivery, focusing on droplet size and fluid volume.
  • Cryo electron microscopy (cryo-EM) grid preparation requires precise, minimal fluid transfer, necessitating a different approach to SAW atomisation characterization.

Purpose of the Study:

  • To investigate and optimize Surface Acoustic Wave (SAW) atomisation for precise, low-volume fluid deposition required in cryo-electron microscopy.
  • To analyze aerosol jet geometry (width, cone angle, elevation angle) and stability under low power conditions.

Main Methods:

  • Experiments focused on varying channel width and location for fluid delivery to the atomization site.
  • Analysis of aerosol jet characteristics, including width, cone angle, and elevation angle, at low power settings.

Main Results:

  • Achieved stable aerosol jet generation at flow rates as low as 0.55 μl s-1.
  • Demonstrated control over aerosol jet width (0.5 mm) and minimal elevation angle variation (2°).

Conclusions:

  • SAW atomisation can be adapted for high-precision, low-volume fluid transfer applications like cryo-EM grid preparation.
  • Optimizing channel geometry is key to achieving stable and controlled aerosol jets at low flow rates.