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Overview of Electron Microscopy01:25

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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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Immunoelectron microscopy utilizes immunogold labeling of endogenous proteins with specific antibodies to detect and localize these proteins in cells and tissues. The procedure provides insights into the distribution and quantification of protein under different stimulation conditions offering clues about their functions. Conjugating highly electron-dense gold particles with primary or secondary antibodies allow antigen detection on and within cells, with high resolution and specificity.
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The early pioneers of microscopy opened a window into the invisible world of microorganisms. In 1830, Joseph Jackson Lister created an essentially modern light microscope. The 20th century saw the development of microscopes that leveraged nonvisible light, such as fluorescence microscopy that uses an ultraviolet light source and electron microscopy that uses short-wavelength electron beams. These advances significantly improved magnification, image resolution, and contrast. By comparison, the...
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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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Some living eukaryotes during and after scanning electron microscopy.

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  • 1Department of Ecology and Environmental System, Kyungpook National University, 37224, Sangju, Republic of Korea. kiwoo@knu.ac.kr.

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|November 4, 2021
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Some organisms survive harsh scanning electron microscopy (SEM) conditions. Techniques like cryo-SEM and polymer coating enable live imaging of biological specimens, advancing microscopy capabilities.

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Area of Science:

  • Biological Sciences
  • Microscopy
  • Life Sciences

Background:

  • Electron microscopy (EM) is crucial for biological imaging.
  • Specimens often die under EM's vacuum and radiation.
  • Organisms have historically survived SEM conditions.

Purpose of the Study:

  • To explore survival of organisms under scanning electron microscopy (SEM).
  • To identify methods enabling live imaging in electron microscopy.
  • To advance preservation techniques for multicellular organisms in EM.

Main Methods:

  • Observation of surviving organisms under SEM.
  • Utilizing methods such as no chemical fixation.
  • Employing cryo-SEM and polymer coating techniques.

Main Results:

  • Bacteria, fungi, plants, and animals (beetles, ticks, tardigrades) survived SEM.
  • Specific methods (no fixation, cold stage, cryo-SEM, polymer coating) facilitated survival.
  • These techniques support preservation and extended live imaging.

Conclusions:

  • Survival of diverse organisms under SEM is documented.
  • Combined preservation techniques enhance live imaging potential.
  • Further advancements may enable long-term live imaging of multicellular organisms.