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Scanning Electron Microscopy01:07

Scanning Electron Microscopy

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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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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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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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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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Related Experiment Video

Updated: Jul 4, 2025

Processing Embryo, Eggshell, and Fungal Culture for Scanning Electron Microscopy
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Scanning electron microscope-based evaluation of eggshell quality.

Prem Lal Mahato1, Tina Weatherby2, Kristen Ewell2

  • 1Department of Human Nutrition Food and Animal Sciences, College of Tropical, Agriculture and Human Resources, University of Hawaii at Manoa, Honolulu, HI, USA.

Poultry Science
|February 1, 2024
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Summary

This study developed a new method using scanning electron microscopy (SEM) and energy-dispersive spectrometry (EDS) to precisely measure eggshell thickness and mineral content. This technique aids in selecting breeding stock for improved eggshell quality.

Keywords:
eggshell-qualitymammillarymineralpalisadescanning electron microscope

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Area of Science:

  • Materials Science
  • Animal Science
  • Biochemistry

Background:

  • Eggshell quality is vital for the poultry industry, impacting economic viability.
  • Poor eggshell quality leads to significant economic losses.
  • Accurate methods are needed to assess eggshell quality for breeding improvements.

Purpose of the Study:

  • To develop a precise method for determining eggshell thickness using SEM.
  • To analyze eggshell mineral components with SEM-Energy Dispersive Spectrometry (EDS).
  • To establish a scientific basis for selecting superior breeding stock for enhanced eggshells.

Main Methods:

  • Utilized Scanning Electron Microscopy (SEM) to visualize eggshell structure.
  • Employed SEM-Energy Dispersive Spectrometry (EDS) for mineral component analysis.
  • Analyzed four types of table eggs (Cage-Free Organic, Caged Non-Organic from US Mainland and Hawaii).

Main Results:

  • Identified three distinct eggshell layers: cuticle, palisade, and mammillary regions.
  • Cage-Free Organic eggs from Hawaii exhibited greater eggshell thickness.
  • Observed variations in mineral distribution, with calcium being the most abundant.

Conclusions:

  • The developed SEM and EDS method accurately assesses eggshell quality and biochemical properties.
  • This methodology serves as a valuable tool for selecting breeding stock with superior eggshell traits.
  • Improved eggshell quality can be achieved in future generations through targeted breeding programs.