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Related Concept Videos

Overview of Electron Microscopy01:25

Overview of Electron Microscopy

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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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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.
Fundamental Principles
Accelerated...
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Transmission Electron Microscopy01:15

Transmission Electron Microscopy

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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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Immunogold Electron Microscopy01:20

Immunogold Electron Microscopy

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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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Cryo-electron Microscopy01:28

Cryo-electron Microscopy

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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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Preparation of Samples for Electron Microscopy01:20

Preparation of Samples for Electron Microscopy

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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: Feb 7, 2026

Direct Comparison of Hyperspectral Stimulated Raman Scattering and Coherent Anti-Stokes Raman Scattering Microscopy for Chemical Imaging
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Direct Comparison of Hyperspectral Stimulated Raman Scattering and Coherent Anti-Stokes Raman Scattering Microscopy for Chemical Imaging

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Electronic Preresonance Stimulated Raman Scattering Microscopy.

Lu Wei1, Wei Min1

  • 1Department of Chemistry , Columbia University , New York , New York 10027 , United States.

The Journal of Physical Chemistry Letters
|July 13, 2018
PubMed
Summary

Electronic preresonance stimulated Raman scattering (EPR-SRS) microscopy offers high sensitivity and chemical specificity, overcoming limitations of traditional optical methods. This technique enables supermultiplex imaging in biological samples by utilizing novel Raman-active dyes.

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Direct Comparison of Hyperspectral Stimulated Raman Scattering and Coherent Anti-Stokes Raman Scattering Microscopy for Chemical Imaging
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Area of Science:

  • Biophotonics and advanced optical microscopy techniques.
  • Chemical imaging and spectroscopy for biological applications.

Background:

  • Optical microscopy is crucial for research, but fluorescence microscopy lacks chemical information.
  • Vibrational imaging offers chemical specificity but often has low sensitivity or poor biocompatibility.
  • Existing methods struggle to balance sensitivity, specificity, and biocompatibility for biological imaging.

Purpose of the Study:

  • To review the utilization of electronic resonance in Raman spectroscopy and microscopy.
  • To discuss the physical principles and advantages of the electronic preresonance region.
  • To provide an outlook on the future development and applications of EPR-SRS microscopy.

Main Methods:

  • Review of prior research on electronic resonance in Raman spectroscopy.
  • Analysis of the electronic preresonance phenomenon and its enhancement factors.
  • Focus on electronic preresonance stimulated Raman scattering (EPR-SRS) microscopy.

Main Results:

  • EPR-SRS microscopy achieves high detection sensitivity and excellent vibrational specificity for chromophores.
  • Newly developed Raman-active dyes enable EPR-SRS to surpass the optical color barrier of fluorescence microscopy.
  • The method is well-suited for supermultiplex imaging in biological samples.

Conclusions:

  • EPR-SRS microscopy represents a significant advancement, combining high sensitivity with chemical specificity.
  • This technique overcomes limitations of current optical microscopy methods for biological research.
  • EPR-SRS microscopy holds broad potential for future applications in biophotonics and supermultiplex imaging.