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X-ray Diffraction of Biological Samples01:10

X-ray Diffraction of Biological Samples

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X-ray diffraction or XRD is an analytical tool that utilizes X-rays to study ordered structures such as crystalline organic and inorganic samples, polycrystalline materials, proteins, carbohydrates, and drugs.
According to Bragg's law, when X-rays strike the sample positioned on a stage, the rays are  scattered by the electron clouds around the sample atoms. The  X-ray diffraction or scattering is caused by constructive interference of the X-ray waves that reflect off the internal...
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German physicist Wilhelm Röntgen (1845–1923) was experimenting with electrical current when he discovered that a mysterious and invisible "ray" would pass through his flesh but leave an outline of his bones on a screen coated with a metal compound. In 1895, Röntgen made the first durable record of the internal parts of a living human: an "X-ray" image (as it came to be called) of his wife’s hand. Scientists worldwide quickly began their own experiments with...
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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 size of the unit cell and the arrangement of atoms in a crystal may be determined from measurements of the diffraction of X-rays by the crystal, termed X-ray crystallography.
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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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Super-resolution fluorescence microscopy (SRFM) provides a better resolution than conventional fluorescence microscopy by reducing the point spread function (PSF). PSF is the light intensity distribution from a point that causes it to appear blurred. Due to PSF, each fluorescing point appears bigger than its actual size, and it is the PSF interference of nearby fluorophores that causes the blurred image. Various approaches to achieving higher resolution through SRFM have recently been...
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Updated: Apr 16, 2026

Cell Culture on Silicon Nitride Membranes and Cryopreparation for Synchrotron X-ray Fluorescence Nano-analysis
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Biostructural Science Inspired by Next-Generation X-Ray Sources.

Sol M Gruner1, Eaton E Lattman

  • 1Department of Physics.

Annual Review of Biophysics
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PubMed
Summary

Next-generation synchrotron sources enable advanced biomolecular crystallography and scattering techniques. New technologies also allow these cutting-edge methods to be used at existing facilities.

Keywords:
X-ray free-electron laserbiomolecular solution scatteringbright storage ring sourcesenergy recovery linacmicrocrystallographynanocrystallographysynchrotron radiation sources

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Area of Science:

  • Structural biology
  • Biophysics
  • Materials science

Background:

  • Advancements in synchrotron radiation sources are crucial for modern structural biology.
  • Next-generation sources offer unprecedented capabilities for probing molecular structures.

Purpose of the Study:

  • To describe novel methods in biomolecular crystallography and solution scattering.
  • To predict future trends in these research areas.
  • To highlight the dual role of next-generation sources.

Main Methods:

  • Utilizing X-ray free-electron lasers, energy recovery linacs, and ultra-low-emittance storage rings.
  • Applying serial microcrystallography techniques.
  • Implementing advanced solution scattering experiments.

Main Results:

  • Next-generation sources are driving innovation in biomolecular structure determination.
  • New technologies facilitate the use of advanced methods at existing storage rings.
  • These sources provide new capabilities and inspire method development.

Conclusions:

  • Next-generation synchrotron sources are revolutionizing structural biology.
  • Existing facilities can be upgraded to perform cutting-edge experiments.
  • The field benefits from both new source development and innovative applications at current infrastructure.