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X-ray Diffraction of Biological Samples01:10

X-ray Diffraction of Biological Samples

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 crystal...
Cryo-electron Microscopy01:28

Cryo-electron Microscopy

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...
Three-Dimensional Microscopy in Microbiology01:28

Three-Dimensional Microscopy in Microbiology

Three-dimensional imaging techniques are essential in cell biology, allowing researchers to visualize intricate cellular structures with high resolution. Two prominent methods, Differential Interference Contrast Microscopy (DIC) and Confocal Scanning Laser Microscopy (CSLM), provide distinct advantages for imaging live and thick specimens, respectively.Differential Interference Contrast MicroscopyDIC microscopy enhances contrast in transparent, unstained samples by converting phase...
Phase Contrast and Differential Interference Contrast Microscopy01:26

Phase Contrast and Differential Interference Contrast Microscopy

Phase-Contrast Microscopes
In-phase-contrast microscopes, interference between light directly passing through a cell and light refracted by cellular components is used to create high-contrast, high-resolution images without staining. It is the oldest and simplest type of microscope that creates an image by altering the wavelengths of light rays passing through the specimen. Altered wavelength paths are created using an annular stop in the condenser. The annular stop produces a hollow cone of...
Super-resolution Fluorescence Microscopy01:37

Super-resolution Fluorescence Microscopy

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 developed.
Determination of Crystal Structures01:29

Determination of Crystal Structures

In the late 1800s, the revelation that light extended beyond visible wavelengths led to the discovery of X-rays by Wilhelm Roentgen. Recognized as high-energy electromagnetic radiation with short wavelengths, X-rays prompted exploration into their interaction with crystals. Max von Laue proposed in 1912 that the periodic arrangement of atoms, ions, or molecules in crystals would cause them to diffract X-rays, a hypothesis confirmed through experiments with copper sulfate and zinc sulfide...

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Related Experiment Video

Updated: May 13, 2026

A 3D Cartographic Description of the Cell by Cryo Soft X-ray Tomography
08:47

A 3D Cartographic Description of the Cell by Cryo Soft X-ray Tomography

Published on: March 15, 2021

Imaging fully hydrated whole cells by coherent x-ray diffraction microscopy.

Daewoong Nam1, Jaehyun Park, Marcus Gallagher-Jones

  • 1RIKEN SPring-8 Center, 1-1-1 Kouto, Sayo, Hyogo 679-5148, Japan.

Physical Review Letters
|March 19, 2013
PubMed
Summary

This study introduces wet coherent x-ray diffraction microscopy for nanoscale imaging of hydrated, unstained biological specimens. The technique clearly visualizes whole cell morphologies and internal structures with resolution better than 25 nm.

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Last Updated: May 13, 2026

A 3D Cartographic Description of the Cell by Cryo Soft X-ray Tomography
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Published on: March 15, 2021

Microcrystallography of Protein Crystals and In Cellulo Diffraction
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Area of Science:

  • Biophysics
  • Microscopy
  • Cell Biology

Background:

  • High-resolution nanoscale imaging of native biological specimens is challenging.
  • Existing methods often require staining or specific sample preparation, altering native conditions.
  • Direct visualization of internal structures in intact, hydrated cells remains a significant hurdle.

Purpose of the Study:

  • To develop a novel microscopy technique for imaging fully hydrated and unstained biological specimens.
  • To achieve nanoscale resolution of internal cellular structures without contrast loss.
  • To overcome limitations of current nanoscale imaging methods for native biological samples.

Main Methods:

  • Introduction of wet coherent x-ray diffraction microscopy.
  • Utilizing hydrated, unstained biological specimens.
  • Applying coherent x-ray diffraction principles in a wet environment.

Main Results:

  • Demonstrated capability for imaging fully hydrated and unstained biological specimens.
  • Achieved clear visualization of whole cell morphologies.
  • Resolved internal cellular structures with resolution better than 25 nanometers.
  • Maintained image quality without contrast degradation.

Conclusions:

  • Wet coherent x-ray diffraction microscopy enables high-resolution nanoscale imaging of native biological samples.
  • The technique offers a new approach for studying cellular structures in their physiological state.
  • This advancement has significant implications for cell biology and biophysics research.