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Related Concept Videos

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...
Imaging Biological Samples with Optical Microscopy01:18

Imaging Biological Samples with Optical Microscopy

Optical microscopy uses optic principles to provide detailed images of samples. Antonie van Leeuwenhoek designed the first compound optical microscope in the 17th century to visualize blood cells, bacteria, and yeast cells. In 1830, Joseph Jackson Lister created an essentially modern light microscope. The 20th century saw the development of microscopes with enhanced magnification and resolution.
In optical microscopy, the specimen to be viewed is placed on a glass slide and clipped on the stage...
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.

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Related Experiment Video

Updated: Jun 22, 2026

Low-Cost Cryo-Light Microscopy Stage Fabrication for Correlated Light/Electron Microscopy
10:00

Low-Cost Cryo-Light Microscopy Stage Fabrication for Correlated Light/Electron Microscopy

Published on: June 5, 2011

High-aperture cryogenic light microscopy.

M A Le Gros1, G McDermott, M Uchida

  • 1Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, California, USA.

Journal of Microscopy
|July 2, 2009
PubMed
Summary
This summary is machine-generated.

We developed new cryogenic microscopy methods to improve fluorescence probe lifetimes and imaging resolution. This technique allows detailed observation of live cells before cryo-fixation and high-resolution analysis.

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Last Updated: Jun 22, 2026

Low-Cost Cryo-Light Microscopy Stage Fabrication for Correlated Light/Electron Microscopy
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Workflow Using a Cryogenic Coincident Fluorescence, Electron, and Ion Beam Microscope for Targeted Milling of Cells
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Workflow Using a Cryogenic Coincident Fluorescence, Electron, and Ion Beam Microscope for Targeted Milling of Cells

Published on: October 17, 2025

Area of Science:

  • Biophysics
  • Microscopy
  • Cell Biology

Background:

  • High numerical aperture immersion light microscopy is crucial for biological imaging.
  • Limitations exist in probe lifetimes and refractive index matching for cryogenic specimens.
  • Advancing imaging techniques is essential for understanding cellular dynamics.

Purpose of the Study:

  • To develop instruments and protocols for high numerical aperture immersion light microscopy on cryogenic specimens.
  • To enhance fluorescence probe lifetimes and maintain high fidelity imaging.
  • To enable correlated imaging studies with other modalities.

Main Methods:

  • Development of specialized instruments and protocols for cryogenic light microscopy.
  • Utilizing a novel cryogenic immersion fluid to minimize refractive index mismatch.
  • Integration of cryo-light imaging with soft X-ray tomography and electron microscopy.

Main Results:

  • Achieved significantly increased fluorescence probe lifetimes for protein localization studies.
  • Maintained high fidelity and spatial resolution in imaging cryogenic specimens.
  • Demonstrated applicability to both fluorescence and transmitted light microscopy, including super-resolution techniques.
  • Enabled correlated imaging of the same cryo-fixed specimen using multiple modalities.

Conclusions:

  • The developed cryogenic microscopy techniques enhance imaging capabilities for biological specimens.
  • This modality allows for the observation of dynamic cellular events followed by high-resolution imaging.
  • The methods are versatile and can be integrated with various microscopy techniques for comprehensive studies.