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Related Concept Videos

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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A Microfluidic Approach for the Study of Ice and Clathrate Hydrate Crystallization
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High-Speed Video Cryomicroscopy for Measurement of Intracellular Ice Formation Kinetics.

Jens O M Karlsson1

  • 1Department of Mechanical Engineering, Villanova University, Villanova, PA, USA. jens.karlsson@villanova.edu.

Methods in Molecular Biology (Clifton, N.J.)
|August 16, 2020
PubMed
Summary

High-speed video cryomicroscopy accurately detects intracellular ice formation kinetics. This method improves cryopreservation by providing precise data on freezing events in cells and tissues.

Keywords:
CryomicroscopeCumulative hazardCumulative probabilityFlashingHigh-speed imagingIntracellular ice formationKineticsNelson-Aalen estimatorNucleationUltra-slow motion

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Area of Science:

  • Cryobiology
  • Biophysics
  • Cell Biology

Background:

  • Accurate data on intracellular ice formation kinetics is crucial for developing effective cryopreservation techniques.
  • Conventional methods for detecting intracellular freezing can lead to significant errors in kinetic estimations.

Purpose of the Study:

  • To describe the setup and operation of a high-speed video cryomicroscope system.
  • To provide protocols for imaging and analyzing intracellular ice crystallization events and kinetics in cell populations.

Main Methods:

  • Utilizing temporally resolved imaging (sub-millisecond video recording) for accurate detection of intracellular ice formation and growth.
  • Implementing stochastic analysis for quantifying ice formation kinetics within a cell population.
  • Detailing best practices for temperature profile design, sample preparation, and video acquisition parameters.

Main Results:

  • High-speed video cryomicroscopy offers a more accurate method for assessing intracellular ice formation compared to conventional assays.
  • The described protocols enable detailed quantitative analysis of ice crystallization events.
  • The study provides a robust framework for reliable cryopreservation research.

Conclusions:

  • High-speed video cryomicroscopy is a superior technique for studying intracellular ice formation kinetics.
  • Accurate kinetic data is essential for optimizing cryopreservation protocols to minimize cell damage.
  • The presented methods and best practices enhance the reliability and precision of cryobiology research.