Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Cryo-electron Microscopy01:28

Cryo-electron Microscopy

3.3K
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...
3.3K

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Using <i>spIsoNet</i> to address the preferred-orientation problem in cryoEM reconstructions.

bioRxiv : the preprint server for biology·2026
Same author

Structure and potential role of T6SS effector PdpC in Francisella tularensis intracellular lifestyle.

Communications biology·2026
Same author

Study on removal of heavy metal ions from electroplating wastewater using red mud leachate.

Journal of environmental management·2026
Same author

Transport mechanism of the SLC4 proteins-Lessons from recent structural and computational studies.

The Journal of biological chemistry·2026
Same author

Toxoplasma IMC1 is a central component of the subpellicular network and plays critical roles in parasite morphology, replication, and infectivity.

PLoS pathogens·2026
Same author

Atomic clarity: how structural biology is shaping blood-stage malaria vaccines.

Transactions of the Royal Society of Tropical Medicine and Hygiene·2026

Related Experiment Video

Updated: Jun 30, 2025

Preparation of Sample Support Films in Transmission Electron Microscopy using a Support Floatation Block
06:10

Preparation of Sample Support Films in Transmission Electron Microscopy using a Support Floatation Block

Published on: April 8, 2021

4.5K

Theoretical framework and experimental solution for the air-water interface adsorption problem in cryoEM.

Joon S Kang1,2, Xueting Zhou1, Yun-Tao Liu1,3

  • 1Department of Microbiology, Immunology & Molecular Genetics, University of California, Los Angeles (UCLA), Los Angeles, CA 90095, USA.

Biophysics Reports
|March 22, 2024
PubMed
Summary

Cryo-electron microscopy (cryoEM) struggles with particle loss at the air-water interface. Surfactants reduce surface tension, minimizing particle loss and enabling high-resolution cryoEM structure determination.

Keywords:
Air–water interface adsorptionCryoEMCryoETSample preparationSurface energySurfactant

More Related Videos

Author Spotlight: Enhancing CryoEM Resolution Using Graphene-Coated Grids
06:53

Author Spotlight: Enhancing CryoEM Resolution Using Graphene-Coated Grids

Published on: September 8, 2023

3.1K
Optimizing Sample Preparation for Cryogenic Electron Microscopy
06:32

Optimizing Sample Preparation for Cryogenic Electron Microscopy

Published on: April 11, 2025

400

Related Experiment Videos

Last Updated: Jun 30, 2025

Preparation of Sample Support Films in Transmission Electron Microscopy using a Support Floatation Block
06:10

Preparation of Sample Support Films in Transmission Electron Microscopy using a Support Floatation Block

Published on: April 8, 2021

4.5K
Author Spotlight: Enhancing CryoEM Resolution Using Graphene-Coated Grids
06:53

Author Spotlight: Enhancing CryoEM Resolution Using Graphene-Coated Grids

Published on: September 8, 2023

3.1K
Optimizing Sample Preparation for Cryogenic Electron Microscopy
06:32

Optimizing Sample Preparation for Cryogenic Electron Microscopy

Published on: April 11, 2025

400

Area of Science:

  • Structural Biology
  • Biophysics
  • Biochemistry

Background:

  • Cryo-electron microscopy (cryoEM) is a powerful technique for determining atomic structures of biological complexes.
  • Many complexes exhibit problematic behavior on cryoEM grids, such as preferential orientation or aggregation, hindering structure determination.
  • The underlying reasons for these issues, particularly particle loss, are not well understood.

Purpose of the Study:

  • To develop a theoretical explanation for particle misbehavior in cryoEM.
  • To propose and validate the use of surfactants as a solution to improve cryoEM grid preparation.
  • To demonstrate the utility of this approach for determining high-resolution structures of challenging biological complexes.

Main Methods:

  • Theoretical formulation to explain particle behavior at the air-water interface (AWI).
  • Cryogenic electron tomography (cryoET) to visualize particle distribution on cryoEM grids.
  • Single-particle cryoEM to assess structural integrity after surfactant treatment.
  • Application to GroEL and the ClC-1 channel membrane protein.

Main Results:

  • Particles preferentially migrate to the AWI to minimize surface energy.
  • Reducing surface tension with surfactants significantly decreased particle accumulation at the AWI.
  • Suitable surfactants did not compromise the structural integrity of biological complexes.
  • Near-atomic structure of the ClC-1 channel was determined using this method.

Conclusions:

  • Particle migration to the AWI is a key factor in cryoEM grid preparation challenges.
  • Surfactants offer a practical, generalizable solution to mitigate AWI adsorption issues in cryoEM.
  • This approach facilitates high-resolution structure determination of difficult biological samples, including membrane proteins.