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Related Concept Videos

Immunogold Electron Microscopy01:20

Immunogold Electron Microscopy

Immunoelectron microscopy utilizes immunogold labeling of endogenous proteins with specific antibodies to detect and localize these proteins in cells and tissues. The procedure provides insights into the distribution and quantification of protein under different stimulation conditions offering clues about their functions. Conjugating highly electron-dense gold particles with primary or secondary antibodies allow antigen detection on and within cells, with high resolution and specificity.
Preparation of Samples for Electron Microscopy01:20

Preparation of Samples for Electron Microscopy

To be visualized by an electron microscope, either transmission or scanning, biological samples need to be fixed (stabilized) so the electron beam does not destroy them and dried thoroughly (desiccated/dehydrated) so the vacuum does not affect them. Fixation needs to be done as quickly as possible because the sample properties will start changing as soon as it is removed from its natural environment. For example, in a tissue sample, the oxygen levels begin decreasing, causing an altered...
Transmission Electron Microscopy01:15

Transmission Electron Microscopy

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Electron Microscope Tomography and Single-particle Reconstruction01:07

Electron Microscope Tomography and Single-particle Reconstruction

Transmission electron microscopy (TEM) can be used to determine the 3D structure of biological samples with the help of techniques such as electron microscope tomography and single-particle reconstruction. While single-particle reconstruction can examine macromolecules and macromolecular complexes in vitro conditions only, tomography permits the study of cell components or small cells in vivo.
Electron Tomography
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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...

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Updated: May 21, 2026

Optimized Negative Staining: a High-throughput Protocol for Examining Small and Asymmetric Protein Structure by Electron Microscopy
09:37

Optimized Negative Staining: a High-throughput Protocol for Examining Small and Asymmetric Protein Structure by Electron Microscopy

Published on: August 15, 2014

Imaging Lipid-Associated Macromolecular Structures by Optimized Negative-Staining Transmission Electron Microscopy.

Jianfang Liu1, Meng Zhang1, Gang Ren2

  • 1Molecular Foundry, Lawrence Berkeley National Laboratory, Berkeley, CA, USA.

Methods in Molecular Biology (Clifton, N.J.)
|May 19, 2026
PubMed
Summary
This summary is machine-generated.

Researchers optimized negative staining (NS) for imaging macromolecule-lipid interactions. This new protocol minimizes artifacts, yielding high-quality, near-native images for accurate structural analysis.

Keywords:
IPETIndividual-particle electron tomographyLipoprotein morphologyLipoprotein structureMacromoleculer structureNegative-staining electron microscopyOpNSOptimized negative-staining protocolSingle molecule 3D structureTransmission electron microscopy

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Last Updated: May 21, 2026

Optimized Negative Staining: a High-throughput Protocol for Examining Small and Asymmetric Protein Structure by Electron Microscopy
09:37

Optimized Negative Staining: a High-throughput Protocol for Examining Small and Asymmetric Protein Structure by Electron Microscopy

Published on: August 15, 2014

Method to Visualize and Analyze Membrane Interacting Proteins by Transmission Electron Microscopy
10:49

Method to Visualize and Analyze Membrane Interacting Proteins by Transmission Electron Microscopy

Published on: March 5, 2017

Variations on Negative Stain Electron Microscopy Methods: Tools for Tackling Challenging Systems
06:06

Variations on Negative Stain Electron Microscopy Methods: Tools for Tackling Challenging Systems

Published on: February 6, 2018

Area of Science:

  • Structural biology
  • Biophysics
  • Biochemistry

Background:

  • Macromolecules like proteins interact with lipids, crucial for biological processes.
  • Structural determination of lipid-bound proteins is challenging due to sample heterogeneity.
  • Transmission electron microscopy (TEM) visualizes individual particles, but negative staining (NS) can cause artifacts.

Purpose of the Study:

  • To develop an optimized negative staining (OpNS) protocol for imaging macromolecule-lipid interactions.
  • To overcome limitations of conventional NS, such as artifact formation in lipoproteins.
  • To enable accurate structural analysis of macromolecules in their native lipid-bound states.

Main Methods:

  • Refinement of existing negative staining protocols.
  • Validation against cryo-electron microscopy (cryo-EM) of vitrified lipoproteins.
  • Application of optimized protocol for imaging macromolecule-lipid complexes.

Main Results:

  • The optimized negative staining (OpNS) protocol minimizes artifacts, producing high-quality, near-native images.
  • OpNS enables more accurate structural analysis compared to conventional NS.
  • The protocol supports advanced techniques like individual-particle electron tomography (IPET) for 3D reconstruction.

Conclusions:

  • The OpNS protocol offers a robust and efficient method for imaging macromolecules interacting with lipids.
  • This advancement facilitates a deeper understanding of macromolecule-lipid interactions at a structural level.
  • The improved imaging technique supports structural biology and biophysical studies of membrane-associated molecules.