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Related Concept Videos

The Structure of Intermediate Filaments01:19

The Structure of Intermediate Filaments

The intermediate filaments are one of three widely studied cytoskeletal filaments. They are so named as their diameter (10 nm) is in between that of microfilaments (7 nm) and the microtubules (25 nm).  These filaments are highly stable and can remain intact when exposed to high salt concentrations and detergents. These filaments are responsible for providing stability and mechanical support to the cells. They also help in cell adhesion and maintaining tissue integrity.
Intermediate filaments...
Studying the Cytoskeleton01:17

Studying the Cytoskeleton

The cytoskeletal architecture can be studied using different microscopic and biochemical techniques. Electron microscopy was instrumental in discovering the cytoskeletal architecture around the 1960s, which allowed obtaining structural information at a high-resolution level. However, the sample preparation procedure often limits this ability in biological samples. Several protocols have been developed over the years to optimize sample preparation. In one of the protocols known as rotary...
Formation of Intermediate Filaments00:57

Formation of Intermediate Filaments

Intermediate filaments are cytoskeletal proteins with higher tensile strength and flexibility than microfilaments and microtubules. Unlike the other two cytoskeletal proteins, intermediate filament formation lacks the enzymatic activity to hydrolyze nucleotides like ATP and GTP to generate energy for polymerization. Therefore, the formation of intermediate filaments is multistep self-assembly. The involvement of any accessory proteins in intermediate filament formation has not yet been reported.
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...
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
Electron tomography can be performed either in TEM or STEM (scanning transmission...
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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Imaging Intermediate Filaments and Microtubules with 2-dimensional Direct Stochastic Optical Reconstruction Microscopy
14:23

Imaging Intermediate Filaments and Microtubules with 2-dimensional Direct Stochastic Optical Reconstruction Microscopy

Published on: March 6, 2018

Three-dimensional cryo-electron microscopy on intermediate filaments.

Robert Kirmse1, Cédric Bouchet-Marquis, Cynthia Page

  • 1The Boulder Laboratory for 3-D Microscopy of Cells, University of Colorado at Boulder, Boulder, Colorado 80309-0347, USA.

Methods in Cell Biology
|September 28, 2010
PubMed
Summary
This summary is machine-generated.

Intermediate filaments (IFs) are diverse cytoskeletal components. Cryo-electron microscopy (cryo-EM) and cryo-electron tomography (cryo-ET) offer new insights into their structure and assembly.

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Last Updated: Jun 8, 2026

Imaging Intermediate Filaments and Microtubules with 2-dimensional Direct Stochastic Optical Reconstruction Microscopy
14:23

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Published on: March 6, 2018

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Area of Science:

  • Cell Biology
  • Structural Biology

Background:

  • Intermediate filaments (IFs) are a key component of the metazoan cytoskeleton, alongside microtubules and actin filaments.
  • Unlike microtubules and actin, IFs exhibit significant diversity, lack polarity, and are less structurally resolved due to functional variability and intrinsic properties.
  • The antiparallel α-helical coiled-coil structure of IF protofibrils hinders high-resolution structural determination by traditional methods like crystallography.

Purpose of the Study:

  • To evaluate the applicability of cryo-electron microscopy (cryo-EM) and cryo-electron tomography (cryo-ET) for studying intermediate filament (IF) structure.
  • To explore novel structural features and assembly properties of IFs using advanced cryo-EM techniques.

Main Methods:

  • Application of cryo-electron microscopy (cryo-EM) for high-resolution imaging of intermediate filaments (IFs).
  • Utilization of cryo-electron tomography (cryo-ET) to obtain three-dimensional reconstructions of IFs.
  • Analysis of negatively stained electron microscopy and X-ray diffraction data for fragmented IF components.

Main Results:

  • Cryo-EM has revealed previously unobserved features within intermediate filaments (IFs).
  • Cryo-ET provides a 3-D perspective, detailing protofilament arrangement and pathways within IF assemblies.
  • These advanced techniques overcome limitations of traditional methods in resolving IF macromolecular structures.

Conclusions:

  • Cryo-electron microscopy and cryo-electron tomography are powerful tools for advancing the structural understanding of intermediate filaments (IFs).
  • These methods offer unprecedented views into the complex organization and assembly of IFs, complementing existing structural data.