Related Experiment Video
Updated: May 24, 2025
08:37
Visualization of Organelles In Situ by Cryo-STEM Tomography
Published on: June 23, 2023
2.0K
Cryo-STEM tomography for cell biology using thick lamella
Kazuhiro Aoyama1,2, Hiroko Takazaki3, Misaki Arie3
1NanoPort Japan Application Laboratory, Thermo Fisher Scientific, Shinagawa Seaside West Tower 1F, 4-12-2 Higashi-Sinagawa, Shinagawa-ku, Tokyo 140-0002, Japan.
Microscopy (Oxford, England)
|March 4, 2025
Summary
Scanning transmission electron microscopy (STEM) tomography overcomes thickness limitations in electron tomography (ET) for cell biology. This technique enables detailed 3D reconstruction of thicker biological specimens, advancing structural studies.
Area of Science:
- Cell Biology
- Microscopy
- Structural Biology
Background:
- Electron tomography (ET) is crucial for cell biology structural studies.
- Specimen thickness is a major limitation for traditional ET.
- Scanning transmission electron microscopy (STEM) offers potential advantages.
Purpose of the Study:
- To evaluate the utility of cryo-STEM tomography for imaging thicker biological specimens.
- To demonstrate the capability of cryo-STEM tomography for 3D reconstruction of cellular structures.
Main Methods:
- Utilized focused ion beam (FIB) slicing for cryo-specimen preparation.
- Employed cryo-STEM tomography for high-resolution imaging.
- Performed 3D reconstruction of cellular organelles.
Main Results:
- Successfully reconstructed relatively thick specimens (300-500 nm).
- Clearly visualized organelles, including mitochondria and the nuclear membrane.
- Demonstrated the feasibility of cryo-STEM tomography for thick samples.
Conclusions:
- Cryo-STEM tomography is a promising technique for overcoming specimen thickness limitations in cell biology.
- Enables detailed structural analysis of organelles in thicker biological samples.
- Advances the field of structural cell biology through improved imaging capabilities.
Related Concept Videos
01:07Electron Microscope Tomography and Single-particle Reconstruction
2.3K
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...
Electron Tomography
Electron tomography can be performed either in TEM or STEM (scanning transmission...
2.3K
01:28Cryo-electron Microscopy
3.2K
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...
3.2K