Related Experiment Videos
Three-dimensional high voltage electron microscopy of thick biological specimens
1Department of Anatomy and Cell Biology, Shinshu University School of Medicine, Matsumoto, Japan. nagatas@po.cnet.ne.jp
Micron (Oxford, England : 1993)
|November 9, 2000
Summary
High voltage electron microscopy enables three-dimensional visualization of cell organelles in thick biological specimens. This technique allows detailed stereological analysis of cellular structures and their relationships.
Area of Science:
- Cell Biology
- Microscopy
- Biophysics
Background:
- High voltage electron microscopy (HVEM) offers unique capabilities for imaging thick biological specimens.
- Traditional electron microscopy methods are limited in visualizing the three-dimensional (3D) ultrastructure of intact cells and tissues.
Purpose of the Study:
- To review and present procedures for observing 3D structures of cell organelles in thick biological specimens using HVEM.
- To demonstrate the utility of HVEM for stereological analysis of cellular ultrastructure.
Main Methods:
- Utilized HVEM (400-1000kV) to examine whole mount cultured cells and thick tissue sections (1-2 µm).
- Employed histochemical staining for specific marker enzymes (e.g., G-6-Pase, TPPase, cytochrome oxidase) and radioautography.
- Recorded stereo-pair micrographs and analyzed them using image analysis software for 3D reconstruction.
Main Results:
- Successfully visualized 3D structures of organelles like endoplasmic reticulum, Golgi apparatus, mitochondria, lysosomes, and peroxisomes.
- Demonstrated the localization of marker enzymes and radiolabeled compounds within specific organelles.
- Revealed the structural relationships between various cell organelles in 3D.
Conclusions:
- Developed and validated procedures for 3D stereological analysis of cell organelles using HVEM.
- HVEM provides a powerful tool for understanding cellular ultrastructure and organelle dynamics in complex biological samples.