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Comparison of protein A-gold and ferritin immunoelectron microscopy of Semliki Forest virus in mouse brain using a
Abstract:
Processing tissue for transmission electron microscopy by standard laboratory methods can take two to three days. This makes the development of new techniques time consuming and generally restricts the use of the electron microscope in routine diagnostic work. The possibility of viewing tissue with the electron microscope five hours after sampling using rapid processing techniques is presented. The morphology of the tissue appears undamaged with cell and organelle ultrastructures being readily recognized, as is the presence of virus and its replicating stages. When combined with immunoelectron microscopy a rapid labeling protocol is possible. We have used the technique to develop protein A-gold (6 and 16 nm particles) and ferritin immunoelectron microscopic techniques to demonstrate viral antigens in brain cell cultures and brain tissue from mice infected with Semliki Forest virus.
Insights
This study presents a rapid tissue processing technique for transmission electron microscopy, reducing sample viewing time from days to hours. This method preserves tissue morphology and enables quick identification of viruses and antigens, aiding diagnostic work.
Area of Science:
- Electron Microscopy
- Virology
- Cell Biology
Background:
- Standard transmission electron microscopy (TEM) tissue processing is time-consuming (2-3 days).
- This delay hinders new technique development and routine diagnostic applications of TEM.
- Rapid processing is needed to expedite electron microscopy analysis.
Purpose of the Study:
- To present a rapid tissue processing technique for TEM.
- To reduce sample analysis time to five hours post-sampling.
- To enable rapid immunoelectron microscopy protocols.
Main Methods:
- Developed a rapid processing technique for transmission electron microscopy.
- Utilized protein A-gold and ferritin for immunoelectron microscopy labeling.
- Applied the technique to Semliki Forest virus-infected mouse brain cell cultures and tissue.
Main Results:
- Tissue morphology and ultrastructure were preserved.
- Cell and organelle structures were readily recognized.
- Viral presence and replication stages were identifiable.
- Rapid immunoelectron microscopy successfully demonstrated viral antigens.
Conclusions:
- The rapid processing technique significantly reduces TEM analysis time.
- This method maintains tissue integrity for ultrastructural analysis.
- It facilitates rapid identification of viral components and antigens, enhancing diagnostic capabilities.