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Polyester film as an advantageous tool for light and ultrastructural examination of single cells infected with
Abstract:
The use of marked transparent polyester film as a support of cell monolayer, smears or sections (cryostat or paraffin), makes possible to examine the same cell preselected in the light microscope by electron microscope (EM). Consequently, this technique enables to use histological or immunohistological investigations (using immunofluorescence or peroxidase (Px) labelling) by light microscopy before examination in EM. As example we used herpes simplex virus (HSV) type 1-infected cells grown on the above mentioned polyester film. Visualization of the antigen with hyperimmune serum and Px-labelled antiserum was followed by examination of selected cells in EM, where viral particles were clearly visible.
Insights
Transparent polyester film allows researchers to examine the same cells using both light and electron microscopy. This technique facilitates correlative light and electron microscopy for detailed cellular and viral particle visualization.
Area of Science:
- Cell Biology
- Microscopy Techniques
- Virology
Background:
- Correlative microscopy requires techniques that allow sequential examination of the same biological sample.
- Traditional methods may not efficiently bridge light and electron microscopy for cellular analysis.
Purpose of the Study:
- To introduce a novel method using marked transparent polyester film for correlative light and electron microscopy.
- To demonstrate the utility of this technique for visualizing viral particles in infected cells.
Main Methods:
- Cells were cultured on marked transparent polyester film.
- Immunohistochemical staining (peroxidase labeling) was performed for light microscopy.
- The same cells were subsequently examined using electron microscopy.
Main Results:
- The polyester film supported cell monolayers, smears, and sections for both microscopy types.
- Immunohistological investigations identified specific antigens.
- Electron microscopy clearly visualized viral particles in herpes simplex virus type 1-infected cells.
Conclusions:
- Marked transparent polyester film is an effective support for correlative light and electron microscopy.
- This method enables precise localization and detailed ultrastructural analysis of cellular components and pathogens.