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Published on: September 7, 2018
Identifying Active Progeny Virus Particles in Formalin-Fixed, Paraffin-Embedded Sections Using Correlative Light and
Toshiya Itoh1, Satoshi Yamada2, Isao Ohta3
1Department of Obstetrics and Gynecology, Hamamatsu University School of Medicine, Hamamatsu, Japan; Institute for NanoSuit Research, Preeminent Medical Photonics Education & Research Center, Hamamatsu University School of Medicine, Hamamatsu, Japan.
This study introduces a new method to visualize virus particles in FFPE tissue using field emission scanning electron microscopy. This technique enhances 3D imaging of viral particles, aiding in understanding virus-related diseases.
Area of Science:
- Pathology
- Microscopy
- Virology
Background:
- Immunohistochemistry on FFPE tissues is standard for identifying virus-infected cells.
- Light microscopy has limitations in resolving virus particles due to diffraction limits.
- Detecting virus particles in FFPE sections requires advanced imaging techniques.
Purpose of the Study:
- To develop a nondestructive method for observing 3D virus particles in FFPE sections.
- To overcome the resolution limitations of light microscopy for viral particle detection.
- To enable correlative light and electron microscopy for studying virus-related diseases.
Main Methods:
- Field emission scanning electron microscopy (FESEM) combined with light microscopy.
- Nondestructive 3D imaging of virus particles in FFPE sections.
- Immunostaining with specific antibodies and 3,3'-diaminobenzidine (DAB) staining, enhanced with metal solutions (gold chloride or osmium tetroxide).
Main Results:
- Successful 3D visualization of various virus particles (HPV, CMV, VZV) in FFPE sections.
- Demonstrated nondestructive imaging of FFPE sections using FESEM.
- Correlative transmission electron microscopy confirmed the presence of virus particles in FESEM-identified areas.
Conclusions:
- The combined DAB/metal staining and conductive treatment allows FESEM identification of active virus particles in FFPE sections.
- This method offers a simpler alternative to transmission electron microscopy for FFPE samples.
- Easy correlative imaging can advance the understanding of virus-related pathological mechanisms.
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