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Coated Glass Slides TACAS Are Applicable to Heat-Assisted Immunostaining and In Situ Hybridization at the Electron
Takahiro Matsui1, Takanori Onouchi2, Kazuya Shiogama2
1Student, the Faculty of Medical Technology, Fujita Health University School of Health Sciences , Toyoake, Aichi, Japan ; Department of Pathology, Fujita Health University School of Medicine , Toyoake, Aichi, Japan ; Present address: Laboratory Medicine, Toyota Kosei Hospital , Toyota, Aichi, Japan.
Abstract:
We performed pre-embedding electron microscopic study for visualizing the antigen and genome of severe fever with thrombocytopenia syndrome (SFTS) virus in the cytoplasm of macrophages of the human splenic red pulp, both requesting preheating treatment of sections. To pursue this, coated glass slides with unique characteristics are needed. Namely, during staining they must prevent detaching off sections, but after staining the sections must be transferred to epoxy resin. Aminopropyltriexoxysilane-coated glass slides, widely used for immunostaining, were resistant to transfer to epoxy resin. In contrast, coated glass slides designated as Thinlayer Advanced Cytology Assay System (TACAS) were suitable for this purpose. The technique is also applicable to the coated glass slide-requiring cytology practice, in which immunocytochemical evaluation is needed after cell transfer to another glass slide.
Insights
This study introduces a new method for visualizing severe fever with thrombocytopenia syndrome (SFTS) virus using specialized coated glass slides. The Thinlayer Advanced Cytology Assay System (TACAS) slides enable antigen and genome visualization in macrophages.
Area of Science:
- Electron Microscopy
- Virology
- Cell Biology
Background:
- Visualizing viral components within host cells is crucial for understanding disease pathogenesis.
- Pre-embedding electron microscopy requires specialized sample preparation techniques.
- Current methods using aminopropyltriexoxysilane-coated slides are unsuitable for epoxy resin transfer.
Purpose of the Study:
- To develop and validate a novel method for visualizing severe fever with thrombocytopenia syndrome (SFTS) virus antigen and genome.
- To identify suitable coated glass slides for pre-embedding electron microscopy requiring epoxy resin transfer.
- To adapt this technique for immunocytochemical evaluation after cell transfer.
Main Methods:
- Pre-embedding electron microscopy was performed on human splenic red pulp macrophages.
- Sections underwent preheating treatment for antigen and genome visualization.
- Thinlayer Advanced Cytology Assay System (TACAS) coated glass slides were utilized and compared to aminopropyltriexoxysilane-coated slides.
Main Results:
- Aminopropyltriexoxysilane-coated slides prevented section transfer to epoxy resin.
- TACAS-coated glass slides facilitated successful transfer of stained sections to epoxy resin.
- The developed technique enabled visualization of SFTS virus in macrophage cytoplasm.
Conclusions:
- TACAS-coated glass slides are suitable for pre-embedding electron microscopy requiring epoxy resin transfer.
- This method allows for the visualization of SFTS virus components in macrophages.
- The technique is adaptable for other cytology practices needing post-transfer immunocytochemical evaluation.
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