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Updated: Dec 29, 2025
Imaging Cell Interaction in Tracheal Mucosa During Influenza Virus Infection Using Two-photon Intravital Microscopy
Published on: August 17, 2018
Multicolor two-photon imaging of in vivo cellular pathophysiology upon influenza virus infection using the two-photon
Hiroshi Ueki1, I-Hsuan Wang1, Dongming Zhao1,2
1Division of Virology, Department of Microbiology and Immunology, Institute of Medical Science, University of Tokyo, Tokyo, Japan.
Insights
This study introduces two-photon IMPRESS, a novel imaging system for observing influenza virus infection in lungs. It allows detailed cellular-level analysis of viral pathogenesis and host responses in vivo.
Area of Science:
- Infectious Diseases
- Immunology
- Microscopy
Background:
- In vivo two-photon imaging is crucial for understanding viral pathogenesis and host immune responses.
- Analyzing influenza virus infection in the lung requires advanced imaging techniques for cellular-level insights.
Purpose of the Study:
- To describe a protocol for analyzing influenza virus-infected lungs using two-photon imaging microscopy in vivo.
- To introduce the two-photon IMPRESS (imaging pathophysiology research system) for studying respiratory pathogens.
- To provide a resource for multicolor analysis using reporter viruses, dyes, and mouse lines.
Main Methods:
- Surgical procedures for lung stabilization and two-photon imaging in vivo.
- Data analysis approaches for imaging influenza virus-infected lungs.
- Development of a database for fluorescent dyes, antibodies, and reporter mouse lines for multicolor analysis with reporter influenza virus (Color-flu).
Main Results:
- The two-photon IMPRESS system enables observation of influenza virus-infected lungs for over 4 hours in the acute phase and at least 1 hour in the lethal phase.
- The setup time for the model is approximately 30 minutes.
- The system allows for detailed, cellular-level visualization of disease progression in vivo.
Conclusions:
- Two-photon IMPRESS is a broadly applicable imaging system for analyzing respiratory pathogens in vivo.
- This methodology enhances the study of viral pathogenesis and host responses at the cellular level.
- The system provides valuable insights into disease progression during acute and lethal phases of infection.
Abstract:
In vivo two-photon imaging is a valuable technique for studies of viral pathogenesis and host responses to infection in vivo. In this protocol, we describe a methodology for analyzing influenza virus-infected lung in vivo by two-photon imaging microscopy. We describe the surgical procedure, how to stabilize the lung, and an approach to analyzing the data. Further, we provide a database of fluorescent dyes, antibodies, and reporter mouse lines that can be used in combination with a reporter influenza virus (Color-flu) for multicolor analysis. Setup of this model typically takes ~30 min and enables the observation of influenza virus-infected lungs for >4 h during the acute phase of the inflammation and at least 1 h in the lethal phase. This imaging system, which we termed two-photon IMPRESS (imaging pathophysiology research system), is broadly applicable to analyses of other respiratory pathogens and reveals disease progression at the cellular level in vivo.
