Intravital Widefield Fluorescence Microscopy of Pulmonary Microcirculation in Experimental Acute Lung Injury Using a
Stefan Hall1, Sufyan Faridi2, Irene Euodia3
1Department of Physiology and Biophysics, Dalhousie University.
Journal of Visualized Experiments : Jove
|April 25, 2022
Summary
This study presents a novel intravital microscopy method for visualizing leukocyte behavior in the lungs during acute lung injury (ALI). The technique overcomes imaging challenges to reveal real-time immune cell interactions in pulmonary microcirculation.
Area of Science:
- Immunology
- Physiology
- Microscopy
Background:
- Intravital imaging of leukocyte-endothelial interactions is crucial for understanding immune-mediated diseases.
- Studying respiratory pathologies like acute lung injury (ALI) and acute respiratory distress syndrome (ARDS) in vivo is challenging due to lung accessibility and motion artifacts.
Purpose of the Study:
- To describe a protocol for intravital fluorescence microscopy to visualize leukocyte-endothelial interactions in the pulmonary microcirculation.
- To adapt existing intravital microscopy techniques for studying ALI in a live animal model.
Main Methods:
- Utilized an in vivo lung imaging system and a 3-D printed intravital microscopy platform for anesthetized mice.
- Employed widefield fluorescence microscopy to observe leukocyte adhesion, rolling, and capillary function in real-time.
- Minimized confounding lung injury during the procedure.
Main Results:
- Successfully visualized leukocyte-endothelial interactions in the pulmonary microcirculation during an experimental ALI model.
- Demonstrated the ability to study dynamic cellular processes in the lung with high resolution.
- Validated the use of the specialized imaging platform for stabilizing lung tissue.
Conclusions:
- The developed intravital microscopy protocol enables real-time study of leukocyte behavior in the pulmonary microcirculation during ALI.
- This method overcomes significant technical hurdles in lung intravital imaging.
- The protocol can be adapted for investigating various other lung pathologies and physiological processes.
