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Using Multiphoton Intravital Microscopy to Study Neutrophil Transmigration and Blood-Brain Barrier Permeability in a
Laura Bricio-Moreno1, Evelyn A Kurt-Jones2, Elizabeth W Sorensen1
1Center for Immunology and Inflammatory Diseases, Division of Rheumatology, Allergy, and Immunology, Massachusetts General Hospital and Harvard Medical School, Boston, MA, USA.
Abstract:
Multiphoton intravital microscopy (MP-IVM) is an imaging technique used for the observation of living organisms at a microscopic resolution. The tissue of interest is exposed through a window allowing imaging of cells in real time. Using MP-IVM, the temporospatial kinetics of leukocyte transendothelial migration can be visualized and quantitated using reporter mice and cell-specific fluorophore-conjugated monoclonal antibodies to track the leukocytes within and outside of vascular beds. Here we describe a method used to study neutrophil transendothelial migration and blood-brain barrier permeability in a mouse model of herpes simplex virus I (HSV) encephalitis.
Insights
Multiphoton intravital microscopy visualizes leukocyte migration in real-time. This method allows studying neutrophil behavior and blood-brain barrier changes during herpes simplex virus encephalitis.
Area of Science:
- Immunology
- Neuroscience
- Microscopy
Background:
- Multiphoton intravital microscopy (MP-IVM) enables real-time observation of biological processes in living organisms.
- Leukocyte transendothelial migration is crucial in immune responses and disease pathogenesis.
- Understanding leukocyte dynamics within the central nervous system is vital for neuroinflammatory diseases.
Purpose of the Study:
- To describe a method for studying neutrophil transendothelial migration using MP-IVM.
- To investigate blood-brain barrier permeability in a mouse model of herpes simplex virus (HSV) encephalitis.
Main Methods:
- Utilizing MP-IVM to visualize and quantify leukocyte migration in vivo.
- Employing reporter mice and fluorescently labeled antibodies for cell tracking.
- Establishing a mouse model of HSV encephalitis to study neuroinflammation.
Main Results:
- MP-IVM successfully visualized temporospatial kinetics of leukocyte migration.
- The method allowed for real-time assessment of neutrophil extravasation.
- Changes in blood-brain barrier permeability were observed in the HSV encephalitis model.
Conclusions:
- MP-IVM is a powerful tool for studying leukocyte migration in neurological infections.
- This technique provides valuable insights into neuroinflammatory processes and blood-brain barrier integrity.
- The described method facilitates research into HSV encephalitis pathogenesis.
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