Optimization of Multiphoton Imaging for Quantification of Cerebral Microvessel Permeability in Murine Ischemic Stroke

Chloe G. Hernandez* ¹, Briana L. Clary ^2,3, and Candice M. Brown ^2,3
Departments of ¹Biology and ²Neuroscience, ³Center for Basic and Translational Stroke Research and Rockefeller Neuroscience Institute, West Virginia University School of Medicine, Morgantown, WV

Presentation No.: 93

Assigned Category (Presentation Format): Neuroscience (Poster Presentations)

Student’s Major: Neuroscience and Psychology

Ischemic stroke encompasses ~85% of all strokes. Multiphoton (MP) microscopy is commonly used for acute and chronic imaging of brain tissue through a cranial window preparation. Fluorescently labeled cerebral microvessels are employed in MP imaging to assess real-time changes in blood-brain permeability, vascular remodeling, and inflammation. Since MP imaging is a new technique that will be implemented in our laboratory, the goal of this study was to optimize imaging parameters for MP imaging to study longitudinal changes in ischemic stroke. First, we optimized cranial window placement surgery to facilitate chronic MP imaging. We will then use the photothrombotic stroke (PTS) model, which mimics a permanent ischemic stroke, and fluorescent molecular dyes to elucidate the cerebral microvascular changes that occur in ischemic stroke. Another tool we have developed facilitate imaging of fluorescent cerebral microvessels is a mouse that expresses the tdTomato fluorescent marker in endothelial cells. Taken together, these tools will allow us to study the role of tissue-nonspecific alkaline phosphatase as a regulator of cerebral microvessel permeability in ischemic stroke.

Funding:

Program/mechanism supporting research/creative efforts: Other