2015 Volume 26 Issue 2 Pages 99-105
A precise understanding of the cellular interplay in the neurovascular unit is fundamental to develop a way of preventing neurodegenerative disorders resulting from cerebrovascular disturbance. A new imaging technique using in vivo two-photon microscopy with a variety of animal models that genetically express fluorescence proteins in the cells enables us to explore the structural and functional plastic changes in the neurovascular unit. Here we report the optical imaging and analytical techniques to visualize and quantify the neurovascular structures and functions. For imaging of brain microvasculature, a capillary length and diameter was automatically measured and their three-dimensional images were reconstructed by using custom-written matlab software. For brain cell imaging, either neurons or glias were fluorescently labeled, and their morphological parameters were determined along with a location of the measured cells. In addition to the static imaging, a dynamic scanning method allows for acquiring a functional property of the cerebral blood flow and/or intracellular calcium concentrations. Finally, a future work should focus on a development of the imaging and quantification methods for cellular environment and genetic and/or protein expressions. Further technical improvements with in vivo two-photon microscopy with a variety of fluorescent proteins will provide a powerful tool for understanding the long-term neurovascular plasticity in health and disease.
