Craniula: A cranial window technique for prolonged imaging of brain surface vasculature with simultaneous adjacent intracerebral injectionReport as inadecuate




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Fluids and Barriers of the CNS

, 12:24

First Online: 27 October 2015Received: 16 April 2015Accepted: 19 October 2015

Abstract

BackgroundImaging of the brain surface vasculature following inflammatory insults is critical to study structural and functional changes in the living brain under normal and pathological conditions. Although there have been published reports relating to the changes that occur in the blood brain barrier BBB during the inflammatory process, the ability to visualize and track such changes in vivo and over time has proven to be problematic. Different techniques have been used to achieve visualization of pial vessels, but the approach has limits, which can jeopardize the well-being of the animals. Development of the cranial window technique provided a major advance in the acquisition of live images of the brain vasculature and its response to different insults and treatments.

MethodsWe describe in detail a protocol for delivery of a localized inflammatory insult to the mouse brain via a craniula cranial window and adjacent cannula and subsequent imaging of the mouse brain vasculature by intravital microscopy and two-photon laser scanning microscopy. The surgical implantation of the craniula can be completed in 30-45 min and images can be acquired immediately and for several months thereafter. The technique is minimally invasive and permits serial injections directly to the brain, thereby allowing longitudinal imaging studies. The craniula technique permits the study of structural and functional changes of the BBB following inflammatory insult and as such has wide application to neuroscience research.

KeywordsBlood Brain Barrier Leukocyte–endothelial interaction Intravital microscopy 2-photon microscopy Viviana Zuluaga-Ramirez and Slava Rom contributed equally to this work

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Author: Viviana Zuluaga-Ramirez - Slava Rom - Yuri Persidsky

Source: https://link.springer.com/



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