Measurement of cerebral microvessel diameters after embolic stroke in rat using quantitative laser scanning confocal microscopy

Abstract

Perfusion of the microcirculation after stroke is central to preserving neuronal function and improving functional outcome. Using laser scanning confocal microscopy (LSCM) and a novel computer software program, MIRAGE, we measured a reduction in cerebral microvessel diameter after 1 and 4 h of embolic middle cerebral artery (MCA) occlusion. The methodology used by MIRAGE for measurement of blood vessel diameter involves a linearly applied spherical inflation technique. Three-dimensional quantitative analysis revealed a significant ( P<0.05) mean 10–12% reduction in vessel diameter in the ipsilateral cortex when compared to the homologous region in the contralateral hemisphere. This reduction was seen only in the cortex and not the striatum. A larger reduction in mean vessel diameter in the ipsilateral cortex, 16–30% ( P<0.05) was observed when compared to sham control rats. These results are in strong agreement with other studies using different stroke models and imaging techniques. Our work represents a novel application of LSCM technology to the three dimensional investigation of microvessel diameter changes in acute stroke and identifies its potential as an important tool for investigation of cerebral pathology.