Confocal laser scanning fluorescence microscopy of brain microvessels: Method and preliminary results

Abstract

Chronic Hypertension alters both mechanical as well as compositional characteristics of cerebral blood vessels. We have previously determined that pial arterioles consist of two major cellular components : endothelium and smooth muscle. Smooth muscle cells are the predominant component in the arteriolar wall. The consensus of data published on the shape and dimension of vascular smooth muscle cells (VSM) is that the majority of cells are spindle shaped and of similar length and width. Changes in the morphological shape and dimension of VSM cells may occur during growth, aging or pathological conditions. In a previous study we demonstrated a digestion technique for SEM to view smooth muscle cells in arteriolar walls by removing basement membrane and arachnoid tissue covering and exposing the underlying cells. The technique proved technically unreliable, difficult to reproduce and resulted in damage to smooth muscle cells. The goal of this study was to determine if confocal microscopy techniques could offer a reliable, reproducible and safe substitute to digestion techniques in the study of cerebral smooth muscle cells.