Dynamic in vivo imaging of microvasculature and perfusion by miniaturized confocal laser microscopy.

Abstract

INTRODUCTION: Microvasculature and associated pathologies mandate dynamic imaging. We evaluated a novel miniaturized confocal laser scanning probe for in vivo visualization of blood vessels, blood flow, cell tracking and perfusion in both healthy rodents and disease models. METHODS: The hand-held confocal microscopy system allowed a 500- to 2,400-fold magnification at a dynamically variable imaging depth. Different intravital stains were used alone or in combination for tissue, nuclear, plasma and vascular endothelial cell staining and for blood flow visualization, and targeted staining for individual cell populations. RESULTS: Precision optical sectioning yielded high-resolution images in vivo. Leucocyte-endothelium interactions in brain microvasculature were followed in serial sections. A microthrombosis was identified after sequential injection of FITC-labelled erythrocytes, FITC-dextran and acriflavine. Glomerular alterations were visualized in the MRL/lprmouse model of lupus nephritis. DISCUSSION: Intravital confocal microscopy with a miniaturized hand-held probe combines high-resolution subsurface imaging in real time for dynamic visualization of vessels, cells, blood flow and associated pathologies, permitting a truly comprehensive vascular imaging in vivo at the cellular level.