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A differential in vivo two photon microscopy method for quantitative imaging of dye extravasation in mouse cortex

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Abstract

An improved two-photon microscopy technique for imaging and quantifying blood brain barrier (BBB) leakage in the cortex of mice is presented. Local BBB leakage was induced using synchrotron X-ray micro-beams. Two fluorescent dyes were simultaneously infused intravenously, one of high molecular weight (FITC-dextran) and one of low molecular weight (sulforhodamine B, SRB). By separation of the two fluorescence spectra, and matched subtraction, images were obtained in which the vasculature was invisible and the low molecular weight dye in the parenchyma could be imaged with high resolution. An algorithm is presented for quantifying the extravasation without the need of vessel recognition.

© 2006 Optical Society of America

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