Two-photon lifetime imaging of blood and blood vessels

Abstract
We investigated the potential of two-photon excitation microscopy for the imaging in large blood vessels. Experiments were carried out on isolated rat aorta, labeled with a DNA/RNA dye. Images of the vessel wall indicated that a penetration depth of more than 200 micrometers could be reached. Moreover, blood cells and platelets inside blood vessels could be imaged through the vessel wall. Fluorescence Lifetime Imaging (FLIM) was used as a contrast mechanism for discrimination of autofluorescence from fluorescence of labeled blood cells. We were able to observe labeled blood cells through the vessel wall and identify them by their morphology and characteristic fluorescent lifetimes.

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