Abstract: Disclosed herein are methods of vascular and structural imaging using imaging systems and methods described, the methods comprising producing an image of the vasculature or structure by imaging fluorescence using an imaging system, the system comprising: i) one or more detectors configured to form a fluorescence image of the sample and form a visible image of the sample; ii) a light source configured to emit an excitation light to induce fluorescence from the sample; and iii) a plurality of optics arranged to: direct the excitation light toward the sample; and direct a fluorescent light and a visible light from the sample to the detector; wherein the excitation light and the fluorescence light are directed substantially coaxially.