Photothermal imaging of moving cells in lymph and blood flow in vivo

Abstract

The in vivo capabilities of a new, integrated optical system for studying lymph and blood flow were explored, including imaging of moving red and white blood cells. This system combined transmission microscopy with different dual-beam photothermal (PT) techniques, such as PT imaging, PT thermolens method, and PT deflection velocimetry. All of these PT techniques are based on irradiation of rat mesenteric microvessels with a short laser pulse and on detection of temperature-dependent variations of the refractive index with a second, probe laser beam. In general, the concept of in vivo PT flow cytometry was developed, with a focus on real-time monitoring of moving blood cells in their natural states without labeling (e.g., fluorescent), including obtaining PT images of the cells and determining their flow velocity and response to different interventions. Preliminary experiments revealed many potential applications of this integrated system: (1) quantitation of lymph and blood flow without probes; (2) imaging of moving red and white blood cells; (3) visualization and tracking of PT nanoprobes and sensitizers; (4) comparison of laser-tissue interactions in vivo and in vitro, especially optimization of laser treatment of vascular lesions (port-wine stains, lymphatic malformations, etc.); and (5) determination of the link between in vitro and in vivo cytotoxicity studies.