Identification of rolling circulating tumor cells using photoacoustic time-of-flight method

Abstract

Existing optical techniques for in vivo measurement of blood flow velocity are not quite applicable for determination of velocity of individual cells or nanoparticles. A time-of-flight photoacoustic (PA) technique can solve this problem by measuring the transient PA signal width, which is related to the cell velocity passing the laser beam. This technique was demonstrated in vivo using an animal (mouse) model by estimating the velocity of nanoparticles, and red and white blood cells labeled with conjugated gold nanorods (GNRs) in the bloodstream. Here we describe the features and the parameters of novel modifications to the PA time-of-flight method and its new application for real-time monitoring of circulating tumor cells (CTCs), such as B16F10 melanoma. This method provided, for the first time, identification of rolling CTCs in analogy to rolling white blood cells and CTC aggregates. Specifically, monitoring of PA signal widths from CTCs in mouse ear microvessels revealed double maxima in peak-width histograms associated with the fast moving portion of CTCs in central flow and slowly rolling CTCs in analogy to white blood cells. We also developed a two-parameter plot representing PA peak amplitude and peak widths. This method allowed identification of fast-moving individual CTCs, CTC aggregates, and rolling CTCs. The discovery of rolling CTCs in relatively large blood vessels indicates a higher probability of CTC extravasations, further increasing the possibility of metastasis through rolling mechanism in addition to mechanical capturing of CTCs in small vessels.