Three Dimensional Lifetime-Multiplex Tomography Based on Time-Gated Capturing of Near-Infrared Fluorescence Images

We report a computed tomography (CT) technique for mapping near-infrared fluorescence (NIRF) lifetime as a multiplex three-dimensional (3D) imaging method, using a conventional NIR camera. This method is achieved by using a time-gated system composed of a pulsed laser and an NIR camera synchronized with a rotatable sample stage for NIRF-CT imaging. The fluorescence lifetimes in microsecond-order of lanthanides were mapped on reconstructed cross-sectional and 3D images, via back-projection of two-dimensional projected images acquired from multiple angles at each time point showing fluorescence decay. A method to select slopes (the observed decay rates in time-gated imaging) used for the lifetime calculation, termed as the slope comparison method, was developed for the accurate calculation of each pixel, resulting in reduction of image acquisition time. Time-gated NIRF-CT provides a novel choice for multiplex 3D observation of deep tissues in biology.