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) in deep tissues. However, the broad application of this method is hindered by its long scanning time, poor accuracy, and low stability. This is mainly because most PALT systems execute the three data acquisition sequences separately without automatic control and neglect the long-time fluctuation of the laser output. In this work, we have proposed a novel automatic interleaved data acquisition method for PALT. This new method not only improved the scanning efficiency but also eliminated the long-time fluctuations of laser pulse energy. Results show that this new method can significantly improve the system's stability and help reduce the scanning time. With this new method, we obtained the 3D background-free TTD images for the first time. We also observed distinct hypoxia inside the tumor due to the high metabolic rate of cancer cells, demonstrating the high reliability of our proposed method. The proposed method in this work can significantly promote the application of PALT imaging in biomedical studies.
Wang et al. (Thu,) studied this question.