The temporal focusing two-photon microscope (TFM) is developed to perform depth resolved wide field fluorescence imaging by capturing frames sequentially. However, due to strong nonignorable noises and diffraction rings surrounding particles, further researches are extremely formidable without a precise particle localization technique. In this paper, we developed a fully-automated scheme to locate particles positions with high noise tolerance. Our scheme includes the following procedures: noise reduction using a hybrid Kalman filter method, particle segmentation based on a multiscale kernel graph cuts global and local segmentation algorithm, and a kinematic estimation based particle tracking method. Both isolated and partial-overlapped particles can be accurately identified with removal of unrelated pixels. Based on our quantitative analysis, 96.22% isolated particles and 84.19% partial-overlapped particles were successfully detected.
Xia Huang, Chunqiang Li, Chuan Xiao, Wenqing Sun, and Wei Qian, "A fully-automated multiscale kernel graph cuts based particle localization scheme for temporal focusing two-photon microscopy," Proc. SPIE 10137, Medical Imaging 2017: Biomedical Applications in Molecular, Structural, and Functional Imaging, 101371I (Presented at SPIE Medical Imaging: February 15, 2017; Published: 13 March 2017); https://doi.org/10.1117/12.2254567.
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