Two-dimensional (2D) light scattering microscopy is one of the label-free imaging technologies that allows to image the particles out of focal plane. In this manuscript, we try to develop a volume scanning-based 2D light scattering microscopy that aims to fast determine the 3D location of single particles by taking the advantage of defocusing strategy, which is expected to improve the efficiency of common refocusing-based localization methods. To demonstrate the principle of our volume scanning-based 2D light scattering microscopy, the standard beads are used to perform the experiments. The vertical position of individual beads can be determined by measuring the area of the scattering patterns. The volume scanning-based 2D light scattering microscopy is proposed to provide a fast and label-free method for 3D localization of single particles.
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