We focus on the issue of limited number of samples in the single particle tracking (SPT) when trying to extract the diffusion anisotropy that originates from the particle asymmetry. We propose a novel evaluation technique of SPT making use of the relation of the consecutive three steps. More specifically, the trend of the angle comprised of the three positions and the displacements are plotted on a scatter diagram. The particle anisotropy dependence of the shape of the scatter diagram is examined through the data from the standard numerical model of anisotropic two-dimensional Brownian motion. Comparison with the existing method reveals the faster convergence in the evaluation. In particular, our proposed method realizes the detection of diffusion anisotropy under the conditions of not only less number of data but also larger time steps. This is of practical importance not only when the abundant data is hard to achieve but also when the rotational diffusion is fast compared to the frame rate of the camera equipment, which tends to be more common for smaller particles or molecules of interest.
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