Two-photon microscopy enables in vivo imaging of astrocytic Ca2+ activity, yet detecting weak, transient, and background-coupled signals remains challenging due to low signal-to-noise ratios and heterogeneous noise. Here, we propose a low-parameter, adaptive framework for detecting weak astrocytic Ca2+ signals in two-photon imaging. After short-window frame accumulation, static background suppression, and Gaussian smoothing to stabilize intensity statistics, signal candidates are identified via segment-wise Gaussian mixture modeling, temporal persistence masking, and adaptive threshold updates. In simulated videos, the proposed method improved the Dice coefficient from 0.06 to 0.77 and increased the reference SNR from −9.82 to 3.40 dB. In in vivo recordings, the local SNR increased from 5.58 to 7.28 dB. Compared with fixed thresholding, AQuA, and AQuA2, our method was more robust under high-noise conditions while requiring only three user-defined parameters (minimum area, minimum duration, and an initialization coefficient). This framework provides an interpretable and computationally practical front-end module for the robust extraction of astrocytic Ca2+ signal in low-SNR two-photon imaging.
Xu et al. (Thu,) studied this question.