Stars2Cells: Astrometric Tracking of Neurons Across Imaging Sessions

Chronic calcium imaging offers a window into how single neurons and ensemble activity change across days where identifying the same neurons from one session to the next is the prerequisite for answering questions regarding learning, drift, and plasticity over time. Yet only [~]2-3% of imaging laboratories publish longitudinal cross-session work, because existing registration tools depend on spatial-footprint or temporal correlations that degrade under repeated recording sessions. Here, we introduce Stars2Cells (S2C), a tracking pipeline inspired by astrometric plate-solving that represents each neurons local geometry as a four-dimensional quad descriptor invariant to rotation, translation, and uniform scaling. S2C operates purely on centroid coordinates and combines descriptor-space matching, Random Sample Consensus (RANSAC) verification, and Hungarian assignment. Across a synthetic benchmark of 1,262 paired runs spanning 100-1,000 neurons and 8 perturbation conditions plus 1 identity sanity-floor, S2C reached pooled F1 = 98.4% compared to the standard ROI-based matching of 36.0%. To show what this enables, we applied the pipeline to dorsomedial striatum (DMS) imaging during oral fentanyl behavioral-economics self-administration. Here, we show that a conserved population-rewarded lever press response in DMS masks near-complete single-neuron turnover. This representational-drift signature we demonstrated is invisible to the bulk photometry, and resolving it requires the same-cell tracking S2C provides. S2C is distributed as a GUI-driven standalone application for both macOS and Windows, requiring no Python, command line, or virtual environment setup.