Wang, J.; Russell, C.; Walczak, M.; Gao, D.; Qiao, G.; Fan, X.; Chu, C.; Janowski, M.; Walczak, P.; Liang, Y.

Intracerebral cell transplantation holds great promise for treating stroke and other neurological disorders, but progress is hindered by challenges in precise cell delivery and limited methods for monitoring post-engraftment dynamics. To address these limitations, we introduce OPTRACE (OPtical imaging-guided Transplantation and tRAcking of CElls), a two-step framework that integrates precision delivery with longitudinal in vivo tracking to advance cell therapy. During transplantation, OPTRACE enables real-time visualization using cost-effective translucent glass micropipettes, guided by predictive modeling to optimize delivery parameters. A novel pulse-elevation injection technique further enhances the precision of superficial cortical engraftment. Following transplantation, multicolor labeling combined with two-photon microscopy permits longitudinal single-cell tracking, revealing host microglial responses, and altered neuronal calcium signaling at the graft interface. By unifying high-resolution delivery and tracking strategies, OPTRACE provides a powerful platform for elucidating engraftment biology and accelerating the development of effective cell-based interventions for neurological diseases.