Real-Time Imaging and Optimization of Cortical Neuron Morphology Analysis Using High-Content Screening Imaging Systems

초록

Live-cell imaging provides a powerful platform for real-time observation of neuronal dynamics, thereby facilitating mechanistic studies of neurodevelopmental disorders. In the present study, we conducted a detailed characterization of morphological dynamics and optimization strategies associated with high-content screening (HCS) imaging systems. Currently, HCS imaging systems are instrumental in enabling real-time visualization of intracellular signaling pathways and dynamic molecular mechanisms in living cells. Our system is equipped with a high-speed spinning disk confocal module and a highly sensitive scientific sCMOS camera, which together offer superior temporal and spatial resolution while minimizing phototoxic effects on live specimens. The imaging system accommodates a broad range of magnifications (4× to 60×), permitting high-resolution analysis of subcellular structures and real-time monitoring of dynamic organelle behavior. Furthermore, the platform integrates specialized application modules—such as neurite outgrowth and cell cycle progression analysis—which enhance its utility in targeted neurobiological and cell-based assays. The capacity for simultaneous long-term imaging of up to 96 individual samples positions the system as a highly efficient solution for high-throughput screening of morphological and functional cellular phenotypes. Complementary software tools provide an intuitive interface that supports automated, rapid image acquisition and streamlined data processing workflows. The Brain Research Core Facilities at the Korea Brain Research Institute (KBRI) offer comprehensive support for advanced imaging modalities, including analyses of neuronal morphogenesis and ultrastructural mapping. Our overarching objective is to refine and standardize morphological imaging protocols to support the mechanistic dissection of brain function and elucidate cellular and structural correlates underlying diverse neuropathological conditions.