Standardized Protocol of Cortical Neuronal Morphology Analysis Using High-Content Screening Systems

초록

Live-cell imaging is an indispensable tool for real-time observation of neuronal dynamics, critically enabling mechanistic studies of neurodevelopmental disorders. This study details the comprehensive characterization and optimization of a high-content screening (HCS) imaging system specifically tailored for morphological and functional neurobiological assays. Our HCS system is engineered for high-throughput, real-time visualization of intracellular signaling and dynamic molecular mechanisms in living cells. It is equipped with a high speed spinning disk confocal module and a highly sensitive scientific sCMOS camera, providing superior temporal and spatial resolution while effectively minimizing phototoxicity. The system accommodates a broad range of magnifications (4× to 60×), allowing for both high-resolution subcellular analysis and real-time monitoring of organelle behavior. Especially, the platform integrates specialized application modules, such as neurite outgrowth and cell cycle progression analysis. Its capacity for simultaneous long-term imaging of up to 96 individual samples positions it as a highly efficient solution for high throughput screening of cellular phenotypes. Complementary software facilitates automated, rapid image acquisition and streamlined data processing. The Brain Research Core Facilities at the Korea Brain Research Institute (KBRI) offer comprehensive support for advanced imaging modalities, including analyses of neuronal morphology. our objective is to provide standardized, high-throughput morphological imaging protocols to support the mechanistic dissection of brain function and elucidate cellular and structural correlates underlying diverse neuropathological conditions.