arXiv:2505.18183v2 Announce Type: replace Abstract: Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disorder characterized by motor neuron degeneration, with changes to neuronal excitability being an early phenotype for ALS identified in patients and animal models. Recent advancements in induced pluripotent stem cell (iPSC) technology have enabled the generation of human iPSC-derived neuronal cultures, which, when combined with multi-electrode array (MEA) electrophysiology, provide rich spatial and temporal electrophysiological data. Traditionally, MEA data is analyzed using handcrafted features based on potentially imperfect domain knowledge, which while useful may not fully capture all the relevant characteristics inherent in the MEA data. Machine learning, in particular deep learning has the potential to automatically learn relevant characteristics (features) from raw data, without solely relying on handcrafted feature extraction. However, handcrafted features remain critical for encoding domain knowledge and improving model interpretability, especially in scenarios with limited or noisy data, as is often the case in most experimental studies. This study introduces FRAME-C, a knowledge-augmented machine learning pipeline that combines domain knowledge, raw spike waveform data, and deep learning techniques to classify MEA signals and identify ALS-specific phenotypes. FRAME-C leverages deep learning to extract important features from spike waveforms, while also incorporating handcrafted features such as spike amplitude, inter-spike interval, and spike duration, thus preserving key spatial and temporal information. We validated FRAME-C on both simulated and real-world MEA data from human iPSC-derived neuronal cultures, demonstrating its superior performance compared to existing methods for MEA classification.