Sophia Liu, David Z. He, Veronica Gomez-Godinez, Zachary Wan1, Chengbiao Wu, Linda Shi

Abstract: Femtosecond laser ablation creates precise microinjuries that reveal how cells respond to damage linked to degenerative disease. A Robotic Laser Microscope (Robolase) setup, combined with fluorescence imaging and quantitative phase imaging (QPI), was applied across six new area of studies: primary mouse cortical neurons, cardiomyocytes, rat pheochromocytoma (PC12), human embryonic kidney (HEK293), hippocampal (wild type and Huntington’s disease, HD), and retinal ganglion cells（RGCs）. In cortical neurons, axonal cuts under normal (3 mM) versus high (25 mM) glucose consistently led to post-ablation retraction, supporting evaluation of axonal robustness under diabetic conditions. In cardiomyocytes, patterned ablation that simulated a loss-of-neighbor border zone produced measurable changes in border-cell area over time. QPI detected power-dependent, real-time thickness shifts in HEK293 and PC12 cells after ablation, providing label-free readouts of subcellular mass redistribution. In hippocampal neurons, HD cultures showed larger axonal separation than wild type; nicotinamide reduced degeneration in a dose-dependent manner, and H₂O₂ modestly increased it. In RGCs, axonal injury triggered rapid axonal Ca²⁺ spikes followed by delayed, larger somatic peaks that decayed over minutes. These results position laser ablation as a practical platform for quantifying structural and signaling dynamics after controlled injury and for screening candidate modulators of degeneration.

Keywords: Femtosecond laser ablation, axonal degeneration, border zone, quantitative phase imaging, calcium imaging, Huntington’s disease, retinal ganglion cells

Date Published: November 25, 2025
DOI: 10.11159/jbeb.2025.011

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