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The diagnosis of Amyotrophic Lateral Sclerosis (ALS) primarily relies on clinical findings and remains challenging, particularly in the early stages of the disease, where characteristic symptoms may be subtle and nonspecific. Therefore, the development of disease-specific and clinically validated biomarkers is crucial to enhance diagnostic precision and optimize patient management. Here, we explored tear fluid (TF) as a promising biomarker source for ALS, given its accessibility, prior evidence ...