This study integrates Near Infrared Spectroscopy (NIRS) and nanoscale imaging technologies to discern alterations in muscle tissue biomarkers, thereby enhancing the precision of non-invasive monitoring of muscle fatigue. Experimental investigations were carried out on the biceps brachii
muscle of 12 subjects, categorized into mild, moderate, and severe fatigue groups. Concurrently, a specific wavelength of Near Infrared Laser Diode (NIR-LD) was employed to acquire spectral data. The application of Atomic Force Microscopy (AFM) in conjunction with NIRS imaging facilitated
the attainment of high-resolution images of the biceps brachii tissue. The absorption characteristics of distinct biomarkers in muscle tissue, responsive to near-infrared light, were captured to calculate concentration variations and evaluate muscle fatigue levels. The findings revealed substantial
variations in the concentrations of Oxy-hemoglobin (HbO), Deoxy-hemoglobin (HbR), Lactic Acid (LA), Phosphocreatine (PCr), Troponin (Tn), Creatine Kinase (CK), and Glutamine (Gln) across different fatigue groups. Muscle fatigue assessment exhibited an average sensitivity, accuracy, specificity,
and F1 score of 0.96, 0.96, 0.95, and 0.96, respectively, for the 12 subjects. The average Area Under the Curve (AUC) values for detecting mild, moderate, and severe fatigue were 0.96, 0.96, and 0.98, respectively. This method demonstrates notable accuracy in the identification of muscle fatigue
biomarkers, rendering it suitable for sports-related muscle fatigue assessment.