This study developed an automated microfluidic platform capable of supporting long‐term culture, drug testing, and real‐time biomarker monitoring of PDOs, offering a method that enabled continuously detecting of biomarkers. By integrating miniaturized culture chambers with in situ biosensing arrays, the system enables continuous, non‐invasive monitoring of tumor biomarkers (e.g., CEA) at single‐cluster resolution (40∼50 organoids) over multiple time points. Here, CEACAM5 is linked to neoplasm.