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Non‐invasive screening for bladder cancer is crucial for treatment and postoperative follow‐up. This study combines digital microfluidics (DMF) technology with fluorescence lifetime imaging microscopy (FLIM) for urine analysis and introduces a novel non‐invasive bladder cancer screening technique. Initially, the DMF was utilized to perform preliminary screening and enrichment of urine exfoliated cells from 54 participants, followed by cell staining and FLIM analysis to assess the viscosity of the intracellular microenvironment. Subsequently, a deep learning residual convolutional neural network was employed to automatically classify FLIM images, achieving a three‐class prediction of high‐risk (malignant), low‐risk (benign), and minimal risk (normal) categories. The results demonstrated a high consistency with pathological diagnosis, with an accuracy of 91% and a precision of 93%. Notably, the method is sensitive for both high‐grade and low‐grade bladder cancer cases. This highly accurate non‐invasive screening method presents a promising approach for bladder cancer screening with significant clinical application potential. [ABSTRACT FROM AUTHOR]