Digital PCR (dPCR) has transformed nucleic acid diagnostics by enabling the absolute quantification of rare mutations and target sequences. However, traditional dPCR detection methods, such as those involving flow cytometry and fluorescence imaging, may face challenges due to high costs, complexity, limited accuracy, and slow processing speeds. In this study, SAM-dPCR is introduced, a training-free open-source bioanalysis paradigm that offers swift and precise absolute quantification of biological samples. SAM-dPCR leverages the robustness of the zero-shot Segment Anything Model (SAM) to achieve rapid processing times (<
4 seconds) with an accuracy exceeding 97.10%. This method has been extensively validated across diverse samples and reactor morphologies, demonstrating its broad applicability. Utilizing standard laboratory fluorescence microscopes, SAM-dPCR can measure nucleic acid template concentrations ranging from 0.154 copies µL