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This invention discloses an intracellular imaging system for simultaneously detecting UDG and miR-375 expression levels and its applications. The intracellular imaging system uses a tetrahedral DNA nanostructure as a carrier, loading a HAT-UDG probe with an on / off conformation, and coupling it with a split CRISPR / Cas12a detection component for recognizing miR-375. The split CRISPR / Cas12a detection component includes at least Cas12a protein, scaffold RNA, spacer RNA / equivalent split crRNA components, and a fluorescent reporter substrate. This invention is the first to perform a logical AND operation between two key indicators: UDG enzyme activity and miR-375 expression level. The system can only ultimately trigger a complete fluorescence signal when both are simultaneously highly expressed. This multi-target synergistic activation design constructs a precise molecular recognition logic gate, enabling extremely accurate differentiation between prostate cancer cells (RV-1) and normal cells (293T), as well as other cancer cells (5637) that highly express UDG, fundamentally avoiding misjudgments caused by fluctuations in a single indicator or non-specific expression, resulting in extremely high imaging specificity.