Alzheimer's disease is a fatal, incurable, chronic neurodegenerative disease. Diagnosis in its early and even preclinical stages will be beneficial for its prevention and treatment. In the accepted pathological theory, abnormal accumulation of Aβ protein and abnormal mitochondrial function, including changes in mitochondrial viscosity, is closely related to Alzheimer's disease. To date, rare fluorescent probes have been reported that can simultaneously image Aβ plaques and mitochondrial viscosity. Therefore, the development of a dual-functional fluorescent probe for real-time fluorescence imaging of Aβ plaques and mitochondrial viscosity is crucial to discover a novel approach and strategy for the treatment of Alzheimer's disease, and to understand the pathological process and crosstalk between different biomarkers of Alzheimer's disease. Herein, we rationally designed and synthesized a series of fluorescent probes QM-SF-1∼5 with dimethylamino-quinolinium as the skeleton and thiophene as the π bridge to connect the groups with different electron-push/pull capacities. Among them, QM-SF-2 exhibited excellent properties such as large Stokes shift (168 nm), near-infrared emission (689 nm), and high selectivity and sensitivity (limit of detection was 1.07 μM) to Aβ aggregate and mitochondrial viscosity changes, indicating its promising prospects as a dual-functional imaging tool in the pathological study of Alzheimer's disease.Copyright © 2023 Elsevier B.V. All rights reserved.