Objective: To compare the fluorescence intensity and duration of qdots streptavidin conjugate (QDs-SA) with Cy3 as the molecular probe of β-amyloid precursor protein (APP), and to provide evidence for early molecular imaging and diagnosis of Alzheimer's disease (AD).
Methods: With the help of laser scanning confocal microscope and flow cytometry, the flurescence probe based on the QDs-SA was used to detect APP in HEK293 cells stably transfected pcDNA3.1/APP, and to compare with conventional fluroimmunoassay Cy3.
Results: The immunofluorescence staining detection indicated APP expression was mainly located in the plasma membrane. The mean fluorescence intensity of QDs-SA (34.2336±4.2455) was greater than that of Cy3 (21.6023±3.0102)under the confocal fluorescence microscope (P<0.05). After persistent exciting for 12 min, the fluorescence intensity of APP stained by QDs-SA decreased by 27.87%. The other stained by Cy3 decreased by 79.60%. The positive rate of APP staining had no significant difference between the QDs-SA(54.4700±3.4433)% and Cy3 (54.3800±8.5229)% by flow cytometry, but the mean fluorescence intensity had statistical significance(P<0.05). The QDs-SA (1 045.4167±47.3623) was significantly higher than the mean fluorescence intensity of Cy3 (658.5467±55.0591).
Conclusion: QDs-SA fluorescence probes can effectively recognize APP and are sensitive and exceptionally photostable, suggesting that QDs-SA fluorescence probes could be a potential method in APP detection and offer a novel way for the diagnosis of Alzheimer's disease.