Developing highly sensitive and highly selective assays for monitoring heparin levels in blood is required during and after surgery. In previous studies, electrostatic interactions are exploited to recognize heparin and changes in light signal intensity are used to sense heparin. In the present study, we developed a quantum dot (QD) aggregation-based detection strategy to quantify heparin. When cationic micelles and fluorescence QDs modified with anti-thrombin III (AT III) are added into heparin sample solution, the AT III-QDs, which specifically bind with heparin, aggregate around the micelles. The aggregated QDs are recorded by spectral imaging fluorescence microscopy and differentiated from single QDs based on the asynchronous process of blue shift and photobleaching. The ratio of aggregated QD spots to all counted QD spots is linearly related to the amount of heparin in the range of 4.65 × 10 -4 U/mL to 0.023 U/mL. The limit of detection is 9.3 × 10 -5 U/mL (∼0.1 nM), and the recovery of the spiked heparin at 0.00465 U/mL (∼5 nM) in 0.1% human plasma is acceptable.
Keywords: aggregation; anti-thrombin III; heparin; quantum dot; single particle; spectral imaging microscopy.