Fluorescence resonance energy transfer (FRET) between two quantum dots of different sizes causes fluorescence quenching. Hereby a binding site pre-blocking approach is proposed to avoid this effect. Pre-binding of glucose on the donor occupies the binding sites and thus blocks resonance energy transfer between the two quantum dots, protecting the fluorescence from being quenched. A glucose assay is developed based on this approach. The glucose content is correlated with the fluorescence difference in the absence and in the presence of glucose. In practice, Green QDs-Con A conjugates are used as donors and Red QDs-NH(2)-glu conjugates as acceptors to form FRET system. The inhibition of fluorescence quenching is then measured in the presence of glucose. A linear calibration graph is achieved within 0.1-2.0 mmolL(-1), along with a detection limit of 0.03 mmolL(-1) and a RSD of 2.1% (1.0 mmolL(-1)). 91-105% of glucose in serum and urine samples is recovered. It is worth mentioning that the present glucose assay approach also generates a fluorescence chromatic difference imaging, and the color display clearly identifies the glucose contents by visual detection with a distinguishing ability of ca. 0.5 mmolL(-1). The present approach can potentially be used for the clinical determination of glucose in biological samples which can be further developed into a glucose sensor.
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http://dx.doi.org/10.1016/j.bios.2011.11.031 | DOI Listing |
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