Fluorescence immunoassay device based on integration of photonic crystal and magnetic beads
A fluorescence immunoassay device based on integration of a photonic crystal and magnetic beads and a method thereof are provided. Magnetic beads with high surface-to-volume ratio are used as carriers of fluorescent molecules to obtain higher fluorescence density. The electric field on the surface of the photonic crystal is enhanced through excitation of photonic crystal resonance. The intensity of the fluorescence signal excited by the enhanced electric field is increased. Moreover, through interaction with the photonic crystal, some fluorescent signals that originally cannot be received by the fluorescent sensor are coupled to the photonic crystal resonant modes and reradiate toward the fluorescent sensor, thereby increasing collection efficiency. The fluorescence signals generated by fluorescent molecules on the magnetic beads are significantly intensified, which could lower the detection limit. Furthermore, the magnetic beads aggregation method can achieve the detection capability that cannot be achieved by the current fluorescent immunoassay.
1 . A fluorescence immunoassay device based on integration of a photonic crystal and magnetic beads, comprising
a photonic crystal, said photonic crystal being a resonant waveguide grating structure;
at least one magnetic bead disposed on a surface of said photonic crystal that is specific for a tested biological molecule;
a plurality of the fluorescent molecules that are specific for the tested biological molecule;
a light source providing an incident exciting light, wherein said incident exciting light resonates with said photonic crystal to enhance an electric field on said surface of said photonic crystal; said electric field further excites said fluorescent molecules to generate fluorescence signals intensified by said electric field; and
a florescence detector receiving said fluorescence signals that said fluorescent molecules emit to said fluorescence detector and said fluorescence signals that are reflected from said photonic crystal, and processing said fluorescence signals into a sensed image.
2 . The fluorescence immunoassay device based on integration of a photonic crystal and magnetic beads according to claim 1 , wherein said tested biological molecules have a concentration of at least 10 −3 pg/ml.
3 . The fluorescence immunoassay device based on integration of a photonic crystal and magnetic beads according to claim 1 , wherein said tested biological molecule is selected from a group consisting of nucleic acids, antigens, antibodies, binding proteins, phytohematoagglutinin, hormone receptors, and small-molecule compounds.
4 . The fluorescence immunoassay device based on integration of a photonic crystal and magnetic beads according to claim 1 comprising a plurality of magnetic beads.
5 . The fluorescence immunoassay device based on integration of a photonic crystal and magnetic beads according to claim 4 further comprising a magnetic device, wherein said magnetic device neighbors said magnetic beads and attracts said magnetic beads to aggregate on said surface of said photonic crystal.
6 . The fluorescence immunoassay device based on integration of a photonic crystal and magnetic beads according to claim 5 , wherein said tested biological molecules have a concentration of at least 10 −4 pg/ml.
7 . The fluorescence immunoassay device based on integration of a photonic crystal and magnetic beads according to claim 1 , wherein said resonant waveguide grating structure includes a substrate, a grating layer and a waveguide layer; said grating layer is formed on said substrate; said waveguide layer is formed on said grating layer.
8 . The fluorescence immunoassay device based on integration of a photonic crystal and magnetic beads according to claim 1 , wherein said fluorescence detector detects a plurality of pixels of said sensed image and sets said pixels whose pixel values are smaller than a black balance value to be zero to eliminate noise of said sensed image.
9 . The fluorescence immunoassay device based on integration of a photonic crystal and magnetic beads according to claim 1 further comprising an image processor, wherein said image processor receives said sensed image and analyzes intensities of said fluorescence signals according to said sensed image.