Mixed-metal substrates for metal-enhanced fluorescence
The present invention provides for mixed metal structures that can be deposited on a substrate or free in solution that exhibit several distinctive properties including a broad wavelength range for enhancing fluorescence signatures. Further, metal surface plasmons can couple and such diphase coupled luminescence signatures create extra plasmon absorption bands. The extra bands allow for a broad range of fluorophores to couple therefore making more generic substrates with wider reaching applications.
1. A detection system, the system comprising:
a multiplicity of metallic structures, wherein the metallic structures are metallic particles having a geometric shape selected from the group consisting of a sphere, triangle, square, oblong, elliptical and rectangle and wherein the metallic particles are separated from each other at a distance of 40 to 50 nm, wherein each of the metallic particles comprises mixed-metals and are fabricated from a combination of two plasmon supporting metals wherein the two plasmon supporting metals comprise a silver metallic particle having a coating from about 2 nm to about 8 nm of aluminum deposited thereon, wherein the metallic structures further comprise a capture probe having affinity for a target molecule in a sample used in the detection system, wherein the two plasmon supporting metals provide an additional plasmon resonance band not present in the plasmon absorption spectra of either silver or aluminum, wherein the metallic structures are immobilized on a substrate and the substrate is selected from the group consisting of glass, quartz, polymeric materials, and cellulose;
at least one excitable molecule that is positioned near the multiplicity of metallic structures in a range from about 5 nm to 30 nm from the multiplicity of metallic structures, wherein the excitable molecule is selected from the group of an intrinsic fluorophore, extrinsic fluorophore, fluorescent dye, a luminophore, a chemiluminescent species and a bioluminescent species and wherein the excitable molecule is attached to a free probe and wherein the binding of the free probe to the target molecule causes the excitable molecule to be positioned from about 5 nm to 30 nm from the multiplicity of metallic structures;
a source of electromagnetic energy for exciting the excitable molecule if the excitable molecule is an intrinsic fluorophore, extrinsic fluorophore, fluorescent dye or a luminophore; and
a detector for detecting emissions from the excited molecule and/or the metallic structures.
2. The detection system according to claim 1 , wherein the intrinsic fluorophore is a protein.
3. The detection system according to claim 1 , wherein the excitable molecule emits a detectable signal upon excitation and when positioned from 5 nm to 20 nm from the metallic structures.
4. The detection system according to claim 1 , wherein the excitable molecule comprises a first and second component of a bioluminescence or chemiluminescence generating system.
5. A method for detecting emissions from an excitable molecule in a detection system, the method comprising:
providing a substrate comprising a multiplicity of metallic structures, wherein the metallic structures are metallic particles having a geometric shape selected from the group consisting of a sphere, triangle, square, oblong, elliptical and rectangle and wherein the metallic particles are separated from each other at a distance of 40 to 50 nm wherein each of the metallic particles comprises mixed-metals, wherein the metallic structures are fabricated from a combination of two plasmon supporting metals wherein the two plasmon supporting metals consist of a silver metallic particle having a coating from about 2 nm to about 8 nm of aluminum deposited thereon, wherein the metallic structures have positioned thereon a receptor molecule having affinity for a ligand of interest in a sample, wherein the ligand of interest binds to the receptor molecule to form a receptor-ligand complex and the excitable molecule binds to the receptor-ligand complex, wherein the at least two plasmon supporting metals provide an additional plasmon resonance band not present in the plasmon absorption spectra of either silver or aluminum, wherein the metallic structures are immobilized on a substrate and the substrate is selected from the group consisting of glass, quartz, polymeric materials, and cellulose;
positioning the excitable molecule near the multiplicity of metallic structures in a range from about 5 nm to 30 nm from the multiplicity of metallic structures, wherein the excitable molecule is selected from the group of an intrinsic fluorophore, extrinsic fluorophore, fluorescent dye, a luminophore, a chemiluminescent species and a bioluminescent species;
applying electromagnetic energy for exciting the excitable molecule if the excitable molecule is an intrinsic fluorophore, extrinsic fluorophore, fluorescent dye or a luminophore; and
detecting emissions from the excited molecule and/or the metallic structures.