MODIFIED SURFACES FOR THE DETECTION OF BIOMOLECULES AT THE SINGLE MOLECULE LEVEL
First Claim
Patent Images
1. A surface composition comprising:
- a substrate and a functionalized layer disposed on a surface thereof, an absorption layer disposed on a surface thereof, or an absorption layer disposed on a functionalized layer disposed on a surface thereof, where the functionalized layer includes sites capable of the absorption layer or capable of absorbing or binding a first molecule, molecular complex, or molecular assembly or a plurality of first molecules, complexes, assemblies or mixtures or combinations thereof where the absorption layer include sites capable of absorbing or binding the first molecule, molecular complex, or molecular assembly or the plurality of first molecules, complexes, assemblies or mixtures or combinations thereof, where each first molecule, complex and assembly includes a first fluorescent dye or fluorophore, and where the composition increases fluorescent dye or fluorophore lifetime relative to a polyelectrolyte surface.
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Abstract
Support surfaces are disclosed that are designed to support molecules or molecular assemblies immobilized thereon so that the molecules or molecular assemblies can be observed in single molecule detections systems, where the support surfaces have reduced background and the fluorescent labels associated with the immobilized molecules or molecular assemblies have longer active lifetimes prior to permanent photo-bleaching or deactivation and have improve fluorescence properties and where the surfaces have more uniform fluorescent properties.
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Citations
30 Claims
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1. A surface composition comprising:
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a substrate and a functionalized layer disposed on a surface thereof, an absorption layer disposed on a surface thereof, or an absorption layer disposed on a functionalized layer disposed on a surface thereof, where the functionalized layer includes sites capable of the absorption layer or capable of absorbing or binding a first molecule, molecular complex, or molecular assembly or a plurality of first molecules, complexes, assemblies or mixtures or combinations thereof where the absorption layer include sites capable of absorbing or binding the first molecule, molecular complex, or molecular assembly or the plurality of first molecules, complexes, assemblies or mixtures or combinations thereof, where each first molecule, complex and assembly includes a first fluorescent dye or fluorophore, and where the composition increases fluorescent dye or fluorophore lifetime relative to a polyelectrolyte surface. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11)
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12. A surface composition comprising:
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a substrate, a functionalized layer disposed on a surface thereof, and an absorption layer disposed on the functionalized layer, where the functionalized layer is adapted to bond or absorb the absorption layer, where the absorption layer include sites capable of absorbing or binding a first molecule, molecular complex, or molecular assembly or a plurality of first molecules, complexes, assemblies or mixtures or combinations thereof, where each first molecule, complex and assembly includes a first fluorescent dye or fluorophore, and where the composition increases fluorescent dye or fluorophore lifetime relative to a polyelectrolyte surface. - View Dependent Claims (13, 14, 15, 16, 17, 18, 19, 20, 21, 22)
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23. A method for preparing surface compositions comprising the steps of:
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cleaning a substrate to reduce or eliminate fluorophores from surfaces of the substrate or from the substrate itself, and contacting the substrate with an absorbent under conditions to form an absorption layer on a surface of the substrate, where the absorption layer include sites capable of absorbing or binding a first species selected from the group consisting of a molecule, molecular complex, or molecular assembly or a plurality of first species selected from the group consisting of molecules, complexes, assemblies or mixtures or combinations thereof, where each first species includes a first fluorescent dye or fluorophore, and where the substrate increases fluorescent dye or fluorophore lifetime relative to a polyelectrolyte surface. - View Dependent Claims (24)
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25. A method for preparing surface compositions comprising the steps of:
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2 cleaning a substrate to reduce or eliminate fluorophores from surfaces of the substrate or from the substrate itself, placing the cleaned substrate on a support rack in a bottle including a cap having an aperture and a bottom having a plurality of aperture, placing a tube within the bottle containing a functionalizing agent, capping the bottle, inserting a pipette through the cap aperture into the tube above or below a surface of the functionalizing agent, connecting the pipette to a source of an inert gas, supplying a flow of the inert gas to the bottle via the pipette at a rate sufficient to evaporate or entrain the functionalizing agent in the inert gas flow and sufficient for a pressure in the bottle to be maintained at a desired pressure below the rupture pressure of the bottle, and continuing the flow for a time sufficient to achieve a desired degree of substrate functionalization to form a substrate having a functionalized layer formed one or all surfaces of the substrate, where the functionalized layer is adapted to bond, absorb or support the absorption layer, or to absorb or bond a first species selected from the group consisting of a molecule, molecular complex, or molecular assembly or a plurality of first species selected from the group consisting of molecules, complexes, assemblies or mixtures or combinations thereof, where each first species includes a first fluorescent dye or fluorophore, and where the composition increases fluorescent dye or fluorophore lifetime relative to a polyelectrolyte surface.
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26. A method for immobilizing molecules, molecular complexes or molecular assemblies comprising the steps of:
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providing a substrate comprising an absorption layer disposed on a surface thereof, an absorption layer disposed on a functionalized layer disposed on a surface thereof, or a functionalized layer disposed on a surface thereof, contacting the surface with a solution comprising a plurality of species selected from the group consisting of molecules, molecular complexes, molecular assemblies or mixtures or combinations thereof, where some or all of the species include a first detectable label, where the contacting is under conditions sufficient to immobilize the species on the functionalized layer or the absorption layer so that a majority of the immobilized species are separately or individually detectable, placing the resulting composition in a detection system, and detecting the labels. - View Dependent Claims (27, 28, 29, 30)
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Specification