Surface chemistry and deposition techniques
First Claim
1. A method for single molecule visualization, the method comprising:
- providing a surface, wherein the surface is a glass surface further comprising a uniform metal film, and wherein the surface is produced by a method comprising depositing a plurality of surface attachment features for specifically immobilizing fluorophore-labeled molecules on said surface by a deposition method that actively forces individual fluorophore-labeled molecules to be spaced apart by a distance equal to at least the diffraction limit for said fluorophore, wherein the deposition method provides at least a 3-fold increase in the maximum number of resolvable fluorophore-labeled molecules, as compared to random deposition;
immobilizing a plurality of fluorophore-labeled molecules via the plurality of surface attachment features; and
performing real time imaging for single fluorophores using surface plasmon resonance (SPR) fluorescence microscopy.
3 Assignments
0 Petitions
Accused Products
Abstract
Surface chemistries for the visualization of labeled single molecules (analytes) with improved signal-to-noise properties are provided. To be observed, analyte molecules are bound to surface attachment features that are spaced apart on the surface such that when the analytes are labeled adjacent analytes are optically resolvable from each other. One way to express this concept is that binding elements should be spaced apart such that the Guassian point spread functions of adjacent labels do not overlap. Another way of expressing this concept is that the surface binding elements should be spaced apart by a distance equal to at least the diffraction limit for an optical label attached to the bound analytes.
18 Citations
16 Claims
-
1. A method for single molecule visualization, the method comprising:
-
providing a surface, wherein the surface is a glass surface further comprising a uniform metal film, and wherein the surface is produced by a method comprising depositing a plurality of surface attachment features for specifically immobilizing fluorophore-labeled molecules on said surface by a deposition method that actively forces individual fluorophore-labeled molecules to be spaced apart by a distance equal to at least the diffraction limit for said fluorophore, wherein the deposition method provides at least a 3-fold increase in the maximum number of resolvable fluorophore-labeled molecules, as compared to random deposition; immobilizing a plurality of fluorophore-labeled molecules via the plurality of surface attachment features; and performing real time imaging for single fluorophores using surface plasmon resonance (SPR) fluorescence microscopy. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16)
-
Specification