PREPARATION OF ARRAY SURFACES FOR SINGLE-ANALYTE PROCESSES
Compositions and methods are provided for forming single-analyte arrays with enhanced characteristics for inhibiting orthogonal binding of molecules to the array. Arrays are modified to contain covalently incorporated passivating moieties at array addresses where orthogonal binding may occur. The compositions and methods may facilitate detection of an increased quantity of array features at single-analyte resolution.
1 ) A method, comprising:
a) providing a substrate, wherein the substrate comprises a solid support and a layer of a resist material disposed on a surface of the solid support, and wherein the layer of the resist material comprises a plurality of depressions, wherein each individual depression of the plurality of depression comprises an exposed region of the surface of the solid support;
b) binding a plurality of molecules to each individual exposed region of the surface of the solid support in each individual depression of the plurality of depressions, wherein molecules of the plurality of molecules comprise reactive functional groups that are not attached to the surface of the solid support, wherein the binding occurs in the presence of a fluidic medium comprising water and a miscible solvent; and
c) after binding the plurality of molecules to each individual exposed region of the surface of the solid support, removing the layer of the resist material from the surface of the solid support.
2 ) The method of claim 1 , wherein a reactive functional group of the reactive functional groups comprises a nucleophilic functional group.
3 ) The method of claim 2 , wherein the nucleophilic functional group comprises an amine functional group.
4 ) The method of claim 2 , wherein a reactive functional group of the reactive functional groups is a terminal moiety of a molecule of the plurality of molecules.
5 ) The method of claim 1 , further comprising exposing the substrate to a plasma.
6 ) The method of claim 5 , wherein exposing the substrate to the plasma occurs for no more than 10 minutes.
7 ) The method of claim 5 , further comprising contacting the substrate with a reactive agent, wherein the contacting occurs after exposing the substrate to the plasma and before removing the layer of the resist material.
8 ) The method of claim 7 , wherein the contacting comprises contacting the substrate with a gas-phase reactive agent.
9 ) The method of claim 7 , wherein the contacting comprises contacting the substrate with a liquid-phase reactive agent.
10 ) The method of claim 1 , wherein the miscible solvent comprises an aprotic solvent.
11 ) The method of claim 10 , wherein the aprotic solvent is selected from the group consisting of N-methyl pyrrolidone, tetrahydrofuran, ethyl acetate, acetone, dimethylfuran, acetonitrile, dimethyl sulfoxide, propylene carbonate, and a combination thereof.
12 ) The method of claim 1 , wherein the miscible solvent comprises a polar solvent.
13 ) The method of claim 12 , wherein the polar solvent is selected from the group consisting of N-methyl pyrrolidine, tetrahydrofuran, ethyl acetate, acetone, dimethylfuran, acetonitrile, dimethyl sulfoxide, propylene carbonate, N-butanol, isopropyl alcohol, nitromethane, ethanol, methanol, acetic acid, and a combination thereof.
14 ) The method of claim 9 , wherein a weight ratio of the water to the miscible solvent in the fluidic medium is no more than 10:1.
15 ) The method of claim 1 , wherein removing the layer of the resist material from the surface of the solid support comprises contacting the resist material with a fluidic medium comprising a surfactant.
16 ) The method of claim 15 , wherein the surfactant comprises a non-ionic surfactant, a cationic surfactant, an anionic surfactant, a zwitterionic surfactant, an amphoteric surfactant, or a combination thereof.
17 ) The method of claim 15 , wherein the contacting of the resist material with a fluidic medium occurs in the presence of sonication.
18 ) The method of claim 17 , wherein the sonication occurs for at least 10 minutes.
19 ) An array, comprising:
a) a solid support comprising a surface;
b) a plurality of discrete regions on the surface of the solid support, wherein each discrete region comprises a plurality of molecules coupled to the surface of the solid support; and
c) one or more interstitial regions, wherein each individual discrete region of the plurality of discrete regions is separated from each other discrete region by an interstitial region of the one or more interstitial regions;
wherein the plurality of molecules comprises a plurality of passivating molecules and a plurality of coupling molecules, wherein a ratio of a quantity of the plurality of passivating molecules to a quantity of the plurality of coupling molecules is at least 2:1; and
wherein the one or more interstitial regions comprise a layer disposed on the surface of the solid support, wherein the layer comprises a hydrophobic material.
20 ) A flow cell, comprising:
a) a first solid support, wherein an array comprising a plurality of discrete regions is disposed on a surface of the first solid support, wherein each discrete region comprises a plurality of molecules coupled to the surface of the solid support; and
b) a second solid support;
wherein the first solid support is joined to the second solid support to form an enclosed void, wherein the array is disposed within the void, and wherein a surface of the second solid support within the void comprises a layer of a hydrophobic material.