FLOW CELLS WITH PASSIVATION COMPONENTS
An example flow cell includes a substrate including depressions separated by interstitial regions; a polymeric hydrogel positioned within each of the depressions; a primer set attached to the polymeric hydrogel; and one of: a passivation component attached to the interstitial regions, or a passivation component attached to the polymeric hydrogel, or respective passivation components attached to each of the interstitial regions and the polymeric hydrogel.
1 . A flow cell, comprising:
a substrate including depressions separated by interstitial regions;
a polymeric hydrogel positioned within each of the depressions;
a primer set attached to the polymeric hydrogel; and
one of: a passivation component attached to the interstitial regions, a passivation component the polymeric hydrogel, or respective passivation components attached to each of the interstitial regions and the polymeric hydrogel, wherein:
the passivation component is exclusively attached to the interstitial regions and includes i) a silane, siloxane, or alkoxy silane small molecule with an alkoxy, hydroxyl, carboxylic acid, or perfluorinated end group, or ii) a first linker selected from the group consisting of alkylene, poly(ethylene glycol), poly(meth)acrylic acid, polyacrylamide, poly(2-hydroxyl ethyl (meth)acrylate), and poly(2-hydroxypropyl (meth)acrylamide) and a first end group selected from the group consisting of phosphate, phosphonate, a zwitterionic end group and an amphoteric polymer end group; or
the passivation component is exclusively attached to the polymeric hydrogel and includes i) a second linker selected from the group consisting of alkylene, poly(ethylene glycol), poly(meth)acrylate, polyacrylamide, pentaerythritol, and combinations thereof, and a second end group selected from the group consisting of a poly(ethylene glycol) end group when the linker is poly(meth)acrylate, polyacrylamide, pentaerythritol, and combinations thereof, a zwitterionic end group, a hydroxyl end group, a carboxylic acid end group, a sulfonic acid end group, a positively chargeable end group, an alkoxy end group, an anionic polymer end group, and an amphoteric polymer end group, or ii) a dibenzocyclooctyne-functionalized passivation agent; or
the respective passivation components are attached to each of the interstitial regions and to the polymeric hydrogel, the respective passivation component attached to the interstitial regions includes i) a silane, siloxane, or alkoxy silane small molecule with an alkoxy, hydroxyl, carboxylic acid, or perfluorinated end group, or ii) a third linker selected from the group consisting of alkylene, poly(ethylene glycol), poly(meth)acrylic acid, polyacrylamide, poly(2-hydroxyl ethyl (meth)acrylate), and poly(2-hydroxypropyl (meth)acrylamide) and a third end group selected from the group consisting of a zwitterionic end group and an amphoteric polymer end group, and the respective passivation component attached to the polymeric hydrogel includes i) a fourth linker selected from the group consisting of alkylene, poly(ethylene glycol), poly(meth)acrylate, polyacrylamide, and pentaerythritol, and combinations thereof, and a fourth end group selected from the group consisting of a poly(ethylene glycol) end group when the linker is poly(meth)acrylate, polyacrylamide, or pentaerythritol, a zwitterionic end group, a hydroxyl end group, a carboxylic acid end group, a sulfonic acid end group, a positively chargeable end group, an alkoxy end group, an anionic polymer end group, and an amphoteric polymer end group, or ii) a dibenzocyclooctyne-functionalized passivation agent.
2 . The flow cell as defined in claim 1 , wherein:
the passivation component is exclusively attached to the polymeric hydrogel; and
the second linker further includes an alkyne-containing functional group or an alkene-containing functional group attached to the polymeric hydrogel.
3 . The flow cell as defined in claim 2 , wherein:
the polymeric hydrogel includes terminal azide groups; and
the passivation component is selected from the group consisting of dibenzocyclooctyne-poly(ethylene glycol)-carboxylic acid, norbornene-poly(ethylene glycol)-carboxylic acid, bicyclo[6.1.0]non-4-yne-poly(ethylene glycol)-carboxylic acid, dibenzocyclooctyne-poly(ethylene glycol)-amine, norbornene-poly(ethylene glycol)-amine, bicyclo[6.1.0]non-4-yne-poly(ethylene glycol)-amine, dibenzocyclooctyne-poly(ethylene glycol)-hydroxyl, norbornene-poly(ethylene glycol)-hydroxyl, bicyclo[6.1.0]non-4-yne-poly(ethylene glycol)-hydroxyl, dibenzocyclooctyne-poly(ethylene glycol)-sulfonic acid, norbornene-poly(ethylene glycol)-sulfonic acid, and bicyclo[6.1.0]non-4-yne-poly(ethylene glycol)-sulfonic acid.
4 . The flow cell as defined in claim 2 , wherein:
the polymeric hydrogel includes terminal azide groups;
the second linker is poly(ethylene glycol)-dibenzocyclooctyne, poly(ethylene glycol)-norbornene, bicyclo[6.1.0]non-4-yne-poly(ethylene glycol), or bicyclo[6.1.0]non-4-yne-poly(ethylene glycol) methyl ether;
the second end group is the zwitterionic end group; and
the zwitterionic end group is selected from the group consisting of phosphocholine, a sulfobetaine, and a carboxybetaine.
5 . The flow cell as defined in claim 2 , wherein:
the polymeric hydrogel includes terminal azide groups;
the second linker is poly(ethylene glycol)-dibenzocyclooctyne, poly(ethylene glycol)-norbornene, bicyclo[6.1.0]non-4-yne-poly(ethylene glycol), or bicyclo[6.1.0]non-4-yne-poly(ethylene glycol) methyl ether;
the second end group is the amphoteric polymer end group; and
the amphoteric polymer end group includes both a carboxylic acid functional group and an amine functional group.
6 . The flow cell as defined in claim 2 , wherein:
the polymeric hydrogel includes terminal azide groups;
the second linker is poly(ethylene glycol)-dibenzocyclooctyne, poly(ethylene glycol)-norbornene, bicyclo[6.1.0]non-4-yne-poly(ethylene glycol), or bicyclo[6.1.0]non-4-yne-poly(ethylene glycol) methyl ether;
the second end group is the anionic polymer end group; and
the anionic polymer end group is poly(meth) acrylic acid.
7 . The flow cell as defined in claim 1 , wherein:
the passivation component is exclusively attached to the polymeric hydrogel;
the passivation component includes the dibenzocyclooctyne-functionalized passivation agent; and
the dibenzocyclooctyne-functionalized passivation agent is selected from the group consisting of dibenzocyclooctyne-acid, dibenzocyclooctyne-amine, and dibenzocyclooctyne-sulfo-amine.
8 . The flow cell as defined in claim 1 , wherein:
the passivation component is exclusively attached to the interstitial regions;
the first linker is poly(ethylene glycol) and further includes a silane functional group attached to the interstitial regions;
the first end group is the zwitterionic end group; and
the zwitterionic end group is selected from the group consisting of phosphocholine, a sulfobetaine, and a carboxybetaine.
9 . The flow cell as defined in claim 1 , wherein:
the passivation component is exclusively attached to the interstitial regions;
the first linker is poly(ethylene glycol) and further includes a silane functional group attached to the interstitial regions;
the first end group is the amphoteric polymer end group; and
the amphoteric polymer end group includes both a carboxylic acid functional group and an amine functional group.
10 . The flow cell as defined in claim 1 , wherein:
the passivation component is exclusively attached to the interstitial regions; and
the passivation component is selected from the group consisting of (heptadecafluoro-1,1,2,2-tetrahydrodecyl) trimethoxysilane, 3-(heptafluoroisopropoxy)propyltrimethoxysilane, nonafluorohexyltrimethoxysilane, [perfluoro(polypropyleneoxy)]methoxypropyltrimethoxysilane, 1,3-bis(trifluoropropyl)-1,1,3,3-tetramethyldisilazane, and (tridecafluoro-1,1,2,2-tetrahydrooctyl)dimethylchlorosilane.
11 . The flow cell as defined in claim 1 , wherein
the passivation component is exclusively attached to the interstitial regions; and
the passivation component is selected from the group consisting of N,N-bis(2-hydroxyethyl)-3-aminopropyltriethoxysilane, bis(methoxyethyl)-3-trimethoxysilylpropylammoniumchloride, carboxyethylsilanetriol disodium salt, and 3-(trihydroxysilyl)-1-propanesulfonic acid.
12 . The flow cell as defined in claim 1 , wherein:
the respective passivation components are attached to each of the interstitial regions and the polymeric hydrogel;
the respective passivation component attached to the interstitial regions is selected from the group consisting of 2-[methoxypoly(ethylenoxy)6-9propyl]dimethylmethoxysilane, 2-[methoxy(polyethyleneoxy)6-9propyl]trimethoxysilane, 2-[methoxy(polyethyleneoxy)9-12propyl]trimethoxysilane, 2-[methoxy(polyethyleneoxy)21-24propyl]trimethoxysilane, N,N-bis(2-hydroxyethyl)-3-aminopropyltriethoxysilane, (heptadecafluoro-1,1,2,2-tetrahydrodecyl) trimethoxysilane, 3-(heptafluoroisopropoxy)propyltrimethoxysilane, nonafluorohexyltrimethoxysilane, [perfluoro(polypropyleneoxy)]methoxypropyltrimethoxysilane, 1,3-bis(trifluoropropyl)-1,1,3,3-tetramethyldisilazane, (tridecafluoro-1,1,2,2-tetrahydrooctyl)dimethylchlorosilane, N,N-bis(2-hydroxyethyl)-3-aminopropyltriethoxysilane, bis(methoxyethyl)-3-trimethoxysilylpropylammoniumchloride, carboxyethylsilanetriol disodium salt, and 3-(trihydroxysilyl)-1-propanesulfonic acid; and
the respective passivation component attached to the polymeric hydrogel is selected from the group consisting of dibenzocyclooctyne-poly(ethylene glycol)-carboxylic acid, norbornene-poly(ethylene glycol)-carboxylic acid, bicyclo[6.1.0]non-4-yne-poly(ethylene glycol)-carboxylic acid, dibenzocyclooctyne-poly(ethylene glycol)-amine, norbornene-poly(ethylene glycol)-amine, bicyclo[6.1.0]non-4-yne-poly(ethylene glycol)-amine, dibenzocyclooctyne-poly(ethylene glycol)-hydroxyl, norbornene-poly(ethylene glycol)-hydroxyl, bicyclo[6.1.0]non-4-yne-poly(ethylene glycol)-hydroxyl, dibenzocyclooctyne-poly(ethylene glycol)-sulfonic acid, norbornene-poly(ethylene glycol)-sulfonic acid, and bicyclo[6.1.0]non-4-yne-poly(ethylene glycol)-sulfonic acid.
13 . The flow cell as defined in claim 1 , wherein:
the respective passivation components are attached to each of the interstitial regions and the polymeric hydrogel;
the third end group and the fourth end group are each a zwitterionic end group selected from the group consisting of phosphocholine, a sulfobetaine, and a carboxybetaine;
the third linker is poly(ethylene glycol) and further includes a silane functional group attached to the interstitial regions; and
the fourth linker is poly(ethylene glycol)-dibenzocyclooctyne, poly(ethylene glycol)-norbornene, bicyclo[6.1.0]non-4-yne-poly(ethylene glycol), bicyclo[6.1.0]non-4-yne-poly(ethylene glycol) methyl ether, or a combination of poly(ethylene glycol) and pentaerythritol.
14 . The flow cell as defined in claim 1 , wherein:
the respective passivation components are attached to each of the interstitial regions and the polymeric hydrogel;
the third end group and the fourth end group are each the amphoteric polymer end group;
the amphoteric polymer end group includes both a carboxylic acid functional group and an amine functional group;
the third linker is poly(ethylene glycol) and further includes a silane functional group attached to the interstitial regions; and
the fourth linker is poly(ethylene glycol)-dibenzocyclooctyne or poly(ethylene glycol)-norbornene, bicyclo[6.1.0]non-4-yne-poly(ethylene glycol), bicyclo[6.1.0]non-4-yne-poly(ethylene glycol) methyl ether, or a combination of poly(ethylene glycol) and pentaerythritol.
15 .- 32 . (canceled)