Flow cells

Patent metadata

Field	Value
Publication number	US-2022364977-A1
Application number	US-202217727624-A
Country	US
Kind code	A1
Filing date	Apr 22, 2022
Priority date	Apr 26, 2021
Publication date	Nov 17, 2022
Grant date	—

Abstract

An example method includes introducing a first fluid to a flow channel of a flow cell including a working electrode having a surface that is at least partially exposed to the flow channel, the surface being unmodified or modified with a first member of a transition metal complex binding pair, whereby a linking moiety of a complex present in the first fluid chemically attaches the complex to the surface to form a temporarily modified surface of the working electrode; performing a sensing operation involving the complex of the temporarily modified surface; and applying a desorption voltage of the linking moiety to the working electrode, thereby detaching the linking moiety and regenerating the surface.

First claim

1 . A method, comprising:
introducing a first fluid to a flow channel of a flow cell including a working electrode having a surface that is at least partially exposed to the flow channel, the surface being unmodified or modified with a first member of a transition metal complex binding pair, whereby a linking moiety of a complex present in the first fluid chemically attaches the complex to the surface to form a temporarily modified surface of the working electrode; performing a sensing operation involving the complex of the temporarily modified surface; and applying a desorption voltage of the linking moiety to the working electrode, thereby detaching the linking moiety and regenerating the surface.

2 . The method as defined in claim 1 , wherein:
the surface is unmodified; the complex includes the linking moiety and an orthogonal functional group that does not attach to the unmodified surface; and prior to performing the sensing operation, the method further comprises introducing, into the flow channel, a second fluid including a hydrogel having i) a plurality of primers attached thereto and ii) a reactive functional group attached thereto that is reactive with the orthogonal functional group.

3 . The method as defined in claim 2 , wherein the sensing operation involves:
amplifying a template nucleic acid strand using the plurality of primers to generate a cluster of template nucleic acid strands; introducing, into the flow channel, a third fluid including a plurality of optically labeled nucleotides; and optically detecting an incorporation event of a respective one of the plurality of optically labeled nucleotides into a nascent strand along at least some of the template nucleic acid strands.

4 . The method as defined in claim 1 , wherein:
the surface is unmodified; the complex includes the linking moiety and an orthogonal functional group that does not attach to the unmodified surface; and prior to performing the sensing operation, the method further comprises introducing, into the flow channel, a second fluid including a particle having i) a cluster of template nucleic acid strands attached thereto and ii) a reactive functional group attached thereto that is reactive with the orthogonal functional group.

5 . The method as defined in claim 1 , wherein:
the surface is unmodified; the complex includes the linking moiety and a capture oligonucleotide attached to the linking moiety; and prior to performing the sensing operation, the method further comprises introducing, to the flow channel, a second fluid including a particle having i) a cluster of template nucleic acid strands attached thereto and ii) an oligonucleotide attached thereto that is complementary to the capture oligonucleotide.

6 . The method as defined in claim 4 , wherein the sensing operation involves sequencing the cluster of template nucleic acid strands by:
introducing, into the flow channel, a third fluid including a plurality of optically labeled nucleotides; and optically detecting an incorporation event of a respective one of the plurality of optically labeled nucleotides into a nascent strand along at least some of the template nucleic acid strands.

7 . The method as defined in claim 1 , wherein:
the surface is unmodified; and the complex includes a hydrogel having i) the linking moiety attached thereto and ii) a plurality of primers attached thereto.

8 . The method as defined in claim 1 , wherein:
the surface is unmodified; and the complex includes a particle having i) the linking moiety attached thereto and ii) a cluster of template nucleic acid strands attached thereto.

9 . The method as defined in claim 8 , wherein the sensing operation involves sequencing the cluster of template nucleic acid strands by:
introducing, into the flow channel, a second fluid including a plurality of optically labeled nucleotides; and optically detecting an incorporation event of a respective one of the plurality of optically labeled nucleotides into a nascent strand along at least some of the template nucleic acid strands.

10 . The method as defined in claim 1 , wherein:
the surface is modified with the first member of the transition metal complex binding pair; the first member of the transition metal complex binding pair is a transition metal complex; the linking moiety is a ligand, and the ligand is a second member of the transition metal complex binding pair; and the complex includes a metal nanoparticle functionalized with i) the ligand and ii) a hydrogel having a cluster of template nucleic acid strands attached thereto.

11 . The method as defined in claim 1 , wherein:
the surface is modified with the first member of the transition metal complex binding pair; the first member of the transition metal complex binding pair is a ligand; the linking moiety is a transition metal complex, and the transition metal complex is a second member of the transition metal complex binding pair; and the complex includes a hydrogel having i) the transition metal complex attached thereto and ii) a plurality of primers attached thereto.

12 . The method as defined in claim 1 , wherein applying the desorption voltage involves applying a negative bias to the working electrode.

13 . The method as defined in claim 1 , wherein applying the desorption voltage involves applying a positive bias to the working electrode.

14 . The method as defined in claim 1 , further comprising introducing a wash fluid to the flow channel after the desorption voltage is applied.

15 . The method as defined in claim 1 , wherein:
the flow cell includes:
a substrate;
the working electrode positioned over the substrate;
a patterned insulating material positioned over the working electrode, the patterned insulating material defining depressions separated by interstitial regions, wherein the unmodified surface is exposed at each of the depressions; and
a second working electrode positioned over the interstitial regions; and
the method further comprises applying the desorption voltage of the linking moiety to the second working electrode as the first fluid is introduced, thereby repelling the complex from the interstitial regions.

16 . The method as defined in claim 15 , further comprising applying the desorption voltage of the linking moiety to the second working electrode after the sensing operation.

17 . A method, comprising:
introducing a first fluid to a flow channel of a flow cell including a surface of a substrate that is at least partially exposed to the flow channel, the surface being modified with a visible light responsive first member of a transition metal complex binding pair, whereby a linking moiety of a complex present in the first fluid chemically attaches the complex to the surface to form a temporarily modified surface of the substrate; performing a sensing operation involving the complex of the temporarily modified surface; and exposing the temporarily modified surface to visible light, thereby detaching the linking moiety and regenerating the surface.

18 . The method as defined in claim 17 , wherein:
the linking moiety is a thioether; and the complex is a hydrogel having i) the thioether attached thereto and ii) a plurality of primers attached thereto.

19 . The method as defined in claim 17 , wherein:
the linking moiety is a thioether; and the complex is a hydrogel having i) the thioether attached thereto and ii) a cluster of template nucleic acid strands attached thereto.

20 .- 58 . (cancele

Assignees

• Illumina Inc
• Illumina Cambridge Ltd

Inventors

• Black Hayden
• Von Hatten Xavier

Classifications

• G01N15/1404 Primary

  Handling flow, e.g. hydrodynamic focusing · CPC title

• G01N33/5438

  Electrodes · CPC title

• C12Q1/6869 Primary

  Methods for sequencing · CPC title

• C12Q1/6844 Primary

  Nucleic acid amplification reactions · CPC title

• C12Q1/6874

  involving nucleic acid arrays, e.g. sequencing by hybridisation · CPC title