Devices including osmotically balanced barriers, and methods of making and using the same

Patent metadata

Field	Value
Publication number	US-2025223637-A1
Application number	US-202318850831-A
Country	US
Kind code	A1
Filing date	Mar 29, 2023
Priority date	Mar 31, 2022
Publication date	Jul 10, 2025
Grant date	—

Abstract

Devices including osmotically balanced barriers, and methods of making and using the same, are provided herein. A fluidic well may include a barrier having first and second sides. A first fluid within the fluidic well may contact the first side of the barrier, and may have a first composition including a first concentration of a salt. A second fluid within the fluidic well may contact the second side of the barrier and may have a second composition including a second concentration of the salt that is different than the first concentration. The difference between the first and second concentrations of the salt may generate a first osmotic pressure across the barrier. The second composition further may include a concentration of a compound other than the salt. The concentration of the compound may generate a second osmotic pressure across the barrier that opposes and substantially balances the first osmotic pressure.

First claim

1 . A device, comprising:
a fluidic well comprising a barrier, the barrier comprising a first side and a second side; a first fluid within the fluidic well and in contact with the first side of the barrier, the first fluid having a first composition comprising a first concentration of a salt; and a second fluid within the fluidic well and in contact with the second side of the barrier, the second fluid having a second composition comprising a second concentration of the salt that is different than the first concentration of the salt, a difference between the first and second concentrations of the salt generating a first osmotic pressure across the barrier, the second composition further comprising a concentration of a compound other than the salt, the concentration of the compound generating a second osmotic pressure across the barrier that opposes and substantially balances the first osmotic pressure.

2 . The device of claim 1 , wherein the first concentration of the salt is between about 1.1 and about 50 times the second concentration of the salt.

3 - 4 . (canceled)

5 . The device of claim 1 , wherein the first concentration of the salt is above about 150 mM, and wherein the second concentration of the salt is below about 100 mM.

6 . (canceled)

7 . The device of claim 1 , wherein the first composition substantially does not include the compound.

8 . The device of claim 1 , wherein the concentration of the compound is between about 1.1 and about 50 times the first concentration of the salt.

9 - 10 . (canceled)

11 . The device of claim 1 , wherein the concentration of the compound is above about 100 mM.

12 . The device of claim 1 , wherein the compound is charge neutral.

13 . The device of claim 1 , wherein the compound increases viscosity of the second fluid.

14 . The device of claim 1 , wherein the compound comprises an alcohol, a protein, or a polysaccharide.

15 - 16 . (canceled)

17 . The device of claim 14 , wherein the polysaccharide comprises trehalose or a cyclodextrin.

18 - 20 . (canceled)

21 . The device of claim 1 , wherein:
the first composition comprises a first concentration of an aqueous buffer, the second composition comprises a second concentration of the aqueous buffer, and the first concentration of the aqueous buffer is approximately equal to the second concentration of the aqueous buffer.

22 . The device of claim 1 , further comprising a nanopore disposed within the barrier and providing an aperture fluidically coupling the first side to the second side,
wherein a portion of the salt moves from the second side of the barrier to the first side of the barrier through the aperture.

23 . (canceled)

24 . The device of claim 22 , wherein the compound substantially does not move from the second side of the barrier to the first side of the barrier through the aperture.

25 . The device of claim 22 , wherein the compound is larger in at least one dimension than the aperture.

26 - 29 . (canceled)

30 . The device of claim 29 , further comprising:
a polymerase in the second composition or coupled to the nanopore or the barrier; and first and second polynucleotides, wherein the second composition further comprises a plurality of nucleotides, and the polymerase is for sequentially adding nucleotides of the plurality to the first polynucleotide using a sequence of the second polynucleotide.

31 . The device of claim 30 , further comprising:
a first electrode configured to contact the first fluid; a second electrode configured to contact the second fluid; and circuitry in operable communication with the first and second electrodes and configured to detect changes in an electrical characteristic of the aperture responsive to the polymerase sequentially adding nucleotides of the plurality to the first polynucleotide using a sequence of the second polynucleotide.

32 . A sequencing method, comprising:
using the circuitry of the device of claim 31 to detect changes in the electrical characteristic of the aperture responsive to the polymerase sequentially adding nucleotides of the plurality to the first polynucleotide using a sequence of the second polynucleotide.

33 . (canceled)

34 . A device, comprising:
a fluidic well comprising a barrier, the barrier comprising a first side and a second side; a first fluidic reservoir having a first amount of a salt therein; a second fluidic reservoir having a second amount of the salt, and an amount of a compound, therein; at least one fluidic channel for:
receiving a first solvent in the first fluidic reservoir such that the first solvent dissolves the first amount of the salt to form a first composition;
contacting the first side of the barrier with the first composition;
receiving a second solvent in the second fluidic reservoir such that the second solvent dissolves the second amount of the salt and the amount of the compound to form a second composition; and
contacting the second side of the barrier with the second composition,
wherein the second concentration of the salt in the second composition is different than the first concentration of the salt in the first composition so as to generate a first osmotic pressure across the barrier, and wherein the concentration of the compound in the second composition generates a second osmotic pressure across the barrier that opposes and substantially balances the first osmotic pressure.

35 . (canceled)

36 . The device of claim 34 , further comprising:
the first and second solvents; the first and second compositions; and a nanopore disposed within the barrier and providing an aperture fluidically coupling the first side to the second side, wherein a portion of the salt in the second composition moves from the second side of the barrier to the first side of the barrier through the aperture.

37 - 39 . (canceled)

40 . The device of claim 36 , wherein the compound in the second composition substantially does not move from the second side of the barrier to the first side of the barrier through the aperture.

41 - 43 . (canceled)

44 . The device of claim 36 , further comprising:
a polymerase in the second composition or coupled to the nanopore or the barrier; and first and second polynucleotides.

45 . The device of claim 44 , wherein the second composition further comprises a plurality of nucleotides, and wherein the polymerase is for sequentially adding nucleotides of the plurality to the first polynucleotide using a sequence of the second polynucleotide.

46 . The device of claim 45 , further comprising:
a first electrode configured to contact the first composition; a second electrode configured to contact the second composition; and circuitry in operable communication with the first and second electrodes and configured to detect changes in an electrical characteristic of the aperture that are responsive to the polymerase sequentially adding nucleotides of the plurality to the first polynucleotide using a sequence of the second polynucleotide.

47 . A sequencing method, comprising:
using the circuitry of the device of claim 46 to detect changes in the electrical characteristic of the aperture that are responsive to the polymerase sequential

Assignees

• Illumina Inc

Inventors

• Von Hatten Xavier
• Conde-Gonzalez Antonio
• Kocsis Istvan
• Richez Alexandre

Classifications

• C12Q1/48

  involving transferase · CPC title

• C12N9/1241

  Nucleotidyltransferases (2.7.7) · CPC title

• G01N33/48721

  Investigating individual macromolecules, e.g. by translocation through nanopores (Coulter counters in general G01N15/12; fabrication methods for nanoscale apertures B81B1/00; sequencing of nucleic acids C12Q1/68) · CPC title

• C12Q1/6869 Primary

  Methods for sequencing · CPC title