Biochips including ion transport detecting strucutres and methods of use

• US 20020182627A1
• Filed: 03/22/2002
• Published: 12/05/2002
• Est. Priority Date: 03/24/2001
• Status: Active Grant

• Current Assignee: AVIVA Biosciences Corporation
• Original Assignee: AVIVA Biosciences Corporation
• Inventors: Wang, Xiaobo, Wu, Lei, Cheng, Jing, Yang, Weiping, Xu, Jia, Xu, Jun Quan, Huang, Ming Xiang
• Granted Patent: US 7,968,305 B2
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: B01L 2200/06 Fluid handling related problems
• CPC Class Codes: B01L 2300/0627 Sensor or part of a sensor is integrated
• CPC Class Codes: B01L 2300/0645 Electrodes
• CPC Class Codes: B01L 2300/0819 Microarrays; Biochips
• CPC Class Codes: B01L 2400/0415 electrical forces, e.g. electrokinetic
• CPC Class Codes: B01L 2400/043 magnetic forces
• CPC Class Codes: B01L 2400/0436 acoustic forces, e.g. surface acoustic waves [SAW]
• CPC Class Codes: B01L 2400/0457 passive flow or gravitation
• CPC Class Codes: B01L 2400/049 vacuum
• CPC Class Codes: B01L 3/50273 characterised by the means or forces applied to move the fluids micropumps F04B19/006, of the membrane type F04B43/043
• CPC Class Codes: B01L 3/502761 specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules investigating characteristics of particles G01N15/00
• CPC Class Codes: B01L 3/502792 for moving individual droplets on a plate, e.g. by locally altering surface tension
• CPC Class Codes: B03C 5/026 using open-gradient differential dielectric separation, i.e. using electrodes of special shapes for non-uniform field creation, e.g. Fluid Integrated Circuit [FIC]
• CPC Class Codes: B03C 5/028 using travelling electric fields, i.e. travelling wave dielectrophoresis [TWD]
• CPC Class Codes: C12M 35/02 Electrical or electromagnetic means, e.g. for electroporation or for cell fusion
• CPC Class Codes: C12M 47/04 Cell isolation or sorting purging biological preparations of unwanted cells C12N5/0081, determining the presence or kind of microorganism C12Q1/04
• CPC Class Codes: C12Q 1/6869 Methods for sequencing
• CPC Class Codes: C12Q 2565/629 being a microfluidic device
• CPC Class Codes: G01N 15/1031 by measuring electrical or magnetic effects
• CPC Class Codes: G01N 33/48728 Investigating individual cells, e.g. by patch clamp, voltage clamp investigating individual particles in general G01N15/10
• CPC Class Codes: G01N 33/5005 : involving human or animal cells immunoassay G01N33/56966; immunoassays of protozoa G01N33/56905; protozoa in screening assays C12Q1/025

Abstract

The present invention recognizes that the determination of ion transport function or property using direct detection methods, such as patch-clamps, whole cell recording or single channel recording, are preferable to methods that utilize indirect detection methods, such as FRET based detection system. The present invention provides biochips and methods of use that allow for the direct analysis of ion transport function or property using microfabricated structures that can allow for automated detection of ion transport function or property. These biochips and methods of use thereof are particularly appropriate for automating the detection of ion transport function or property, particularly for screening purposes.

Claims

1. A biochip comprising:

a) at least one particle positioning means;

b) at least one ion transport measuring means;

wherein said particle positioning means comprises at least one structure selected from the group consisting of dielectric focusing structure, quadropole electrode structure, electrorotation structure, traveling wave dielectrophoresis structure, concentric circular electrode structure, spiral electrode structure, square spiral electrode structure, particle switch structure, dielectrophoresis guide electrode structure, electromagnetic structure and acoustic structure.

2. The biochip of claim 1, wherein said at least one structure is connected to an AC or DC signal source for producing forces on particles introduced onto said biochip to position the said particles to or near the said ion transport measuring means.

4. The biochip of claim 1 or 3, wherein said particle positioning means is active upon a cell or a population of cells.

5. The biochip of claim 1 or 3, wherein said particle positioning means can localize a particle or population of particles at or near said ion transport measuring means.

6. The biochip of claim 1 or 3, wherein said particle positioning means comprises at least two structures selected from the group consisting of dielectric focusing structure, quadropole electrode structure, electrorotation structure, traveling wave dielectrophoresis structure, concentric circular electrode structure, spiral electrode structure, square spiral electrode structure, particle switch structure, electromagnetic structure, acoustic structure and a pressure generation structure,

wherein, when said pressure generation structure is present, said pressure generation structure comprises at least one fluidic channel comprising at least one surface and linked to said ion transport measuring means,

further wherein, when said at least one fluidic channel is present, said at least one surface is electrically charged when said biochip is in contact with a measurement solution and the surface charge distribution of said at least one fluidic channel is capable of, when an appropriate DC electrical field is established in said at least one fluidic channel, producing a pressure near the ion transport measuring means sufficient to move a particle to or near said particle positioning means.

7. The biochip of claim 1 or 3, wherein said particle positioning means comprises a horizontal positioning means.

8. The biochip of claim 1 or 3, wherein said particle positioning means comprises a vertical positioning means.

9. The biochip of claim 1 or 3, wherein said ion transport measuring means comprises a structure suitable for whole cell recording, single channel recording or whole cell recording and single channel recording,

wherein said single channel recording is realized in at least one configuration selected from the group consisting of cell-attached patch configuration and inside-out configuration.

10. The biochip of claim 1 or 3, wherein said ion transport measuring means comprises a hole that extends through said biochip.

11. The biochip of claim 10, further comprising recording electrodes fabricated on the biochip and located on the two surfaces the said hole extends through.

12. The biochip of claim 1 or 3, wherein said ion transport measuring means comprises a capillary.

13. The biochip of claim 1 or 3, wherein said ion transport measuring means comprises at least one recording electrode.

14. The biochip of claim 1 or 3, wherein said ion transport measuring means comprises at least one needle electrode.

15. The biochip of claim 1 or 3, further comprising an additional particle manipulation means.

16. The biochip of claim 1 or 3, further comprising at least one fluidic channel.

17. The biochip of claim 16, wherein said fluidic channels when in combination with external fluidic devices can be used for delivering intracellular solutions, extracellular solutions, cell suspensions, compound solutions to the ion transport measuring means.

18. A device for detecting ion transport activity of a particle, comprising,

at least two fluidic compartments separated by a biochip of claim 1 or claim 3 and operably connected by said ion transport measuring means on said biochip,

at least one fluidic inlet and at least one outlet for one or more of said at least two fluidic compartments.

19. A device of claim 18, wherein said device is made of materials such as glass or plastics.

20. A device of claim 18, further comprising.

a plurality of ion transport measuring means;

a plurality of fluidic compartments located on two sides of the biochip, wherein

each of said ion transport measuring means is connected to two fluidic compartments.

21. A device of claim 18, further comprising fluidic channels for delivering particle suspensions, compound solutions or both to said at least two fluidic compartments.

22. A device of claim 18, further comprising means for connecting an electrical signal source to said particle positioning means on said biochips.

23. A device of claim 18, further comprising means for connecting a measuring device to said at least two fluidic compartments.

24. An apparatus for ion transport measurement, comprising

a device of claim 18,

at least one measuring device in connection with recording electrodes that are in contact with said at least two fluidic chambers in said device,

at least one fluidic device in fluid communication with said inlet and outlet ports on said device,

at least one electrical signal sources connected to said particle positioning means on said biochip.

25. An apparatus of claim 24, wherein said recording electrodes are integral to said biochip and said device further comprising electrical connection pads to operably connect to said recording circuits.

26. An apparatus of claim 24, wherein said recording electrodes are outside said device and are operably in contact with fluidic compartments via inlet and outlet ports of said device.

27. An apparatus of claim 24, wherein said recording circuits comprise a patch clamp amplifier, a data acquisition circuit or both.

28. An apparatus of claim 24, wherein said at least one fluidic device comprises at least one valve for directing or modulating fluid samples to different fluidic paths or rates of flow.

29. An apparatus of claim 24, whereon said at least one fluidic device comprises at least one pump for pumping fluid into and/or out of said fluidic compartments.

30. An apparatus of claim 24, wherein said at least one fluidic device comprises at least one pressure source.

31. A method of detecting ion transport activity of a particle, comprising:

a) contacting at least one sample comprising at least one particle with the biochip of claim 1 or 3;

b) positioning said at least one particle at or near said ion transport transport measuring means; and

c) measuring ion transport activity of said sample or particle using said ion transport transport measuring means.

32. The method of claim 31, wherein said biochip is located in a device of claim 18.

33. The method of claim 31, wherein the said biochip is located in an apparatus of claim 24.

34. The method of claim 31, wherein said at least one sample comprises a biological sample.

35. The method of claim 31, wherein said at least one sample comprises a cell or a population of cells.

36. The method of claim 31, wherein said particle is a biological cell, or a vesicle containing ion transports on the membrane of said vesicle, or a membrane organelle, or a cell fragment.

37. The method of claim 31, wherein said positioning comprising the steps of:

a) applying at least one electrical signal to said a particle positioning means; and

b) monitoring the presence of a particle on the ion transport measuring means.

38. The method of claim 37, wherein said at least one electrical signal is a AC electrical voltage, a DC electrical current or voltage or both.

39. The method of claim 31, wherein said ion transport measuring means is performed in a whole cell configuration, comprising a step of accessing interior of said at least one particle by applying negative pressure pulse, electrical voltage pulses or negative pressure pulse and electrical voltage pulse across the ion transport measuring means, or applying chemical pore formation agents to one fluidic compartment.

40. The method of claim 31, wherein said measuring is performed by cell-attached method, further comprising applying different voltage protocols to the patch membrane and recording electric current responses of the ion transports in the particle.

3. A biochip comprising:

a) at least one ion transport measuring means;

b) at least one particle positioning means,

wherein said particle positioning means comprises at least one fluidic channel linked to said ion transport measuring means,

further wherein a surface of said at least one fluidic channel is electrically charged when said biochip is in contact with a measurement solution and the surface charge distribution of said surface is capable of, when an appropriate electrical field is established in said at least one fluidic channel, producing a pressure near the ion transport measuring means sufficient to move a particle to or near the particle positioning means.

41. A biochip, comprising:

an array of capillaries, wherein members of said array comprise a capillary capable of engaging a particle, cell, vesicle or membrane organelle with a high resistance electrical seal.

42. The biochip of claim 41, wherein members of said array further comprise recording electrodes.

43. A device for detecting ion transport activity of a particle, comprising:

an array of fluidic compartments separated by a biochip of claim 41 and connected by the array of capillaries on the biochip of claim 41, wherein each fluidic compartment comprising at least one fluidic inlet and one outlet for each of said fluidic compartments.

44. An apparatus for ion transport measurement, comprising

at least one device of claim 43,

at least one recording circuit in connection with recording electrodes that are in contact with said at least two fluidic chambers in the device,

at least one fluidic device in fluid communication with said inlet ports and outlet ports on said device.

45. A method of detecting ion transport activity of a particle, comprising:

a) contacting a sample comprising at least one particle with the biochip of claim 41;

b) positioning said at least one particle at or near said capillary; and

c) measuring ion transport activity of said sample or particle using said capillary.

48. An apparatus for ion transport measurement, comprising

a device of claim 47,

recording circuits in connection with said needle electrode structures and recording electrodes that are in contact with said fluidic compartments in the device,

fluidic devices in fluid communication with said inlet and outlet ports on said device.

46. A biochip, comprising:

an array of needle electrodes, wherein said needle electrodes can penetrate a particle and form high resistance electrical seal, wherein said particle is selected from the group consisting of a cell, a vesicle and a membrane organelle.

49. A method of detecting ion transport activity of a particle, comprising:

a) contacting a sample comprising at least one particle with the biochip of claim 46;

b) positioning said at least one particle at said needle electrode; and

c) measuring ion transport activity of said sample or particle using said needle electrodes.

47. A device for detecting ion transport activity of a particle, comprising, an array of fluidic compartments, wherein each fluidic compartment comprising one said needle electrode, at least one fluidic inlet and at least one outlet for each fluidic compartment of said array of fluidic compartments.

50. A biochip, comprising:

an array of ion transport recording units

wherein membrane of said array comprises

a hole through said biochip,

a particle positioning means comprising at least one structure selected

from the group consisting of dielectric focusing structure, quadropole electrode structure, electrorotation structure, traveling wave dielectrophoresis structure, concentric circular electrode structure, spiral electrode structure, square spiral electrode structure, particle switch structure, electromagnetic structure, and acoustic structure.

52. The biochip of claim 50 or claim 51, wherein said particle is a cell or a vesicle.

53. A device for detecting ion transport activity of a particle, comprising:

a biochip of claim 50 or 51 comprising an array of holes,

an array of fluidic compartments on each side of said biochip,

each of said holes connecting two fluidic compartments, and

each fluidic compartment comprising at least one fluidic inlet and at least one outlet for each of said fluidic compartments.

54. An apparatus for ion transport measurement, comprising:

a device of claim 53,

recording circuits in connection with recording electrodes that are in contact with said fluidic compartments in said device,

fluidic devices in fluid communication with said at least one inlet port and said at least one outlet port on said device.

55. A method of detecting ion transport activity of a particle, comprising:

a) contacting a sample comprising at least one particle with the biochip of claim 50 or 51;

b) positioning said at least one particle at or near said ion transport measuring means; and

c) measuring ion transport activity of said sample or particle using said hole.

57. A device for detecting ion transport activity of a particle, comprising a fluidic chip of claim 56, at least one fluidic inlet and one at least one outlet for each of said at least two fluidic channels.

58. An apparatus for ion transport measurement, comprising:

a device of claim 57,

recording circuits in connection with recording electrodes that are in contact with said the fluidic channels in said device,

fluidic devices in fluid communication with said at least one inlet port and at least one outlet port on said device.

51. A biochip, comprising:

an array of ion transport recording units

wherein membrane of said array comprises

a hole through said biochip,

a particle positioning means, comprising at least one fluidic channel operably linked to said ion transport measuring means, wherein the surface of said fluidic channels is electrically charged when the biochip is in contact with measurement solution and the surface charge distribution of said fluidic channels is capable of, when an appropriate electrical field is established in said fluidic channel, producing a pressure inside and/or near the ion transport measuring means sufficient to move a particle away to or near the particle positioning means.

56. A fluidic chip, comprising a substrate:

a) at least two fluidic channels on one side of a substrate;

b) a hole whose smallest diameter is less than 10 micron on side walls of the fluidic channels to connect said at least two fluidic channels,

c) surface of said hole allows engagement with high resistance electric seal a particle selected from the group consisting of cells, biological cells, vesicles, and membrane organelles

59. A method of detecting ion transport activity of a particle, comprising:

a) contacting a sample comprising at least one particle with the fluidic chip of claim 56;

b) engaging said at least one particle at said hole; and

c) measuring ion transport activity of said sample or particle using said hole.