Sonication to selectively lyse different cell types

• US 20060134616A1
• Filed: 02/09/2006
• Published: 06/22/2006
• Est. Priority Date: 09/19/2003
• Status: Active Grant

• Current Assignee: MicroFluidic Systems, Inc.
• Original Assignee: MicroFluidic Systems, Inc.
• Inventors: Yuan, Bob, Belgrader, Phillip I.
• Granted Patent: US 7,785,869 B2
• Time in Patent Office: Days
• US Class Current: 435/5
• CPC Class Codes: B01L 2200/0631 Purification arrangements, e.g. solid phase extraction [SPE]
• CPC Class Codes: B01L 2200/10 Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
• CPC Class Codes: B01L 2300/0636 Integrated biosensor, microarrays
• CPC Class Codes: B01L 2300/0864 comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
• CPC Class Codes: B01L 2300/0867 Multiple inlets and one sample wells, e.g. mixing, dilution
• CPC Class Codes: B01L 2300/087 Multiple sequential chambers
• CPC Class Codes: B01L 2400/0409 centrifugal forces
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 3/5025 for parallel transport of multiple samples
• CPC Class Codes: B01L 3/5027 by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip B01L3/5023 takes precedence; micromixers B01F33/30; microreactors for synthesis B01J19/0093; microcapillary devices in general B81B1/00
• CPC Class Codes: C12M 47/06 Hydrolysis; Cell lysis; Extraction of intracellular or cell wall material lysis of microorganisms C12N1/06; extracting or separating nucleic acids from biological samples C12N15/1003
• CPC Class Codes: C12N 1/06 Lysis of microorganisms

Abstract

A sonication apparatus is directed to a microfluidic-based system to automate differential extraction of specific cell types within a mixed sample. The microfluidic-based system includes a sonication module for selective cell lysis, separating means to eliminate centrifugation, high surface area pillar chip modules to purify DNA from a cell lysate, and microfluidic circuitry to integrate the steps in an automated platform.

Claims

1-38. -38. (canceled)

39. An apparatus to selectively lyse a specific type of cell, the apparatus comprising:

a. a sonication chamber to receive a sample and to provide a first sonication energy, the sample having at least two different cell types, wherein a first cell type is lysed at the first sonication energy to form a first lysate;

b. separating means to separate the first lysate from the sample;

c. a collection vessel to collect the first lysate; and

d. microfluidic circuitry to couple the sonication chamber, the separating means, and the collection vessel.

40. The apparatus of claim 39 wherein the first cell type comprises an epithelial cell.

41. The apparatus of claim 39 wherein the first cell type comprises a virus, a microbe, or a spore.

42. The apparatus of claim 39 wherein the separating means passes the first lysate and blocks the remaining sample.

43. The apparatus of claim 42 wherein the remaining sample includes a second cell type, the second cell type is lysed at a second sonication energy.

44. The apparatus of claim 43 wherein microfluidic circuitry provides the remaining sample with the second cell type to the sonication chamber to be lysed.

45. The apparatus of claim 43 wherein the sonication chamber provides the second sonication energy to lyse the second cell type, thereby forming a second lysate.

46. The apparatus of claim 45 further comprising a purification chip coupled to the sonication chamber to receive the second lysate and collect a DNA from the second lysate.

47. The apparatus of claim 46 wherein the apparatus is automated.

48. The apparatus of claim 43 wherein the second cell type comprises a sperm cell.

49. The apparatus of claim 43 further comprising a second sonication chamber such that the microfluidic circuitry provides the remaining sample with the second cell type to the second sonication chamber to be lysed.

50. The apparatus of claim 49 wherein the second sonication chamber provides the second sonication energy to lyse the second cell type, thereby forming a second lysate.

51. The apparatus of claim 50 further comprising a purification chip coupled to the second sonication chamber to receive the second lysate and collect a DNA from the second lysate.

52. The apparatus of claim 51 wherein the apparatus is automated.

53. The apparatus of claim 39 further comprising a purification chip to receive the lysate and collect a DNA from the lysate.

54. The apparatus of claim 50 wherein the apparatus is automated.

55. The apparatus of claim 39 wherein the sample includes N different cell types, each cell type is lysed at a different sonication energy.

56. The apparatus of claim 55 further comprising N-1 or less additional sonication chambers to apply N-1 or less different sonication energies to the corresponding N-1 or less different cell types.

57. The apparatus of claim 56 further comprising N or less purification chips to receive N or less lysates from the N or less sonication chambers.

58. The apparatus of claim 39 wherein the separating means comprises means for performing centrifugation on the sample to form a sperm cell pellet.

59. The apparatus of claim 39 wherein the sonication chamber provides a third sonication energy to a sample matrix, thereby releasing the sample from the sample matrix.