Sonication to selectively lyse different cell types

• US 7,785,869 B2
• Filed: 02/09/2006
• Issued: 08/31/2010
• Est. Priority Date: 09/19/2003
• Status: Expired due to Fees

• Current Assignee: MicroFluidic Systems, Inc.
• Original Assignee: MicroFluidic Systems, Inc.
• Inventors: Belgrader, Phillip I., Yuan, Bob
• Primary Examiner(s): Beisner; William H
• Application Number:
  US11/352,108
• Publication Number:
  US 20060134616A1
• Time in Patent Office: 1,664 Days
• Field of Search: 435/288.5, 435/306.1, 366/127, 241/2
• US Class Current: 435/306.1
• 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. 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, further wherein the sonication chamber is configured to selectively lyse the first cell type, thereby forming a first lysate, while leaving one or more non-lysed cell types within a remaining sample;

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, wherein the microfluidic circuitry is configured to enable back-flow capability for moving the remaining sample including the one or more non-lysed cell types from the separating means to the sonication chamber, further wherein the sonication chamber is configured to lyse a second cell type within the remaining sample.

2. The apparatus of claim 1 wherein the first cell type comprises an epithelial cell.

3. The apparatus of claim 1 wherein the first cell type comprises a virus, a microbe, or a spore.

4. The apparatus of claim 1 wherein the separating means passes the first lysate and blocks the remaining sample.

5. The apparatus of claim 4 wherein the second cell type is lysed at a second sonication energy.

6. The apparatus of claim 5 wherein the sonication chamber provides the second sonication energy to lyse the second cell type, thereby forming a second lysate.

7. The apparatus of claim 6 further comprising a purification chip coupled to the sonication chamber to receive the second lysate and collect a DNA from the second lysate.

8. The apparatus of claim 7 wherein the apparatus is automated.

9. The apparatus of claim 5 wherein the second cell type comprises a sperm cell.

10. The apparatus of claim 1 further comprising a purification chip to receive the first lysate and collect a DNA from the first lysate.

11. The apparatus of claim 10 wherein the apparatus is automated.

12. The apparatus of claim 1 wherein the sample includes N different cell types, each cell type is lysed at a different sonication energy.

13. The apparatus of claim 12 wherein the sonication chamber is configured to apply N different sonication energies to the corresponding N different cell types.

14. The apparatus of claim 13 further comprising N purification chips to receive N lysates from the sonication chamber.

15. The apparatus of claim 1 wherein the separating means comprises means for performing centrifugation on the sample to form a sperm cell pellet.

16. The apparatus of claim 1 wherein the sonication chamber provides a third sonication energy to a sample matrix, thereby releasing the sample from the sample matrix.