Method and apparatus for separating particles, cells, molecules and particulates

• US 8,292,083 B2
• Filed: 04/18/2008
• Issued: 10/23/2012
• Est. Priority Date: 04/19/2007
• Status: Expired due to Fees

• Current Assignee: Charles Stark Draper Laboratory Incorporated, Children's Medical Center Corporation
• Original Assignee: Charles Stark Draper Laboratory Incorporated, Children's Medical Center Corporation
• Inventors: Varghese, Mathew, Fiering, Jason O., Ingber, Donald E., Xia, Nan, Mescher, Mark J., Borenstein, Jeffrey T., Yung, Chong Wing
• Primary Examiner(s): MATTHEWS, TERRELL HOWARD
• Application Number:
  US12/105,805
• Publication Number:
  US 20090078614A1
• Time in Patent Office: 1,649 Days
• Field of Search: 209/39, 204/450, 204/451, 204/452, 204/454, 204/456
• US Class Current: 209/39
• CPC Class Codes: B03C 1/0332 using permanent magnets
• CPC Class Codes: B03C 2201/18 Magnetic separation whereby the particles are suspended in a liquid
• CPC Class Codes: B03C 2201/26 for use in medical applications
• CPC Class Codes: Y10T 436/2575 Volumetric liquid transfer

Abstract

A method and apparatus for continuously separating or concentrating particles that includes flowing two fluids in laminar flow through a magnetic field gradient which causes target particles to migrate to a waste fluid stream, and collecting each fluid stream after being flowed through the magnetic field gradient.

Claims

1. An apparatus comprising:

a microfluidic flow cell having an upstream end and a downstream end, the flow cell including;

a separation channel;

a first inlet at the upstream end to introduce a first fluid containing particles into the separation channel;

a second inlet at the upstream end to introduce a second fluid into the separation channel in laminar flow with the first fluid;

a first outlet at the downstream end for receiving the first fluid;

a second outlet at the downstream end for receiving the second fluid,

wherein the first inlet and first outlet are formed in a first plane, and the second inlet and the second outlet are formed in a second plane parallel to the first plane;

a magnetic housing including;

a stage for positioning the microfluidic flow cell;

a plate positioned opposite the stage for applying a magnetic field gradient across the separation channel; and

a magnetic source for creating the magnetic field gradient across the plate and the stage.

2. The apparatus of claim 1, wherein the first fluid and the second fluid are positioned relative to each other in the separation channel in the predominant direction of the magnetic field gradient.

3. The apparatus of claim 1, wherein the first fluid and second fluid remain separated until the first fluid and second fluid flow in the same direction.

4. The apparatus of claim 3, comprising a barrier to maintain the separation between the first fluid and the second fluid until the first fluid and the second fluid flow in the same direction.

5. The apparatus of claim 1, wherein the flow cell comprises a plurality of separation channels.

6. The apparatus of claim 5, wherein the separation channels are arrayed laterally with respect to one another across the flow cell, perpendicular to the direction of flow and perpendicular to the predominant direction of the magnetic field gradient.

7. The apparatus of claim 5, wherein the flow cell comprises a plurality of input ports and output ports.

8. The apparatus of claim 5, wherein the flow cell comprises:

a first input port at the upstream end to introduce the first fluid into a respective-first inlet of each of the separation channels;

a second input port at the upstream end to introduce the second fluid into a respective-second inlet of each of the separation channels;

a first output port at the downstream end for receiving the first fluid from the respective-first outlets of each of the separation channels; and

a second output port at the downstream end for receiving the second fluid from the respective-second outlets of each of the separation channels.

9. The apparatus of claim 1, wherein walls of the separation channel of the flow cell include a bio-compatible coating.

10. The apparatus of claim 1, wherein the stage and plate are made of high magnetic permeability metal.

11. The apparatus of claim 1, wherein the surface of the first plate has a shape configured to concentrate the magnetic field gradient at or about the separation channels.

12. The apparatus of claim 1, wherein the separation channel has a cross-section that is circular, oval, or polygonal without sharp corners.

13. The apparatus of claim 1, wherein junctions between the first and second inlets and the separation channel have smooth, rounded transitions to avoid sharp corners, features, or sudden expansions or contractions at the junctions.

14. The apparatus of claim 1, wherein the stage is configured to position a plurality of flow cells stacked with respect to one another in the predominant direction of the magnetic field gradient.

15. The apparatus of claim 14, comprising a plurality of flow cells positioned on the stage.

16. The apparatus of claim 14 comprising a plurality of plates which separate each of the plurality of flow cells from adjacent flow cells.

17. The apparatus of claim 14, wherein each of the plurality of flow cells comprises a plurality of separation channels, and the surface each of the plurality of plates has a shape configured to concentrate the magnetic field gradient at or about each of the plurality of separation channels of each of the plurality of flow cells.

24. The apparatus of claim 1, wherein the magnetic source is generally C-shaped and comprises a first portion and a second portion, wherein the end of the first portion is coupled to the stage and the end of the second portion is coupled to the plate.

25. The apparatus of claim 24, wherein the magnetic housing includes a shim positioned between the uncoupled ends of the first and second portions of the magnetic source, wherein the thickness of the shim can be adjusted to adjust the strength of the magnetic field gradient across the stage and the plate.

26. The apparatus of claim 11, wherein the shape of the surface of the plate includes rectangular, rounded, or prismatic protrusions spaced to align with each of the plurality of separation channels.

27. The apparatus of claim 16, wherein the plurality of plates are made of a magnetically permeable material that concentrate the magnetic field gradient across the plurality of flow cells.

18. An apparatus comprising:

a microfluidic flow cell having an upstream end and a downstream end, the flow cell including;

a separation channel;

a first inlet at the upstream end to introduce a first fluid containing particles into the separation channel;

a second inlet at the upstream end to introduce a second fluid into the separation channel in laminar flow with the first fluid;

a first outlet at the downstream end for receiving the first fluid from the separation channel; and

a second outlet at the downstream end for receiving the second fluid from the separation channel;

wherein the first inlet and first outlet are formed in a first plane, and the second inlet and the second outlet are formed in a second plane parallel to the first plane; and

a magnetic housing including;

a stage for positioning the microfluidic flow cell;

a magnetic element positioned proximate to the separation channel the stage for applying a magnetic field gradient across the separation channel.

19. A method for separating particles from a fluid comprising:

inserting a flow cell into a magnetic housing;

flowing a first fluid containing particles into a separation channel included in of the flow cell;

flowing the second fluid into the separation channel in laminar flow with the first fluid;

applying a magnetic field gradient across the separation channel perpendicular to the direction of flow of the first fluid and the second fluid, whereby at least a portion of particles in the first fluid are caused to migrate into the second fluid;

flowing a portion of the first fluid from the separation channel through a first outlet placed to receive the first fluid;

flowing a portion of the second fluid from the separation channel through a second outlet placed to receive the second fluid, wherein the first inlet and second inlet are formed substantially in a first plane, and the second inlet and the second outlet are formed substantially in a second plane parallel to the first plane; and

removing the flow cell from the magnetic housing.

20. The method of claim 19, wherein the flow cell comprises a plurality of separation channels, and wherein the plurality of separation channels are arrayed laterally with respect to one another across the flow cell, perpendicular to the direction of flow and perpendicular to the predominant direction of the magnetic field gradient.

21. The method of claim 20, comprising coupling paramagnetic particles to the particles in the first fluid prior to flowing the first fluid into at least one of the plurality of separation channels.

22. The method of claim 20, comprising flowing the first fluid into at least one of the separation channels at a different rate from the second fluid.

23. The method of claim 19, wherein the first fluid is blood.

28. The method of claim 19, wherein the magnetic housing includes:

a stage for positioning the flow cell;

a plate positioned opposite the stage for applying the magnetic field gradient across the separation channel; and

a magnetic source for creating the magnetic field gradient across the plate and the stage.

29. The method of claim 28, wherein the magnetic source is generally C-shaped and comprises a first portion and a second portion, wherein the end of the first portion is coupled to the stage and the end of the second portion is coupled to the plate.

30. The method of claim 29, wherein the magnetic housing includes a shim positioned between the uncoupled ends of the first and second portions of the magnetic source, wherein the thickness of the shim can be adjusted to adjust the strength of the magnetic field gradient across the stage and the plate.