Method and apparatus for separation of particles in a microfluidic device

• US 7,094,354 B2
• Filed: 12/19/2002
• Issued: 08/22/2006
• Est. Priority Date: 12/19/2002
• Status: Active Grant

• Current Assignee: Siemens Healthcare Diagnostics Incorporated (Siemens AG)
• Original Assignee: Bayer Healthcare LLC (Bayer AG)
• Inventors: Peters, Ralf-Peter, Schulman, Lloyd, Blankenstein, Gert, Pugia, Michael J., Profitt, James A.
• Primary Examiner(s): Reifsnyder, David A.
• Application Number:
  US10/326,156
• Publication Number:
  US 20040121449A1
• Time in Patent Office: 1,342 Days
• Field of Search: 210/787, 210/198.1, 422/64, 422/67, 422/72, 422/103, 435/2, 435/287.1, 436/45
• US Class Current: 210/787
• CPC Class Codes: B01D 21/262 by using a centrifuge
• CPC Class Codes: B01D 2221/10 Separation devices for use in medical, pharmaceutical or laboratory applications, e.g. separating amalgam from dental treatment residues apparatus for dental treatment A61C17/065
• CPC Class Codes: B01L 2200/0668 Trapping microscopic beads
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/161 Control and use of surface tension forces, e.g. hydrophobic, hydrophilic
• CPC Class Codes: B01L 2400/0406 capillary forces
• CPC Class Codes: B01L 2400/0409 centrifugal forces
• CPC Class Codes: B01L 2400/0688 surface tension valves, capillary stop, capillary break
• CPC Class Codes: B01L 2400/088 by specific surface properties
• CPC Class Codes: B01L 3/502723 characterised by venting arrangements
• CPC Class Codes: B01L 3/502753 characterised by bulk separation arrangements on lab-on-a-chip devices, e.g. for filtration or centrifugation separation in general B01D; microapparatus for analysis using electrophoresis G01N27/44791; sample preparation G01N1/28
• CPC Class Codes: G01N 33/491 by separating the blood components G01N15/05 takes precedence
• CPC Class Codes: Y10T 436/111666 Utilizing a centrifuge or compartmented rotor

Abstract

A microfluidic device provides separation of particles in a liquid sample, particularly, separation of a sample of whole blood into its components for further analysis. Separation into red blood cells and plasma occurs within a few seconds after the blood sample has been transferred into a separation chamber with the application of centrifugal force of less than about five times gravity. With the application of greater force measurement of hematocrit is possible.

Claims

1. A microfluidic device for separating a mixture of a liquid and particles subjecting said mixture to low forces comprising:

(a) an entry port for receiving a sample of said mixture of a liquid and particles;

(b) a capillary passageway in fluid communication by capillary forces with said entry port, optionally containing a hydrophobic or hydrophilic stop;

(c) an elongated separation chamber having proximal and distal ends, said proximal end being in fluid communication by capillary forces with said capillary passageway of (b), and said distal end being closed, said separation chamber comprising

(1) a volume for receiving at least a portion of said sample;

(2) a depth to particle diameter ratio of about 5/1 to 100/1, said ratio being chosen to avoid clogging of the chamber with said particles and as a function of the force to be applied to said mixture; and

(3) a wall surface having a surface energy equal to or less than the surface energy of said particles to facilitate separation of said sample by attraction of said particles to said wall surface;

(d) a vent for removing air from the entry port of (a) and the separation chamber of (c) displaced by said sample.

2. A microfluidic device of claim 1 further comprising an outlet capillary in fluid communication with said separation chamber and disposed to receive said liquid separated from said particles.

3. A microfluidic device of claim 1 wherein said surface energy corresponds to a contact angle of 20–

80°

relative to water.

4. A microfluidic device of claim 1 wherein said separation chamber includes a lower section of said distal end having increased depth to accommodate separated particles.

5. A microfluidic device of claim 1 wherein said separation chamber has length:

width;

depth ratios of about 6;

1;

0.5.

6. A method of separating samples of whole blood comprising:

(a) introducing a sample of whole blood into the entry port of the microfluidic device of claim 1;

(b) rotating said microfluidic device at a predetermined speed with said separation chamber positioned radially outward of the center of rotation relative to said entry port to transfer said whole blood sample into the separation chamber and to separate said whole blood sample into red blood cells and plasma.

7. A method of claim 6 wherein said blood separation takes place in about 1 to 10 seconds with a centrifugal force of less than about 5 times gravity.

8. A method of claim 6 wherein said blood separation takes place within about 2 minutes with a centrifugal force sufficient to enable hematocrit to be measured.

9. A method of claim 6 wherein said blood separation takes place in about 15 to 30 seconds with a centrifugal force up to about 16.7 times gravity.

10. A method of separating samples of whole blood comprising:

(a) introducing a sample of whole blood into the entry port of the microfluidic device of claim 1;

(b) transferring said whole blood sample from said entry port to said stop of (b) through said capillary passageway by capillary forces;

(c) moving said sample from said stop through said capillary passageway to said separation chamber of (c) by applying to said sample a force from the group consisting of pressure, vacuum, electroosmosis, and centrifugal force;

(d) separating said blood sample into red blood cells and plasma in said separation chamber by applying a force of less than live times gravity.

11. A method of claim 10 wherein said force of (d) is less than one times gravity.

12. A microfluidic device for separating whole blood into red blood cells and plasma by subjecting said whole blood to low forces comprising:

(a) an entry port for receiving a sample of whole blood;

(b) a hydrophilic capillary passageway in fluid communication by capillary forces with said entry port, said passageway optionally containing a hydrophobic or hydrophilic stop;

(c) an elongated separation chamber having proximal and distal ends, said proximal end being in fluid communication by capillary forces with said capillary passageway of (b), said distal end being closed, said separation chamber comprising;

(1) a volume for receiving at least a portion of said sample,

(2) a depth to red blood cell diameter ratio of about 5/1 to 100/1, said ratio being chosen to avoid clogging of the chamber with said red blood cells and as a function of the force to be applied to said sample; and

(3) a wall surface having a contact angle relative to water of about 20°

to 80°

to facilitate separation of said sample by attraction of said red blood cells to said wall surface;

(d) a vent for removing air from the entry port of (a), and the separation chamber of (c) displaced by said sample.

13. A microfluidic device of claim 12 further comprising an outlet capillary in fluid communication with said separation chamber and disposed to receive plasma separated from said whole blood.

14. A microfluidic device of claim 13 further comprising wells in fluid communication with said outlet capillary for receiving plasma separated from said whole blood.

15. A microfluidic device of claim 14 wherein said wells in communication with said outlet capillary contain reagents for interaction with said plasma.

16. A microfluidic device of claim 15 wherein said reagents precondition said plasma for subsequent reaction to detect presence of an analyte.

17. A microfluidic device of claim 15 wherein said reagents interact with said plasma to detect presence of an analyte.

18. A microfluidic device of claim 13 wherein said outlet capillary has a smaller cross-sectional area than said separation chamber such that said plasma is prevented from entering said capillary until centrifugal force is applied.

19. A microfluidic device of claim 13 wherein a blood sample is separated within 1 to 10 seconds when subjected to a force of less than about 5 times gravity.

20. A microfluidic device of claim 13 wherein a blood sample is separated within about 15 to 30 seconds when subjected to a force of up to about 16.7 times gravity.

21. A microfluidic device of claim 12 wherein said separation chamber has a volume of about 0.2 to 10 μ

L.

22. A microfluidic device of claim 21 wherein said volume is about 0.3 to 2 μ

L.

23. A microfluidic device of claim 12 wherein said separation chamber has a length:

width;

depth ratio of about 6;

1;

0.5.

24. A microfluidic device of claim 23 wherein the separation chamber has a greater depth at the end of said distal section to provide additional volume for separated red blood cells.

25. A microfluidic device for measuring hematocrit of whole blood samples comprising:

(a) an entry port for receiving a sample of whole blood;

(b) a hydrophilic capillary passageway in fluid communication by capillary forces with said entry port, said passageway containing a hydrophobic or hydrophilic stop;

(c) an elongated separation chamber having proximal and distal ends, said proximal end being in fluid communication by capillary forces with said capillary passageway of (b), said distal end being closed, said separation chamber comprising;

(1) a volume for receiving at least a portion of said sample,

(2) depth to red blood cell diameter ratio of about 5/1 to 100/1, said ratio being chosen to avoid clogging of the chamber with red blood cells and as a function of the force to be applied to said sample; and

(3) a wall surface having a contact angle relative to water of about 20°

to 80°

to facilitate separation of said sample by attraction of said red blood cells to said wall surfaces;

(d) a vent from the entry port of (a) and the separation chamber of (c) for removing air displaced by said sample.

26. A microfluidic device of claim 25 wherein said separation chamber has a volume of about 0.2 to 10 gμ

L.

27. A microfluidic device of claim 26 wherein said separation chamber has a volume of about 0.3 to 2 μ

L.

28. A micro fluidic device of claim 25 wherein said separation chamber has length:

width;

depth ratios of about 6;

1;

0.5.

29. A microfluidic device of claim 25 wherein a blood sample is separated within 2 minutes when subjected to a centrifugal force sufficient to enable hematocrit to be measured.