Compact disk based system for separating immunomagnetic bead labeledparticulates and method thereof

• US 20100227379A1
• Filed: 06/25/2009
• Published: 09/09/2010
• Est. Priority Date: 03/03/2009
• Status: Active Grant

• Current Assignee: National Taiwan University (Government of The Republic of China)
• Original Assignee: National Taiwan University (Government of The Republic of China)
• Inventors: Pan, Yu-Cheng, Wo, Andrew M., Chen, Chen-Lin, Chen, Ken-Chao
• Granted Patent: US 8,226,826 B2
• Time in Patent Office: Days
• US Class Current: 435/261
• 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: C12N 1/02 Separating microorganisms from their culture media
• CPC Class Codes: G01N 2035/00495 Centrifuges
• CPC Class Codes: G01N 35/00069 whereby the sample substrate is of the bio-disk type, i.e. having the format of an optical disk
• CPC Class Codes: Y10T 436/111666 Utilizing a centrifuge or compartmented rotor
• CPC Class Codes: Y10T 436/25375 Liberation or purification of sample or separation of material from a sample [e.g., filtering, centrifuging, etc.]

Abstract

Disclosed is a disk based system for separating at least two types of particulates contained in a sample fluid. The system includes a disk-like carrier board and a magnetic attraction unit. The disk-like carrier board forms at least one flow channel structure, which includes an inner reservoir, at least one separation chamber, and at least one outer reservoir arranged in sequence from a geometric center of the disk-like carrier board to an outer circumferential rim of the disk-like carrier board. A method of separation carried out with the system includes introducing the sample fluid into the inner reservoir and then rotating the disk-like carrier board to induce a centrifugal force. The sample fluid contains particulates that are labeled with immunomagnetic beads and the labeled particulates are attracted by the magnetic force generated by the magnetic attraction unit to retain in the inner reservoir or the separation chamber. Particulates not labeled with the immunomagnetic beads and contained in the sample fluid move with the sample fluid to the outer reservoir.

Claims

1. A disk based system for separating at least two types of particulates, which are respectively labeled and not labeled with immunomagnetic beads, contained in a sample fluid, the system comprising:

a disk-like carrier board, which has a geometric center and an outer circumferential rim, the disk-like carrier board forming at least one flow channel structure, each flow channel structure comprising;

an inner reservoir, which is arranged adjacent to the geometric center of the disk-like carrier board for receiving the sample fluid,

at least one separation chamber, which is arranged adjacent to the inner reservoir and is in fluid communication with the inner reservoir through at least one micro flow channel, and

at least one outer reservoir, which is arranged between the outer circumferential rim of the disk-like carrier board and the separation chamber and is in fluid communication with the separation chamber through at least one micro flow channel; and

a magnetic attraction unit, which is arranged atop the disk-like carrier board and adjacent to the inner reservoir and the separation chamber to generate a magnetic force that has a predetermined distribution of intensity and covers the inner reservoir and the separation chamber;

wherein when the disk-like carrier board spins, the sample fluid is caused by a centrifugal force induced by the spinning of the disk-like carrier board to flow from the inner reservoir through the micro flow channels to the outer reservoir whereby the immunomagnetic bead labeled particulates, when passing through the separation chamber, are attracted by the magnetic force generated by the magnetic attraction unit to thereby remain in the separation chamber, and the non-labeled particulates entrain the sample fluid to flow from the inner reservoir through the micro flow channels to the outer reservoir.

2. The system as claimed in claim 1, wherein the disk-like carrier board comprises a cover layer arranged thereon.

3. The system as claimed in claim 1, wherein the disk-like carrier board comprises a base layer and at least one flow channel structure layer, the flow channel structure being formed in the flow channel structure layer.

4. The system as claimed in claim 1, wherein the flow channel structure of the disk is selectively formed by laser machining, CNC machining, micromachining, and injection molding.

5. The system as claimed in claim 1, wherein the outer reservoir has an outer bank that forms at least one collection bin.

6. The system as claimed in claim 1, wherein the outer reservoir comprises at least one pair of reverse flow baffles for preventing reverse flow of the sample fluid.

7. The system as claimed in claim 1, wherein the micro flow channel communicating between the inner reservoir and the separation chamber and the micro flow channel communicating between the separation chamber and the outer reservoir have different channel heights.

8. The system as claimed in claim 1, wherein the micro flow channel communicating between the inner reservoir and the separation chamber and the micro flow channel communicating between the separation chamber and the outer reservoir have different channel widths.

9. The system as claimed in claim 1, wherein the micro flow channel communicating between the inner reservoir and the separation chamber and the micro flow channel communicating between the separation chamber and the outer reservoir have different channel lengths.

10. The system as claimed in claim 1, wherein the flow channel structure of the disk-like carrier board comprises at least one fluid extraction passage in fluid communication with the outer reservoir to allow the sample fluid to be discharged through the fluid extraction passage.

11. The system as claimed in claim 10, wherein the fluid extraction passage comprises a section extending from the outer reservoir in a direction toward the geometric center of the disk-like carrier board.

12. The system as claimed in claim 10, wherein the fluid extraction passage is set in fluid communication with at least one fluid collection chamber for collecting the sample fluid discharged through the fluid extraction passage.

13. The system as claimed in claim 10, wherein the fluid extraction passage comprises a valve mounted thereto.

14. The system as claimed in claim 1, wherein the magnetic attraction unit comprises a magnetic element selected from the group consisting of permanent magnet and electromagnet.

15. The system as claimed in claim 1, wherein the magnetic attraction unit comprises a plurality of magnetic elements to generate a magnetic force that has a high gradient distribution of magnetic field.

16. A method for separating particulates labeled with immunomagnetic beads, wherein a disk-like carrier board forms at least one flow channel structure thereon, each flow channel structure comprising an inner reservoir arranged adjacent to a geometric center of the disk-like carrier board, at least one separation chamber arranged in fluid communication with the inner reservoir through at least one micro flow channel, and at least one outer reservoir arranged between an outer circumferential rim of the disk-like carrier board and the separation chamber and in fluid communication with the separation chamber through at least one micro flow channel, a magnetic attraction unit being arranged atop the disk-like carrier board and adjacent to the inner reservoir and the separation chamber to generate a magnetic force that has a predetermined distribution of intensity and covers the inner reservoir and the separation chamber, the method comprising the following steps:

(a) introducing a sample fluid containing at least two types of particulates, which are respectively labeled and not labeled with immunomagnetic beads, into the inner reservoir;

(b) spinning the disk-like carrier board with a first predetermined rotation speed so that the sample fluid is caused by a centrifugal force induced by the spinning of the disk-like carrier board to flow from the inner reservoir through the micro flow channel into the separation chamber with a portion of the immunomagnetic-bead-labeled particulates is attracted by the magnetic force that is generated by the magnetic attraction unit and covers the inner reservoir to retain in the inner reservoir;

(c) subjecting a remaining portion of the immunomagnetic bead labeled particulates to attraction by the magnetic force that is generated by the magnetic attraction unit and covers the separation chamber to retain in the separation chamber; and

(d) collecting the non-immunomagnetic-bead-labeled particulates, which entrain the sample fluid through the micro flow channel, in the outer reservoir.

17. The method as claimed in claim 16 further comprising, before step (d), a step of spinning the disk-like carrier board with a second predetermined rotational speed to cause the sample fluid to flow from the separation chamber through the micro flow channel into the outer reservoir.

18. The method as claimed in claim 16 further comprising, after step (d), a step of spinning the disk-like carrier board with a third predetermined rotational speed to cause the non-immunomagnetic bead labeled particulates contained in the sample fluid to be collected in at least one collection bin formed in an outer bank of the outer reservoir.

19. The method as claimed in claim 16 further comprising, after step (d), a step of keeping the disk-like carrier board still to have the sample fluid in the outer reservoir filled into at least one fluid extraction passage that communicates with the outer reservoir.

20. The method as claimed in claim 19 further comprising, after the step of filling the sample fluid of the outer reservoir into the at least one fluid extraction passage communicating the outer reservoir, a step of spinning the disk-like carrier board with a fourth predetermined rotational speed to have the sample fluid flowing through the fluid extraction passage into at least one fluid collection chamber in fluid communication with the fluid extraction passage for separation from the non-immunomagnetic bead labeled particulates.

21. The method as claimed in claim 16 further comprising, after step (d),a step of vibrating the disk-like carrier board to have the non-immunomagnetic bead labeled particulates re-arranged.

22. The method as claimed in claim 21, wherein the vibration of the disk-like carrier board is realized through ultrasonic cleaning in order to have the non-immunomagnetic bead labeled particulates re-arranged.