Microfluidic system for detecting a biological entity in a sample

• US 8,372,657 B2
• Filed: 10/20/2010
• Issued: 02/12/2013
• Est. Priority Date: 10/20/2009
• Status: Expired due to Fees

• Current Assignee: Agency For Science Technology and Research
• Original Assignee: Agency For Science Technology and Research
• Inventors: Reboud, Julien, Kao, Linus Tzu-Hsiang, Chen, Yu, Wang, Yan Ping, Liaw, Janice Hao Yuan, Tang, Kum Cheong, Chung, Andre Yao-Kuang, Ng, Shin Yun
• Primary Examiner(s): Wallenhorst, Maureen
• Application Number:
  US12/908,561
• Publication Number:
  US 20110129931A1
• Time in Patent Office: 846 Days
• Field of Search: 436/501, 436/518, 436/524, 436/531, 436/532, 436/63, 436/71, 436/149, 436/150, 436/174, 436/175, 436/177, 436/178, 436/180, 435/2, 435/4, 435/5, 435/6.1, 435/7.1, 435/29, 435/30, 435/34, 435/325, 435/326, 435/366, 435/372, 435/372.2, 435/372.3, 435/287.1, 435/287.2, 435/287.3, 435/288.3, 435/288.4, 435/288.5, 422/82.01, 422/82.02, 422/502, 422/503, 422/504, 422/507, 422/513, 422/527, 422/534, 422/535, 422/68.1, 506/39
• US Class Current: 436/180
• CPC Class Codes: B01L 2200/0668 Trapping microscopic beads
• CPC Class Codes: B01L 2300/0636 Integrated biosensor, microarrays
• CPC Class Codes: B01L 2300/0645 Electrodes
• CPC Class Codes: B01L 2300/0681 Filter
• 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 33/54366 Apparatus specially adapted for solid-phase testing
• CPC Class Codes: Y10T 436/25 including sample preparation
• CPC Class Codes: Y10T 436/25375 Liberation or purification of sample or separation of material from a sample [e.g., filtering, centrifuging, etc.]
• CPC Class Codes: Y10T 436/255 including use of a solid sorbent, semipermeable membrane, or liquid extraction
• CPC Class Codes: Y10T 436/2575 Volumetric liquid transfer

Abstract

According to various embodiments, a microfluidic system for detecting a biological entity in a sample volume is provided. The microfluidic system may include: a chamber configured to receive the sample volume, wherein the chamber includes a detection region for detecting the biological entity; a first port in fluid communication with the chamber; and a second port including a filter in fluid communication with the chamber; and wherein a fluid provided to the first port or the second port flows between the first port and the second port through the chamber.

Claims

1. A microfluidic system for detecting a biological entity in a sample volume, the microfluidic system comprising:

a chamber configured to receive the sample volume, wherein the chamber comprises a detection region for detecting the biological entity;

a first port in fluid communication with the chamber; and

a second port comprising a filter in fluid communication with the chamber; and

wherein a fluid provided to the first port or the second port flows between the first port and the second port through the chamber such that the fluid provided to the first port passes the sample volume through the detection region and the filter to retain the biological entity at the filter, and the fluid provided to the second port transfers the biological entity retained at the filter to the detection region.

2. The microfluidic system according to claim 1, further comprising:

at least one microchannel configured to couple the first port to the chamber; and

at least one microchannel configured to couple the second port to the chamber.

3. The microfluidic system according to claim 1, further comprising a third or more ports in fluid communication with the chamber.

4. The microfluidic system according to claim 1, wherein the detection region is configured to detect the biological entity using a label-free detection method.

5. The microfluidic system according to claim 1, wherein the detection region comprises a microelectrode array.

6. The microfluidic system according to claim 5, further comprising capture molecules configured to attach to a surface of the microelectrode array.

7. The microfluidic system according to claim 6, wherein the fluid provided to the first port is configured to flow through the chamber and the filter such that the biological entity is retained by the filter and the fluid provided to the second port is configured to flow through the filter such that the biological entity is removed from the filter to the chamber for capture by the capture molecules attached to the surface of the microelectrode array.

8. The microfluidic system according to claim 5, wherein the microelectrode array is configured to generate a dielectrophoretic force.

9. The microfluidic system according to claim 5, further comprising a microchip being formed on or in the microelectrode array.

10. The microfluidic system according to claim 1, wherein the chamber comprises an open chamber.

11. The microfluidic system according to claim 1, wherein the filter comprises a membrane.

12. The microfluidic system according to claim 1, further comprising a movably arranged magnetic element arranged below the detection region, the magnetic element being configured to provide a magnetic field in a vicinity of the detection region.

13. The microfluidic system according to claim 1, wherein the biological entity is selected from the group consisting of a biomarker, a cell, a eukaryotic cell, a prokaryotic cell, a mammalian cell, a yeast cell, a tumour cell, a circulating tumor cell, a blood cell, a peripheral blood mononuclear cell, a cell of an immune system, a white blood cell, a T cell, a T helper cell, a lymphocyte, a CD4 lymphocyte, a progenitor cell, an endothelial progenitor cell, a fetal cell, an organelle, a virus particle, a biopolymer, a polypeptide, a nucleic acid, a lipid, an oligosaccharide, and any combination thereof.

14. A method for manufacturing a microfluidic system for detecting a biological entity in a sample volume, the method comprising:

providing a chamber configured to receive the sample volume, wherein the chamber comprises a detection region for detecting the biological entity;

providing a first port in fluid communication with the chamber; and

providing a second port comprising a filter in fluid communication with the chamber; and

wherein a fluid provided to the first port or the second port flows between the first port and the second port through the chamber such that the fluid provided to the first port passes the sample volume through the detection region and the filter to retain the biological entity at the filter, and the fluid provided to the second port transfers the biological entity retained at the filter to the detection region.

15. A method for detecting a biological entity in a sample volume using a microfluidic system for detecting a biological entity in a sample volume, the microfluidic system comprising:

a chamber configured to receive the sample volume, wherein the chamber comprises a detection region for detecting the biological entity;

a first port in fluid communication with the chamber; and

a second port comprising a filter in fluid communication with the chamber; and

wherein a fluid provided to the first port or the second port flows between the first port and the second port through the chamber;

the method comprising;

providing the sample volume to the chamber;

providing the fluid to the first port to pass the sample volume through the detection region and the filter to retain the biological entity at the filter;

providing the fluid to the second port to transfer the biological entity retained at the filter to the detection region of the chamber; and

detecting the biological entity.

16. The method according to claim 15, further comprising trapping the biological entity in a vicinity of the detection region via a movably arranged magnetic element arranged below the detection region.

17. The method according to claim 16, wherein the sample volume comprises:

a blood sample; and

a lysis buffer solution; and

wherein the lysis buffer solution comprises;

a lysing agent;

a pH buffer; and

an anti-coagulant.

18. The method according to claim 17, wherein the lysis buffer solution further comprises a plurality of magnetic beads for coupling to the biological entity.

19. The method according to claim 18, wherein the lysing agent comprises ammonium chloride, the pH buffer comprises sodium bicarbonate and the anti-coagulant comprises ethylenediaminetetraacetic acid.

20. The method according to claim 19, wherein a concentration of the ammonium chloride is about 10 mM to about 150 mM.

21. The method according to claim 20, wherein a volume ratio of the ammonium chloride to the blood sample is between 1:

1 to 1;

10.

22. The method according to claim 21, wherein a concentration of the sodium bicarbonate is between about 10 mM to about 100 mM.

23. The method according to claim 22, wherein a concentration of the ethylenediaminetetraacetic acid is between about 0.01 mM to about 1.0 mM or about 0.1 mM.

24. The method according to claim 15, wherein providing the fluid to the first port to pass the sample volume through the filter to retain the biological entity is repeated at least one time.

25. The method according to claim 15, wherein transferring the biological entity retained at the filter to the detection region of the chamber is repeated at least one time.

26. The method according to claim 15, wherein detecting the biological entity comprises:

incubating the biological entity; and

performing measurements for detecting the biological entity.

27. The method according to claim 15, wherein detecting the biological entity comprises a process selected from the group consisting of dielectrophoresis, capturing by capture molecules, impedance measuring, and any combination thereof.

28. The method according to claim 15, wherein the fluid comprises a buffer solution.