Focusing chamber

• US 8,361,716 B2
• Filed: 10/05/2009
• Issued: 01/29/2013
• Est. Priority Date: 10/03/2008
• Status: Expired due to Fees

• Current Assignee: PathoGenetix, Inc. (Toxic Reports LLC)
• Original Assignee: PathoGenetix, Inc. (Toxic Reports LLC)
• Inventors: Patil, Vishal
• Primary Examiner(s): CROW, ROBERT THOMAS
• Application Number:
  US12/573,758
• Publication Number:
  US 20100112576A1
• Time in Patent Office: 1,212 Days
• Field of Search: 435/6, 435/6.1, 435/7.1, 435/174, 435/283.1, 435/287.1, 435/287.2
• US Class Current: 435/6.1
• CPC Class Codes: C12Q 1/6806 Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay C12Q1/6804 takes precedence
• CPC Class Codes: G01N 1/40 Concentrating samples
• CPC Class Codes: G01N 2001/4088 filtration

Abstract

Aspects of the invention relate to devices and methods of use thereof for concentrating, positioning and/or manipulating agents within a fluid, including but not limited to genomic DNA.

Claims

1. A method of positioning agents in a fluid flow, the method comprising:

providing a chamber having a porous substrate;

flowing a first fluid flow through a first fluid port and through the porous substrate in the chamber, wherein the first fluid flow contains agents;

flowing a second fluid flow through the chamber, the second fluid flow at least partially surrounding the first fluid flow to direct the first fluid flow and any agents contained therein toward a central portion of the porous substrate;

positioning the agents on the central portion of the porous substrate; and

reversing flow through the first fluid port to move any agents positioned on the porous substrate out of the chamber in central streamlines that exit the chamber through the first fluid port.

2. The method of claim 1, wherein flowing the second fluid flow comprises flowing the second fluid flow through one or more second fluid ports positioned about the porous substrate in the chamber, the second fluid flow completely surrounding the first fluid flow to direct the first fluid flow and any agents contained therein toward the central portion of the porous substrate.

3. The method of claim 1, wherein providing the chamber comprises providing a chamber that includes an open top.

4. The method of claim 1, wherein a flow rate of the second fluid flow is five times or greater than a flow rate of the first fluid flow.

5. The method of claim 1, wherein a flow rate of the second fluid flow is fifteen times or greater than a flow rate of the first fluid flow.

6. The method of claim 1, wherein a flow rate of the second fluid flow is twenty times or greater than a flow rate of the first fluid flow.

7. The method of claim 1, wherein agents removed from the chamber are concentrated ten times greater than prior to entering the chamber.

8. The method of claim 1, wherein agents removed from the chamber are concentrated one hundred times greater than prior to entering the chamber.

9. The method of claim 1, wherein the agents are nucleic acids.

10. The method of claim 9, wherein the agents are DNA.

11. The method of claim 9, wherein the nucleic acids maintain 75% to 100% of original length after exiting the chamber.

12. The method of claim 9, wherein the nucleic acids maintain 90% to 100% of original length after exiting the chamber.

13. The method of claim 9, wherein the nucleic acids include DNA having lengths of about 1 megabase in length after exiting the chamber.

14. The method of claim 1, further comprising:

conveying flow with agents positioned in central streamlines from the chamber prior to removing flow from about the central streamlines.

15. The method of claim 14, wherein conveying comprises conveying a fluid a distance greater than 10 cm.

16. The method of claim 14, wherein conveying comprises conveying a fluid a distance greater than 20 cm.

17. The method of claim 16, wherein agents are conveyed without dispersing more than 2 cm in a direction of flow.

18. The method of claim 17, wherein agents are conveyed without dispersing more than 2 cm in a direction of flow.

19. The method of claim 1, wherein flowing the second fluid flow comprises flowing the second fluid flow through one or more second fluid ports positioned about the porous substrate in the chamber.

20. The method of claim 1, further comprising:

removing flow from about the central streamlines to separate fluid flow that lacks agents from the central streamlines that exit the chamber.

21. The method of claim 1, wherein flowing the second fluid flow comprises the second fluid flow completely surrounding the first fluid flow to direct the first fluid flow and any agents contained therein toward the central portion of the porous substrate.

22. The method of claim 3, wherein flowing the first fluid flow comprises flowing the first fluid flow containing agents through a tube that is positioned at the open top on the chamber.

23. The method of claim 1, wherein reversing flow through the first fluid port includes flowing a fluid through a port positioned on a side of the porous substrate opposite the first fluid port such that the fluid flows toward the first fluid port.

24. The method of claim 20, wherein removing flow from about the central streamlines to separate fluid flow that lacks agents from the central streamlines that exit the chamber comprises removing flow through one or more second fluid ports.

25. The method of claim 20, wherein removing flow from about the central streamlines to separate fluid flow that lacks agents from the central streamlines that exit the chamber comprises removing flow through one or more third fluid ports that are positioned between the first fluid port and one or more second fluid ports.

26. The method of claim 25, wherein flow through the one or more second fluid ports is prevented when reversing flow through the first fluid port.

27. The method of claim 25, wherein flow is introduced through the one or more second fluid ports when reversing flow through the first fluid port.