Flow cell for synthesis of arrays of DNA probes and the like

• US 6,444,175 B1
• Filed: 11/09/2001
• Issued: 09/03/2002
• Est. Priority Date: 11/10/1999
• Status: Expired due to Term

• Current Assignee: Wisconsin Alumni Research Foundation (University of Wisconsin System)
• Original Assignee: Wisconsin Alumni Research Foundation (University of Wisconsin System)
• Inventors: Singh-Gasson, Sangeet, Green, Roland D., Yue, Yongjian
• Primary Examiner(s): Warden, Jill
• Assistant Examiner(s): Gakh, Yelena G
• Application Number:
  US10/053,368
• Publication Number:
  US 20020061262A1
• Time in Patent Office: 298 Days
• Field of Search: 422/50, 422/70, 422/99, 422/100-104, 435/6, 435/7.1, 356/246, 436/518
• US Class Current: 422/553
• CPC Class Codes: B01J 19/0046 Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays synthesis methods per se C40B50/00
• CPC Class Codes: B01J 2219/00281 Individual reactor vessels
• CPC Class Codes: B01J 2219/00351 Means for dispensing and evacuation of reagents
• CPC Class Codes: B01J 2219/00527 Sheets
• CPC Class Codes: B01J 2219/00596 Solid-phase processes
• CPC Class Codes: B01J 2219/00605 the compounds being directly bound or immobilised to solid supports
• CPC Class Codes: B01J 2219/00612 the surface being inorganic
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: C40B 40/06 Libraries containing nucleotides or polynucleotides, or derivatives thereof
• CPC Class Codes: C40B 60/14 for creating libraries
• CPC Class Codes: G01N 2035/00158 Elements containing microarrays, i.e. "biochip"
• CPC Class Codes: G01N 35/0098 involving analyte bound to insoluble magnetic carrier, e.g. using magnetic separation magnetic particles used in immunoassays G01N33/54326; magnetic separation in general B03C

Abstract

A flow cell which can be used in the synthesis of DNA probes on an active surface of the substrate includes a base having a central window opening and a registration surface against which the substrate may be mounted. A gasket having a central opening defining an active area is mounted on the active surface of the substrate and has inlet and output extension openings which extend away from the central opening. A press block is engaged against the gasket to fully enclose an active volume between the press block, the peripheral walls of the central opening in the gasket, and the active surface of the substrate. A press is mounted to the base to selectively press the press block against the gasket and hold it in position. Channels in the press block extend to the extension openings in the gasket to allow flow of reagent into and out of the active volume, which is confined to the thickness of the gasket and can thus be minimized. The press may include a press screw which can be turned to engage against the press block to hold it into position, and which can be turned to release the press block from the gasket, allowing rapid and easy replacement of substrates.

Claims

1. A flow cell of the type that may be used in the synthesis of arrays of DNA probes and the like, comprising:

(a) a transparent substrate with two parallel flat surfaces, one of the surfaces forming an active surface of the substrate;

(b) a base having a central window opening and a registration surface against which the substrate is mounted;

(c) a gasket mounted on the active surface of the substrate and having a central opening defining an active area surrounded by material of the gasket, an inlet extension opening extending from the central opening of the gasket, and an outlet extension opening extending from the central opening of the gasket; and

(d) a press block having an engagement surface which is engaged against the gasket to press the gasket against the substrate mounted on the base to form a sealed volume defined by the periphery of the openings in the gasket and the active surface of the substrate and the engagement surface of the press block.

2. The flow cell of claim 1 further including an inlet channel in the press block extending from an exterior surface thereof to communication with the inlet extension opening in the gasket and an outlet channel in the press block extending from communication with the outlet extension opening in the gasket to an external surface of the press block.

3. The flow cell of claim 1 wherein the registration surface of the base is raised above adjacent areas of the base and surrounds the central window opening in the base, the registration surface formed flat to allow the precise location of the active surface of the transparent substrate to be defined with respect to an optical image projected onto the substrate through the window opening of the base.

4. The flow cell of claim 3 further including a reference surface on the base that is parallel to the registration surface whereby the base can be mounted with the reference surface against a surface of an image former to thereby locate the parallel registration surface.

5. The flow cell of claim 1 wherein the gasket is formed of a thin, non-reactive material having parallel opposite flat surfaces.

6. The flow cell of claim 5 wherein the gasket thickness is less than one mm.

7. The flow cell of claim 5 wherein the total volume enclosed by the central opening of the gasket between the engagement surface and the substrate surface is less than 100 microliters.

8. The flow cell of claim 7 wherein the thickness of the gasket is about 0.25 mm.

9. The flow cell of claim 1 including a standing frame secured to the base having an upright section extending from the base and an arm section extending from the upright section over the window opening in the base, and a press screw threadingly engaged with the arm section with a drive end thereof positioned to engage against an external surface of the press block as the press screw is turned to thread it toward the press block.

10. The flow cell of claim 9 wherein the drive end of the press screw is rounded.

11. The flow cell of claim 10 wherein the external surface of the press block opposite to the engagement surface has a concave depression formed therein which is fitted to receive the rounded drive end of the press screw to form a ball-and-socket engagement.

12. A flow cell of the type that may be used in the synthesis of arrays of DNA probes and the like, comprising:

(a) a transparent substrate with two parallel flat surfaces, one of the surfaces forming an active surface of the substrate;

(b) a base having a central window opening and a registration surface against which the substrate is mounted;

(c) a press block having an engagement surface such that the substrate is engaged between the press block and the registration surface of the base, the press block having an external surface opposite to the engagement surface with a concave depression formed therein; and

(d) a press comprising a standing frame secured to the base and having an upright section extending from the base and an arm section extending from the upright section over the central window opening in the base, and a press screw threadingly engaged with the arm section and positioned to engage the concave depression in the external surface of the press block with a rounded drive end thereof as the press screw is turned to thread it toward the press block.

13. The flow cell of claim 12 further including a gasket having a central opening defining an active area surrounded by the material of the gasket, an input extension opening extending from the central opening and an output extension opening extending from the central opening, the gasket mounted on the substrate between the substrate active surface and the engagement surface of the press block.

14. The flow cell of claim 13 wherein the press block includes an inlet channel extending from an external surface to communication with the inlet extension opening in the gasket and an outlet channel in the press block extending from communication with the outlet extension opening in the gasket to an external surface of the press block.

15. The flow cell of claim 13 wherein the gasket is formed of a thin, non-reactive material having parallel opposite flat surfaces.

16. The flow cell of claim 15 wherein the gasket thickness is less than one mm.

17. The flow cell of claim 15 wherein the total volume enclosed by the central opening of the gasket between the engagement surface and the substrate surface is less than 100 microliters.

18. The flow cell of claim 17 wherein the thickness of the gasket is about 0.25 mm.

19. The flow cell of claim 12 wherein the registration surface of the base is raised above adjacent areas of the base and surrounds the central window opening in the base, the registration surface formed flat to allow the precise location of the active surface of a transparent substrate to be defined with respect to an optical image projected onto the substrate through the window opening of the base.

20. The flow cell of claim 19 further including a reference surface on the base that is parallel to the registration surface whereby the base can be mounted with the reference surface against a surface of an image former to thereby locate the parallel registration surface.