Device for immobilizing chemical and biochemical species and methods of using same

• US 20060172408A1
• Filed: 12/01/2004
• Published: 08/03/2006
• Est. Priority Date: 12/01/2003
• Status: Abandoned Application

• Current Assignee: California Institute of Technology
• Original Assignee: California Institute of Technology
• Inventors: Quake, Steven R., Kartalov, Emil
• Application Number:
  US11/002,739
• Publication Number:
  US 20060172408A1
• Time in Patent Office: Days
• US Class Current: 435/287.200
• 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 2219/00432 Photolithographic masks
• CPC Class Codes: B01J 2219/00497 Features relating to the solid phase supports
• CPC Class Codes: B01J 2219/00527 Sheets
• CPC Class Codes: B01J 2219/00576 fluorophore
• CPC Class Codes: B01J 2219/00585 Parallel processes
• 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 2219/0063 Other, e.g. van der Waals forces, hydrogen bonding
• CPC Class Codes: B01J 2219/00637 by coating it with another layer
• CPC Class Codes: B01J 2219/00657 One-dimensional arrays
• CPC Class Codes: B01J 2219/00677 Ex-situ synthesis followed by deposition on the substrate
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: B01L 2200/12 Specific details about manufacturing devices
• CPC Class Codes: B01L 2300/0636 Integrated biosensor, microarrays
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0819 Microarrays; Biochips
• CPC Class Codes: B01L 2300/0864 comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
• CPC Class Codes: B01L 2300/0867 Multiple inlets and one sample wells, e.g. mixing, dilution
• CPC Class Codes: B01L 2300/0887 : Laminated structure
• CPC Class Codes: B01L 2400/0481 : squeezing of channels or chambers
• CPC Class Codes: B01L 2400/0688 : surface tension valves, capillary stop, capillary break
• CPC Class Codes: B01L 3/5025 : for parallel transport of multiple samples
• CPC Class Codes: B01L 3/502707 : characterised by the manufacture of the container or its components by shaping or joining plastic parts B29C59/00, B29C65/00; by laminating B32B37/00; manufacture of microstructural devices in general B81C

Abstract

A substrate is provided that facilitates immobilization of nucleic acid molecules, including DNA molecules and RNA molecules. Also provided is a device that includes the substrate, which, for example, can be a chip. In addition, methods of using the substrate are provided, including, for example, methods of sequencing a DNA molecule anchored to the substrate, and methods for conducting the process of sequencing using such devices.

Claims

1. A device, comprising a multi-layer polymeric structure, which comprises:

(a) a graft-copolymer comprising diacrylated polyglycol grafted to a second polymer; and

(b) a plurality of layers of a polyelectrolyte disposed over the graft-copolymer.

2. The device of claim 1, wherein the device is disposed on an optically transparent substrate.

3. The device of claim 2, wherein the device is a microfluidic device.

4. The device of claim 3, wherein the microfluidic device further comprises:

(a) a first layer defining a plurality of first channels;

(b) a second layer bonded to the first layer; and

(c) a sample of a DNA anchored to the inner surface of the first channels,

wherein the inner surface of each first channel is modified to include the multi-layer polymeric structure.

5. The device of claim 1, wherein the multi-layer polymeric structure further comprises biotin conjugated to the layer of the polyelectrolyte.

6. The device of claim 5, further comprising streptavidin disposed over biotin.

7. The device of claim 1, wherein the second polymer is selected from a group consisting of poly(dimethylsiloxane), poly(methylmethacrylate), and poly(urethane).

8. The device of claim 1, wherein the polyelectrolyte comprises alternating layers of a positively charged polymer and a negatively charged polymer.

9. The device of claim 8, wherein the positively charged polymer is selected from poly(ethyleneimine) and poly(allylamine).

10. The device of claim 8, wherein the negatively charged polymer is poly(acrylic acid).

11. The device of claim 2, wherein the optically transparent substrate comprises glass.

12. The device of claim 2, wherein the device is bonded to the optically transparent substrate.

13. The device of claim 12, wherein the device is bonded with poly(dimethylsiloxane), poly(methylmethacrylate), poly(urethane), or a combination thereof.

14. The microfluidic device of claim 4, wherein each first channel has a height of about 10 micrometers.

15. The microfluidic device of claim 4, wherein each first channel has the width of between about 60 and 150 micrometers.

16. The microfluidic device of claim 4, wherein the plurality of the first channels is between 2 and 50.

17. The microfluidic device of claim 4, wherein the second layer further defines a plurality of second channels.

18. The microfluidic device of claim 17, wherein each second channel has a height of about 10 micrometers.

19. The microfluidic device of claim 17, wherein each second channel has a width of between about 60 and 150 micrometers.

20. The microfluidic device of claim 17, wherein the plurality of the second channels is between 2 and 50.

29. An assembly, comprising a plurality of devices of claim 1.

30. The device of claim 1, wherein the polyglycol is poly(ethylene glycol).

21. A method for fabricating a microfluidic device, comprising:

(a) fabricating a first polymeric structure and a second polymeric structure, each structure defining a plurality of channels;

(b) aligning the first polymeric structure and the second polymeric structure,

wherein the channels in the first polymeric structure are not fluidically connected to the channels in the second polymeric structure, and

wherein the channels in the first polymeric structure and the channels in the second polymeric structure create a valve action;

(c) bonding the first polymeric structure to the second polymeric structure to obtain a fused structure;

(d) bonding the fused structure to an optically transparent substrate; and

(e) modifying the channels in the first polymeric structure by grafting a modifying polymer to the polymer forming the first polymeric structure;

to obtain a graft copolymer, and forming a layer of a polyelectrolytes over the graft copolymer,

thereby fabricating the microfluidic device.

22. The method of claim 21, wherein the modifying polymer is diacrylated poly(ethylene glycol).

23. The method of claim 21, further comprising applying a biotin over the layer of the polyelectrolytes.

24. The method of claim 23, further comprising applying streptavidin over biotin.

25. The method of claim 21, wherein forming the layer of the polyelectrolyte comprises applying layers of a positively charged polymer and a negatively charged polymer in alternating manner.

26. The method of claim 25, wherein the positively charged polymer is selected from poly(ethyleneimine) and poly(allylamine).

27. The method of claim 25, wherein the negatively charged polymer is poly(acrylic acid).

28. The method of claim 21, wherein the polymer forming the first polymeric structure and the second polymeric structure is poly(dimethylsiloxane), poly(methylmethacrylate), poly(urethane), or a combination thereof.

31. The method of claim 21, wherein the modifying polymer is diacrylated polyglycol.