Membrane-integrated microfluidic device for imaging cells

Patent metadata

Field	Value
Publication number	US-9844779-B2
Application number	US-201213349830-A
Country	US
Kind code	B2
Filing date	Jan 13, 2012
Priority date	Jan 14, 2011
Publication date	Dec 19, 2017
Grant date	Dec 19, 2017

Abstract

A microfluidic device is provided. The microfluidic device includes a first transparent, solid support layer. A first polymeric layer defining at least one chamber is attached to the first transparent, solid support layer. A semi-permeable membrane is attached to the first polymeric layer. A second polymeric layer is attached to the opposite side of the semi-permeable membrane from the first polymeric layer. The second polymeric layer has a thickness of less than 300 microns and defines at least one chamber positioned to overlap with at least one chamber in the first polymeric layer. A first manifold structure is attached to an input end of at least one chamber and a second manifold structure is attached to an output end of at least one chamber.

First claim

We claim:

1. A microfluidic device comprising:
(a) a first transparent, solid support layer comprising a first optically clear material;
(b) a first polymeric layer comprising a second material different from the first material of the first transparent, solid support layer, wherein the first polymeric layer has a thickness of less than 200 μm, defines therein at least two chambers including a first chamber, and is attached to said first transparent, solid support layer;
(c) a semi-permeable membrane attached to the first polymeric layer, wherein at least a portion of a surface of the semi-permeable membrane is plasma treated;
(d) a second polymeric layer attached to the opposite side of the semi-permeable membrane from the first polymeric layer; wherein the second polymeric layer has a thickness of less than 300 μm, and defines therein at least two chambers including a second chamber that is positioned to overlap, at least partially, with at least the first chamber in the first polymeric layer, and wherein a cell culture is adhered to a surface of the semi-permeable membrane;
(e) a first manifold structure attached to an input end of at least one of the first and second chambers;
(f) a second manifold structure attached to an output end of at least one of the first and second chambers, wherein the first manifold structure and the second manifold structure are separated by a distance greater than about 20 millimeters; and
(g) a solid frame, wherein the first transparent, solid support layer, the first polymeric layer, the semi-permeable membrane, and the second polymeric layer are positioned within the solid frame such that the solid frame surrounds the edges of the microfluidic device, thereby leaving an upper surface and a lower surface of the microfluidic device clear for viewing using a microscope;
wherein a thickness of the second polymeric layer is selected to provide a predetermined level of oxygen gas permeance, structural rigidity, and an ability to observe the cell culture using a microscope positioned over the second polymeric layer and in alignment with at least one of the at least two chambers of the second polymeric layer.

2. The device of claim 1 , wherein the first transparent, solid support layer has a thickness in the range about 100 μm to about 170 μm.

3. The device of claim 1 , wherein the first transparent, solid support layer is made of glass, polyester, polycarbonate, polystyrene, poly(dimethylsiloxane), poly(methyl methacrylate), polyethylene, polyethylene terephthalate, polyurethane, polycaprolactone, polylactic acid, polyglycolic acid, poly(lactic-co-glycolic acid), polyglycerol sebacate, polyester amide, polydioxanone, cyclic olefin copolymer, quartz, or a transparent thermoplastic.

4. The device of claim 1 , wherein the first transparent, solid support layer is made of glass.

5. The device of claim 1 , wherein the first polymeric layer has a thickness in the range about 10 μm to about 130 μm.

6. The device of claim 1 , wherein the first polymeric layer is made of poly(dimethylsiloxane), polycarbonate, polystyrene, poly(methyl methacrylate), polyethylene, polyethylene terephthalate, polyurethane, polycaprolactone, polylactic acid, polyglycolic acid, poly(lactic-co-glycolic acid), polyglycerol sebacate, polyester amide, polydioxanone, cyclic olefin copolymer, polyimide, or polyamide.

7. The device of claim 1 , wherein the first polymeric layer is made of poly(dimethylsiloxane).

8. The device of claim 1 , wherein the semi-permeable membrane has a thickness of about 1 μm to about 20 μm.

9. The device of claim 1 , wherein the semi-permeable membrane is made of polycarbonate, poly(dimethylsiloxane), polyethersulfone, polysulfone, polystyrene, or polyester.

10. The device of claim 1 , wherein the semi-permeable membrane is made of polycarbonate.

11. The device of claim 1 , wherein the second polymeric layer has a thickness in the range about 150 μm to about 250 μm.

12. The device of claim 1 , wherein the second polymeric layer is made of poly(dimethylsiloxane), polycarbonate, polystyrene, poly(methyl methacrylate), polyethylene, polyethylene terephthalate, polyurethane, polycaprolactone, polylactic acid, polyglycolic acid, poly(lactic-co-glycolic acid), polyglycerol sebacate, polyester amide, polydioxanone, cyclic olefin copolymer, polyimide, or polyamide.

13. The device of claim 1 , wherein the second polymeric layer is made of poly(dimethylsiloxane).

14. The device of claim 1 , wherein any of the at least two chambers in the first polymeric layer has a height spanning from the first transparent, solid support layer to the semi-permeable membrane.

15. The device of claim 1 , wherein any of the at least two chambers in the first polymeric layer has a width in the range of from about 10 μm to about 1 cm.

16. The device of claim 1 , wherein any of the at least two chambers in the first polymeric layer has a width in the range of from about 100 μm to about 500 μm.

17. The device of claim 1 , wherein any of the at least two chambers in the second polymeric layer has a width in the range of from about 10 μm to about 1 cm.

18. The device of claim 1 , wherein any of the at least two chambers in the second polymeric layer has a width in the range of from about 100 μm to about 500 μm.

19. The device of claim 1 , wherein any of the at least two chambers in the second polymeric layer has a height in the range of from about 10 μm to about 150 μm.

20. The device of claim 1 , further comprising a second transparent, solid support layer attached to an opposite side of the second polymeric layer from the semi-permeable membrane.

21. The device of claim 20 , wherein the second transparent, solid support layer is made of glass, polyester, polycarbonate, polystyrene, poly(dimethylsiloxane), poly(methyl methacrylate), polyethylene, polyethylene terephthalate, polyurethane, polycaprolactone, polylactic acid, polyglycolic acid, poly(lactic-co-glycolic acid), polyglycerol sebacate, polyester amide, polydioxanone, cyclic olefin copolymer, quartz, or a transparent thermoplastic.

22. A method of viewing cells, comprising:
(a) providing a microfluidic device of claim 1 ;
(b) positioning a microscope to view cells attached to the semi-permeable membrane; and
(c) viewing said cells with said microscope.

23. The microfluidic device of claim 1 , wherein a surface of at least one of the first chamber and the second chamber comprises a three-dimensional structure to induce fluid mixing.

24. The microfluidic device of claim 1 , wherein at least one of the first chamber and the second chamber has a rounded cross sectional geometry selected to reduce shear stress experienced by a fluid traveling through the at least one of the first chamber and the second chamber.

25. The microfluidic device of claim 1 , wherein a surface of at least one of the first chamber and the second chamber is modified with at least one of a hydrophobic agent, a hydrophilic agent, a collagen, and an antibody.

26. The microfluidic device of claim 1 , further comprising a reservoir for fluid storage, the reservoir containing nutrients for the cell culture, wherein the reservoir comprises an extension of at least one of the first chamber and the second chamber.

27. The microfluidic device of claim 1 , wherein a height and a shape of the first manifold structure and the second manifold structure are selected to permit an

Assignees

• Borenstein Jeffrey T
• Charest Joseph L
• Cuiffi Joseph
• Epshteyn Alla
• Holton Angela B
• Hsiao James Ching-Ming
• Charles Stark Draper Laboratory Inc

Inventors

• Borenstein Jeffrey T
• Charest Joseph L
• Cuiffi Joseph
• Epshteyn Alla
• Holton Angela B
• Hsiao James Ching-Ming

Classifications

• G02B21/33

  Immersion oils {, or microscope systems or objectives for use with immersion fluids} · CPC title

• B01L2300/0887

  Laminated structure · CPC title

• B01L2300/0816

  Cards, e.g. flat sample carriers usually with flow in two horizontal directions · CPC title

• B01L3/502707 Primary

  characterised by the manufacture of the container or its components · CPC title

• B01L2200/0689

  Sealing · CPC title