Membrane-integrated microfluidic device for imaging cells

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, and wherein a surface of at least one of the first chamber and the second chamber comprises a three-dimensional structure to induce fluid mixing;
• (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;
• (h) 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;
• 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;
• a height and a shape of the first manifold structure and the second manifold structure are selected to permit an objective lens of the microscope to come into close proximity to a surface of the microfluidic device proximate to the second polymeric layer and to permit a second objective lens of a second microscope to come into close proximity to a second surface of the microfluidic device proximate to the first transparent, solid support layer;
• 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;
• and
• 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.