High throughput microfluidic device
The present invention provides microfluidic devices capable of generating a flow of fluids within a chamber. The flow of fluids mix within the chamber to form a gradient of concentrations. The chamber is capable of accepting a cell culture and is accessible to retrieve the cell culture. The microfluidic devices are compatible with standard laboratory microscopes for optical analyses of the chamber.
1 . A microfluidic device comprising:
a first layer having a top and bottom surface, a thickness in-between, and a plurality of wells and a test chamber embedded in the top surface, each of the wells being fluidly connected to the test chamber by at least one microchannel, and a vacuum port embedded in the top surface fluidly connected to a vacuum channel extending to a position near the test chamber;
a second layer having a top and bottom surface, a thickness in-between, and a plurality of well openings, a vacuum port opening and a test chamber opening extending between the top and bottom surface, the test chamber opening being in alignment with the test chamber of the first layer, and an indentation embedded in the top surface forming a perimeter around the test chamber opening, the indentation comprising an aperture that extends through to the bottom surface of the second layer;
wherein the second layer is attachable to the top surface of the first layer such that each of the well openings of the second layer align with each of the wells of the first layer, the vacuum port opening of the second layer aligns with the vacuum port of the first layer, and the aperture of the second layer aligns with the vacuum channel of the first layer, and wherein the first and second layers are releasably interfaced.
2 . The device of claim 1 , further comprising a coverslip sized to cover the indentation and the test chamber opening of the second layer.
3 . The device of claim 1 , further comprising a third layer having a top and bottom surface, a thickness in-between, a plurality of well openings extending between the top and bottom surface, and a window opening extending between the top and bottom surface, wherein the third layer is attachable to the top surface of the third layer such that each of the well openings of the third layer align with each of the well openings of the second layer, and the window opening surrounds the indentation of the second layer.
4 . The device of claim 1 , further comprising a support layer attached to the bottom surface of the first layer.
5 . The device of claim 1 , wherein the first layer and the second layer each have at least two alignment slots that extend through top and bottom surfaces of each layer, each alignment slot being sized to accept a guide rod.
6 . The device of claim 5 , wherein the alignment slots are positioned such that passing the alignment slots of the second layer over guide rods inserted into the alignment slots of the first layer aligns each well opening of the second layer with each well of the first layer.
7 . The device of claim 1 , further comprising one or more bubble traps or degassing valves connected to the microchannels, the test chamber, or both, wherein the one or more bubble traps comprise an inner diameter greater than an inner diameter of each of the at least one microchannel.
8 . The device of claim 1 , further comprising one or more diffusor regions fluidly connecting two or more of the at least one microchannel, wherein the one or more diffusor regions are configured to combine fluid from the two or more microchannels to form a gradient of one or more agents that flow to the test chamber.
9 . The device of claim 8 , wherein the one or more agents comprise RNA.
10 . The device of claim 9 , further comprising one or more bubble traps in the at least one microchannel upstream from the test chamber.
11 . A microfluidic device comprising:
a microchannel layer having a top surface, a bottom surface and a thickness in-between, the microchannel layer including:
a test chamber recessed into the top surface, the test chamber having a perimeter with a plurality of inlets and at least one outlet about the perimeter;
a plurality of inlet wells and at least one outlet well recessed into the top surface;
at least one vacuum port recessed into the top surface; and
a plurality of microchannels recessed into the top surface, wherein at least one microchannel fluidly connects each of the plurality of inlet wells to a corresponding test chamber inlet of the plurality of inlets about the perimeter of the test chamber, at least one microchannel fluidly connects the at least one outlet well to the test chamber outlet, and at least one vacuum microchannel having a first end and a second end, with the first end being fluidly connected to the vacuum port and fluidly isolated from the test chamber;
a cover layer having a top surface, a bottom surface and a thickness in-between, the bottom surface of the cover layer being positioned on, and in contact with, the top surface of the microchannel layer, the cover layer further including:
a plurality of well openings passing through the top and bottom surfaces and aligned with each of the plurality of inlet wells of the microchannel layer;
a test chamber opening having a perimeter and passing through the top and bottom surfaces and aligned with the test chamber of the microchannel layer;
an indentation recessed into the top surface and positioned around the perimeter of the test chamber opening, wherein the indentation includes an aperture passing through the thickness to the bottom surface of the cover layer and is fluidly connected to the second end of the vacuum microchannel; and
a coverslip having a perimeter edge, wherein the coverslip is sized such that the perimeter edge rests on the indentation and covers the test chamber opening and the indentation aperture.