Microfluidic flow cell assemblies for imaging and method of use
A microfluidic flow cell subassembly, which may be assembled into a flow cell having fluidic connections outside of the main substrate, is described for encapsulating a sample to allow for subsequent controlled delivery of reagents to the sample, such as multiplexed in situ biomarker staining and analysis. As configured, the subassembly comprises a substrate layer forms a flexible optically transparent lid which is capable of bending in either direction to alter the internal dimensions of the subassembly. Methods of use are also disclosed.
1. A microfluidic subassembly comprising:
a stacked planar assembly comprising;
an adherent layer;
a transparent substrate layer;
a gasket layer; and
wherein each layer is adhered to one another and the adherent layer and the gasket layer extend beyond the extents of the substrate layer; and
at least one fluidic port wherein said port is positioned outside the boundaries of the substrate layer; and
where the substrate layer forms a flexible transparent lid capable of bending in either direction to alter the internal dimensions of the subassembly.
2. The subassembly of claim 1 where the transparent substrate layer comprises glass or plastic or a combination thereof.
3. The subassembly of claim 2 where the flexible transparent lid is configured to deflect in a range of plus or minus approximately 200 μm from its center point.
4. The subassembly of claim 3 wherein the flexible transparent lid is configured to deflect in a range of approximately −50 to +100 μm.
5. The subassembly of claim 3 wherein the flexible transparent lid is configured to deflect in a range of approximately −20 to +50 μm.
6. The subassembly of claim 1 where the internal dimensions has a volume capacity in the range of 1 μL to 1000 μL.
7. The subassembly of claim 6 wherein the volume capacity is in the range of 25 μL to 250 μL.
8. The subassembly of claim 1 wherein the fluidic port is a thin film fluidic connector where said connector comprises at least one microfluidic channel in fluid connection with the stacked planar assembly and positioned outside the boundaries of the substrate layer.
9. The subassembly of claim 8 wherein the thin film fluidic connector is connected to the gasket layer.
10. A microfluidic flow cell comprising:
a microfluidic subassembly comprising:
a stacked planar assembly comprising;
an adherent layer;
a transparent substrate layer; and
a gasket layer;
where each layer is adhered to one another and configured such that the adherent layer and the gasket layer extend beyond the extents of the substrate layer;
at least one fluidic port wherein said port is positioned outside the boundaries of the substrate layer; and
a solid support adhered to the microfluidic flow cell subassembly;
where the substrate layer forms a flexible transparent lid configured to bend towards or away from the solid support by applying negative or positive pressure respectively.
11. The flow cell of claim 10 where the transparent substrate layer comprises glass or plastic or a combination thereof.
12. The flow cell of claim 11 where the flexible transparent lid is configured to deflect in a range of plus or minus approximately 200 μm from its center point.
13. The flow cell of claim 12 wherein the flexible transparent lid is configured to deflect in a range of approximately −50 to +100 μm.
14. The flow cell of claim 12 wherein the flexible transparent lid is configured to deflect in a range of approximately −20 to +50 μm.
15. The flow cell of claim 11 where the internal dimensions has a volume capacity in the range of 1 μL to 1000 μL.
16. The flow cell of claim 15 wherein the volume capacity is in the range of 25 μL to 250 μL.
17. The flow cell of claim 11 wherein the fluidic port is a thin film fluidic connector where said connector comprises at least one microfluidic channel in fluid connection with the stacked planar assembly and is positioned outside the boundaries of the substrate layer.
18. The flow cell of claim 17 wherein the thin film fluidic connector is connected to the gasket layer.
19. The flow cell of claim 17 were the fluidic port further comprises a valve to control fluid flow and pressure in the microfluidic flow cell and is capable of connecting to a fluid delivery system.
20. The flow cell of claim 11 further comprising at least one attachment point configured to match attachment points of an imaging device.
21. The flow cell of claim 20 wherein the at least one attachment point is configured to align the flow cell with an objective lens of the imaging device.
22. A method of modulating the quality of an image captured of a sample supported on the solid support of the flow cell of claim 21 comprising the steps of:
aligning the flow cell with an objective lens of an imaging device; and
controlling the amount of liquid in the flow cell to deflect the flexible lid in a range of plus or minus approximately 200 μm from its center point.
23. The method of claim 22 where the flexible transparent lid is configured to deflect in a range of approximately −50 to +100 μm.
24. The method of claim 23 where the flexible transparent lid is configured to deflect in a range of approximately −20 to +50 μm.
25. The method of claim 22 where the sample is a biological sample.
26. The method of claim 22 further comprising the steps of capturing an image of the sample using an image capture device.