Microfluidic flow assay for measuring hemostatic phenotypes
A microfluidic-based flow assay and methods of manufacturing the same are provided. Specifically, the microfluidic flow assay includes a micropatterned surface that induces clot formation and an array of microfluidic channels though which blood flows. The micropatterned surface contains two clotting stimuli, one for inducing platelet adhesion and another for inducing the coagulation cascade.
1. A microfluidic device, comprising:
at least one microfluidic channel; and
at least one prothrombotic surface provided in the at least one microfluidic channel, wherein the at least one prothrombotic surface comprises a cell adhesive protein and at least one structure, wherein the at least one structure consists essentially of a protein embedded in a lipid that induces coagulation, wherein the cell adhesive protein and the at least one structure are in distinct regions of the at least one prothrombotic surface.
2. The device of claim 1 , wherein the cell adhesive protein is selected from the group consisting of collagen and von Willebrand factor.
3. The device of claim 2 , wherein the cell adhesive protein is collagen.
4. The device of claim 2 , wherein the cell adhesive protein is von Willebrand factor.
5. The device of claim 1 , wherein the the protein embedded in the lipid is tissue factor.
6. The device of claim 5 , wherein the tissue factor is interspersed in a predetermined geometry throughout the cell adhesive protein.
7. The device of claim 6 , wherein a surface area of the at least one prothrombotic surface is at least ten times smaller than a surface area of the at least one microfluidic channel.
8. The device of claim 6 , wherein a surface area of the at least one prothrombotic surface is at least one hundred times smaller than a surface area of the at least one microfluidic channel.
9. The device of claim 5 , wherein the at least one structure has a diameter of between about 10 microns to about 100 microns.
10. The device of claim 5 , wherein the at least one structure has a diameter of about 10 microns.
11. The device of claim 1 , wherein the at least one structure are provided in the cell adhesive protein and wherein each of the at least one structure have a width of less than 100 microns.
12. The device of claim 1 , wherein the at least one prothrombotic surface comprises a surface area of less than 10,000 square microns.
13. The device of claim 1 , further comprising:
a first antithrombotic region, wherein the first antithrombotic region is in the at least one microfluidic channel and positioned in a region proceeding a clot forming region in the at least one prothrombotic surface; and
a second antithrombotic region, wherein the second antithrombotic region is in the at least one microfluidic channel and positioned in a region following the clot forming region in the at least one prothrombotic surface.
14. The device of claim 13 , wherein the first antithrombotic region and the second antithrombotic region comprise a passivity protein, wherein the passivity protein does not induce platelet adhesion or coagulation cascade.
15. The device of claim 14 , wherein the first antithrombotic region comprises bovine serum albumin.
16. The device of claim 1 , wherein the at least one prothrombotic surface has a cross sectional area of about 50 microns by about 250 microns.
17. The device of claim 1 , wherein the ratio of the cell adhesive protein and the at least one structure in the at least one prothrombic surface is less than 1:1.
18. A microfluidic device made by a method, the method comprising:
providing a substrate;
creating at least one prothrombotic surface on the substrate, wherein the at least one prothrombotic surface comprises a cell adhesive protein and at least one structure, wherein the at least one structure consists essentially of a protein embedded in a lipid that induces coagulation, wherein the cell adhesive protein and the at least one structure are in distinct regions of the at least one prothrombotic surface; and
establishing at least one microfluidic channel which intersects at least a portion of the at least one prothrombotic surface.
19. A microfluidic channel through which blood is capable of flowing, the channel comprising:
at least one prothrombotic surface provided as a part of at least a portion of one surface in the channel, wherein the at least one prothrombotic surface comprises a cell adhesive protein and at least one structure, wherein the at least one structure consists essentially of a protein embedded in a lipid that induces coagulation, wherein the cell adhesive protein and the at least one structure are in distinct regions of the at least one prothrombotic surface, wherein a surface area of the at least one prothrombotic surface is at least one hundred times smaller than a surface area of the at least one microfluidic channel, wherein the at least one structure are provided in the collagen and wherein each of the at least one structures have a surface area of less than 100 microns, wherein the protein embedded in the lipid is tissue factor, and wherein the cell adhesive protein is selected from the group consisting of collagen and von Willebrand factor.
20. A kit for measuring clotting characteristics of blood, the kit comprising:
a hermetically sealed microfluidic device, the microfluidic device comprising at least one microfluidic channel and at least one prothrombotic surface provided in the at least one microfluidic channel, wherein the at least one prothrombotic surface comprises a cell adhesive protein and at least one structure, wherein the at least one structure consists essentially of a protein embedded in a lipid that induces coagulation, wherein the cell adhesive protein and the at least one structure are in distinct regions of the at least one prothrombotic surface.