Additive channels
Compositions, devices and methods are described for preventing, reducing, controlling or delaying adhesion, adsorption, surface-mediated clot formation, or coagulation in a microfluidic device or chip. In one embodiment, blood (or other fluid with blood components) that contains anticoagulant is introduced into a microfluidic device comprising one or more additive channels containing one or more reagents that will re-activate the native coagulation cascade in the blood that makes contact with it “on-chip” before moving into the experimental region of the chip.
1. A method of adding reagent that re-activates a coagulation cascade to a fluid sample in a microfluidic device, comprising the following steps in sequential order:
a) providing
i) a fluid sample comprising anticoagulant, and
ii) a microfluidic device comprising an additive channel and at least one microchannel comprising a region comprising cells, said cells are attached to a cell adhesion promoting substance that coats said microchannel, said additive channel in fluidic communication with said at least one microchannel, said additive channel comprising
iii) a solution without cells and comprising reagent that re-activates said coagulation cascade, and
b) introducing said fluid sample into said at least one microchannel of said microfluidic device such that a portion of said fluid sample contacts said solution as said fluid sample moves through said at least one microchannel.
2. The method of claim 1 , wherein said fluid sample comprises platelets.
3. The method of claim 2 , wherein said fluid sample is blood.
4. The method of claim 3 , wherein said blood is human blood.
5. The method of claim 4 , further comprising adding said anticoagulant to said human blood at the time of collection of said blood from said human.
6. The method of claim 1 , wherein said microfluidic device comprises an input port in fluidic communication with said at least one microchannel and said introducing of step b) is through said input port.
7. The method of claim 6 , wherein said additive channel is positioned at or near said input port.
8. The method of claim 7 , wherein said additive channel is a first additive channel positioned on one side of said at least one microchannel near said input port.
9. The method of claim 1 , wherein said additive channel is a second additive channel positioned on another side of said at least one microchannel near said input port.
10. The method of claim 1 , wherein said anticoagulant is sodium citrate.
11. The method of claim 1 , wherein said anticoagulant is ethylenediamine tetraacetic acid (EDTA).
12. The method of claim 1 , wherein said cells are living cells.
13. The method of claim 1 , wherein said cells are fixed cells.
14. The method of claim 1 , wherein said cells comprise endothelial cells.
15. The method of claim 14 , wherein said endothelial cells are vascular endothelial cells.
16. The method of claim 15 , wherein said vascular endothelial cells are a monolayer.
17. The method of claim 16 , wherein said monolayer is disposed on a membrane.
18. The method of claim 1 , wherein said cell adhesion promoting substance comprises one or more extracellular matrix (ECM) proteins.
19. A method of adding reagent to a fluid sample in a microfluidic device, comprising the following steps in sequential order:
a) providing
i) a fluid sample comprising anticoagulant, and
ii) a microfluidic device comprising an additive channel and at least one microchannel comprising a region comprising endothelial cells, said endothelial cells are attached to a cell adhesion promoting substance that coats said microchannel, said additive channel is in fluidic communication with said at least one microchannel, said additive channel comprising a solution without cells comprising reagent that re-activates a coagulation cascade; and
b) introducing said fluid sample into said at least one microchannel of said microfluidic device such that a portion of said fluid sample contacts said solution as said fluid sample moves through said at least one microchannel.
20. The method of claim 19 , wherein said endothelial cells are vascular endothelial cells.
21. The method of claim 20 , wherein said vascular endothelial cells are a monolayer.
22. The method of claim 21 , wherein said monolayer is disposed on a membrane.
23. The method of claim 19 , wherein said cell adhesion promoting substance comprises one or more extracellular matrix (ECM) proteins.