Microfluidic flow assay and methods of use

Abstract

A method for evaluating a blood product of an individual are provided. Specifically, a method to utilize a microfluidic flow assay, which includes a substrate surface comprising lipid coated particles and microfluidic channels through which a blood product can flow. The lipid coated particles comprise functional molecules that can induce or inhibit the coagulation cascade.

What is claimed: 
     
         1 . A method for evaluating a blood product of an individual, comprising:
 a. perfusing the individual's blood product over a microfluidic device under flow conditions to contact the blood product with a functional molecule of a plurality of coated lipid particles, wherein the microfluidic device, comprises at least one microfluidic channel; and at least one substrate surface provided in the at least one microfluidic channel, wherein the at least one substrate surface comprises a plurality of lipid coated particles immobilized on the substrate surface, wherein the plurality of lipid coated particles comprises at least one functional molecule, wherein the at least one functional molecule induces coagulation; and   b. detecting one or more coagulation products associated with the at least one functional molecule of the plurality of the lipid coated particles.   
     
     
         2 . The method of  claim 1 , wherein the blood product is selected from the group consisting of whole blood, plasma, platelet rich plasma, and platelet poor plasma. 
     
     
         3 . The method of  claim 1 , wherein the flow conditions simulate hemodynamic conditions of the individual. 
     
     
         4 . The method of  claim 1 , wherein the functional molecule is one or more transmembrane proteins. 
     
     
         5 . The method of  claim 1 , wherein the transmembrane protein is selected from the group consisting of tissue factor, thromobomodulin, endothelial cell protein C receptor, glycoprotein Ilb/IIIa, glycoprotein VI, glycoprotein 1b/IX/V, P-selectin, glycoprotein IV, CD9, platelet endothelial cell adhesion molecule (PECAM-1), Ras-related protein 1b (rap1b), c-type lectin-like receptor 2 (CLEC-2), intracellular adhesion molecule 1 (ICAM-1), intracellular adhesion molecule 2 (ICAM-2) and combinations thereof. 
     
     
         6 . The method of  claim 1 , wherein the functional molecule initiates coagulation. 
     
     
         7 . The method of  claim 1 , wherein the functional molecule inhibits coagulation. 
     
     
         8 . The method of  claim 1 , wherein the step of detecting comprises quantifying the one or more coagulation products. 
     
     
         9 . The method of  claim 1 , wherein the one or more coagulation products consist of proteins selected from the group consisting of thrombin, fibrin, thrombin-antithrombin complex, fibrinopeptide A, fibrinopeptide B, D-dimer, prothrombin fragment 1+2, activated factor X, activated factor V, activated factor VIIIa, activated factor IXa, activated factor XIa, activated factor XIIa, activated protein C, activated protein S, and mixtures thereof. 
     
     
         10 . The method of  claim 1 , wherein the one or more coagulation products are detected by a method selected from the group consisting of brightfield microscopy, darkfield microscopy, fluorescence microscopy, multi-photon excitation, second harmonic generation, third harmonic generation, atomic force microscopy, scanning electron microscopy, and absorbance. 
     
     
         11 . The method of  claim 1 , wherein the plurality of the lipid coated particles comprises a plurality of particles having a hydrophilic surface. 
     
     
         12 . The method of  claim 1 , wherein the plurality of lipid coated particles comprises one or more phospholipid structures selected from the group consisting of phosphatidylserine, phosphatidylcholine, phosphatidic acid, phosphatidylethanolamine, phophoinositides, phosphosphingolipids, and combinations thereof. 
     
     
         13 . The method of  claim 1 , wherein the plurality of lipid coating particles are immobilized on the substrate surface by a method selected from covalent bonding, electrostatic interactions or hydrogen bonding. 
     
     
         14 . The method of  claim 1 , wherein the at least one microfluidic channel is capable of receiving fluid at a first end of the at least one microfluidic channel and allowing the fluid to flow through the at least one microfluidic channel to a second end of the at least one microfluidic channel. 
     
     
         15 . The method of  claim 1 , wherein the at least one microfluidic channel is split into multiple channels. 
     
     
         16 . The method of  claim 1 , wherein the at least one functional molecule is a tissue factor or a thrombomodulin. 
     
     
         17 . The method of  claim 1 , wherein a flow rate of the individual's blood product is between about 50 to 2600 sec −1 . 
     
     
         18 . The method of  claim 1 , wherein a flow rate of the individual's blood product is between 0 and about 500,000 sec −1 . 
     
     
         19 . The method of  claim 1 , further comprising an agent wherein the agent is an anticoagulant agent or coagulating agent. 
     
     
         20 . The method of  claim 1 , wherein the individual is a mammal.