Microfluidic devices, and methods of making and using the same
The present disclosure provides methods and systems for assaying a sample. A microfluidic device to perform an assay of a sample (e.g., biological sample) is described having a sample application site, a porous component and a flow channel. The porous component provides for uniform dissolution of a reagent and mixing of the sample and reagent without filtering the sample.
1. A microfluidic device comprising:
a sample application site;
a flow channel in fluid communication with the sample application site; and
a porous component positioned between the sample application site and flow channel, wherein the porous component comprises:
a porous matrix comprising pores; and
unbound dry assay reagent positioned within the pores of the porous matrix,
wherein the porous matrix has a length sufficient for the unbound dry assay reagent and a sample to substantially mix, and
wherein the porous matrix is configured to uniformly mix the assay reagent into a sample mixture flowing therethrough upon exit from the porous component and into the flow channel.
2. The microfluidic device according to claim 1 , wherein the porous matrix is configured to be non-filtering with respect to the sample for which the device is configured to assay.
3. The microfluidic device according to claim 1 , wherein the porous matrix comprises pores having diameters between 1 μm and 200 μm.
4. The microfluidic device according to claim 1 , wherein the porous matrix comprises a pore volume between 1 μL and 25 μL.
5. The microfluidic device according to claim 1 , wherein the porous matrix is a frit.
6. The microfluidic device according to claim 1 , wherein the porous component further comprises a buffer.
7. The microfluidic device according to claim 6 , wherein the buffer comprises bovine serum albumin (BSA), trehalose, polyvinylpyrrolidone (PVP) or 2-(N-morpholino) ethanesulfonic acid or a combination thereof.
8. The microfluidic device according to claim 7 , wherein the buffer comprises BSA, trehalose and PVP.
9. The microfluidic device according to claim 1 , wherein the assay reagent comprises an analyte-specific binding member.
10. The microfluidic device according to claim 9 , wherein the analyte-specific binding member comprises an antibody or analyte binding fragment thereof.
11. The microfluidic device according to claim 9 , wherein the analyte specific binding member is coupled to a detectable label.
12. The microfluidic device according to claim 11 , wherein the detectable label comprises a fluorescent dye.
13. The microfluidic device according to claim 1 , wherein the assay reagent comprises a chelating agent.
14. The microfluidic device according to claim 1 , wherein the flow channel comprises an optically transmissive wall.
15. The microfluidic device according to claim 14 , wherein the walls of the flow channel are optically transmissive to one or more of ultraviolet light, visible light and near-infrared light.
16. The microfluidic device according to claim 1 , where the sample application site is configured to receive a sample having a volume ranging from 5 μL to 2000 μL.
17. The microfluidic device according to claim 1 , wherein the device is configured to be hand held.
18. The microfluidic device according to claim 1 , wherein the porous matrix comprises a porous organic polymer.
19. A kit comprising:
a microfluidic device comprising:
a sample application site;
a flow channel in fluid communication with the sample application site; and
a porous component positioned between the sample application site and flow channel, wherein the porous component comprises a porous matrix comprising pores and unbound dry assay reagent positioned within the pores of the porous matrix, wherein the porous matrix has a length sufficient for the unbound dry assay reagent and a sample to substantially mix, and wherein the porous matrix is configured to uniformly mix the assay reagent into a sample mixture flowing therethrough upon exit from the porous component and into the flow channel; and
a container housing the device.