Multi-sheath flow and on-chip terminating electrode for microfluidic direct-blotting
Devices and methods are provided for the separation and dispensing of material using a microfluidic separation column connected via an exit channel to one or more sheath flow channels. The flow of separated material through the separation column is at least partially driven by a voltage potential between a first electrode within the separation column and a terminating electrode within at least one of the sheath flow channels. The separation column, exit channel, sheath flow channels, and electrodes are all within a single monolithic chip. The presence of an on-chip terminating electrode allows for separated material to be entrained in the sheath fluids and ejected onto a surface that can be non-conductive. The presence of multiple sheath flows allows for sheath flow fluids to have different compositions from one another, while reducing the occurrence of sheath flow fluids entering the separation column.
1. An analyte separation and dispensing apparatus comprising:
a separation column having an input end and an output end, wherein the input end has an opening configured to accept a fluid sample;
a first electrode within the separation column and proximate to the input end of the separation column;
an exit channel having an upstream end and a downstream end, wherein the upstream end of the exit channel is connected to the output end of the separation column, and wherein the downstream end of the exit channel has a discharge outlet;
a primary sheath fluid reservoir;
a pressure source connected to the primary sheath fluid reservoir;
a primary sheath flow channel, wherein an intersecting end of the primary sheath flow channel intersects the exit channel, and wherein the primary sheath flow channel connects the primary sheath fluid reservoir with the exit channel;
a secondary sheath flow channel, wherein an intersecting end of the secondary sheath flow channel intersects the primary sheath flow channel; and
a second electrode within the primary or secondary sheath flow channel.
2. The apparatus of claim 1 wherein the second electrode is within the primary sheath flow channel.
3. The apparatus of claim 1 wherein the second electrode is within the secondary sheath flow channel.
4. The apparatus of claim 1 further comprising:
a surface positioned across a gap from the discharge outlet; and
a motor configured to move the surface or the discharge outlet laterally with respect to one another.
5. The apparatus of claim 4 further comprising:
a membrane affixed to the surface, wherein a region of the membrane immediately across from the discharge outlet is dry until wetted by an effluent exiting from the discharge outlet, wherein the effluent comprises an analyte, thereby wicking the effluent into the membrane to immobilize the analyte in the membrane.
6. The apparatus of claim 5 further comprising:
a vacuum manifold connected to the surface, wherein the vacuum manifold is configured to affix the membrane to the surface and to immobilize the analyte in the membrane.
7. An analyte separation and dispensing apparatus comprising:
a separation column having an input end and an output end, wherein the input end has an opening configured to accept a fluid sample;
a first electrode within the separation column and proximate to the input end of the separation column;
an exit channel having an upstream end and a downstream end, wherein the upstream end of the exit channel is connected to the output end of the separation column, and wherein the downstream end of the exit channel has a discharge outlet;
a primary sheath fluid reservoir;
a first pressure source connected to the primary sheath fluid reservoir;
a primary sheath flow channel, wherein an intersecting end of the primary sheath flow channel intersects the exit channel, and wherein the primary sheath flow channel connects the primary sheath fluid reservoir with the exit channel;
a secondary sheath flow channel, wherein an intersecting end of the secondary sheath flow channel intersects (1) the primary sheath flow channel, or (2) the exit channel at an exit channel location between the exit channel discharge outlet and the intersection of the primary sheath flow channel and the exit channel;
an organic solvent reservoir;
a second pressure source connected to the organic solvent reservoir,
a tertiary sheath flow channel connecting the organic solvent reservoir with the exit channel,
wherein an intersecting end of the tertiary sheath flow channel intersects the exit channel at an exit channel location between (1A) the exit channel discharge outlet and (1B) the intersection of the primary sheath flow channel and the exit channel, when the intersecting end of the secondary sheath flow channel intersects the primary sheath flow channel;
and wherein the intersecting end of the tertiary sheath flow channel intersects the exit channel at an exit channel location between (2A) the exit channel discharge outlet and (2B) the intersection of the secondary sheath flow channel and the exit channel, when the intersecting end of the secondary sheath flow channel intersects the exit channel at an exit channel location between the exit channel discharge outlet and the intersection of the primary sheath flow channel and the exit channel; and
a second electrode within the primary, secondary, or tertiary sheath flow channel.
8. The apparatus of claim 7 further comprising:
a surface positioned across a gap from the discharge outlet;
a motor configured to move the surface or the discharge outlet laterally with respect to one another; and
a third electrode connected to the surface, wherein the third electrode is configured to electrospray an effluent from the discharge outlet to the surface.
9. The apparatus of claim 1 further comprising:
a sieving gel, wherein the sieving gel is inside the separation column, the primary sheath fluid reservoir, and the primary sheath flow channel.
10. The apparatus of claim 9 further comprising:
a secondary sheath fluid, wherein the secondary sheath fluid is inside the secondary sheath flow channel.
11. The apparatus of claim 10 wherein one or both of the primary and secondary sheath fluids independently comprises one or more analyte modifiers.
12. The apparatus of claim 1 wherein the separation column and the flow channels are integrated on a single monolithic chip.
13. The apparatus of claim 12 wherein further comprising:
a chip-mounted seal at an entrance end of at least one of the flow channels.
14. The apparatus of claim 1 wherein the pressure source connected to the primary sheath fluid reservoir is a first pressure source, and wherein the apparatus further comprises:
a second pressure source connected to the secondary sheath flow channel.
15. The apparatus of claim 1 wherein the pressure source connected to the primary sheath fluid reservoir is a first pressure source, and wherein the apparatus further comprises:
an organic solvent reservoir;
a second pressure source connected to the organic solvent reservoir; and
a tertiary sheath flow channel connecting the organic solvent reservoir with the exit channel, wherein an intersecting end of the tertiary sheath flow channel intersects the exit channel at a location between the exit channel discharge outlet and the intersection of the primary sheath flow channel and the exit channel.
16. The apparatus of claim 1 wherein the pressure source connected to the primary sheath fluid reservoir is an inkjet impulsive pump.
17. The apparatus of claim 7 further comprising:
a sieving gel, wherein the sieving gel is inside the separation column, the primary sheath fluid reservoir, and the primary sheath flow channel.
18. The apparatus of claim 17 further comprising:
a secondary sheath fluid, wherein the secondary sheath fluid is inside the secondary sheath flow channel.
19. The apparatus of claim 18 wherein one or both of the primary and secondary sheath fluids independently comprises one or more analyte modifiers.
20. The apparatus of claim 7 wherein the separation column and the flow channels are integrated on a single monolithic chip.
21. The apparatus of claim 20 wherein further comprising:
a chip-mounted seal at an entrance end of at least one of the flow channels.
22. The apparatus of claim 7 wherein the apparatus further comprises:
a third pressure source connected to the secondary sheath flow channel.
23. The apparatus of claim 7 wherein the first pressure source connected to the primary sheath fluid reservoir is an inkjet impulsive pump.