Methods and apparatus for the location and concentration of polar analytes using an alternating electric field
A method is disclosed for effecting the concentration of a polar analyte in an alternating electric field. In the method, a relative translation of the polar analyte and an alternating electric field along a translation path is effected. A portion of the polar analyte is then trapped and concentrated in a concentration zone formed by the intersection of the translation path and the alternating electric field. Also disclosed are various devices for carrying out the forgoing method.
1. An apparatus for concentrating analytes, comprising:
a first reservoir containing at least one electrode;
a second reservoir containing at least one electrode;
a channel in fluid communication with the first and second reservoirs, the channel including a concentration zone; and
at least two electrodes located on opposite sides of the channel and outside the first and second reservoirs, the at least two electrodes being configured to subject the concentration zone to an alternating electric field.
2. The apparatus of claim 1 , further comprising a power supply configured to provide a voltage difference between the at least one electrode contained in the first reservoir and the at least one electrode contained in the second reservoir.
3. The apparatus of claim 1 , further comprising a power supply configured to provide a time-variant voltage difference between the at least two electrodes that are located on opposite sides of the channel and outside the first and second reservoirs.
4. The apparatus of claim 1 , further comprising a hybridization array located in the concentration zone.
5. The apparatus of claim 4 , wherein the hybridization array is a polynucleotide hybridization array.
6. The apparatus of claim 1 , further comprising a frit including an insulating matrix having pores containing a fluid medium.
7. The apparatus of claim 6 , wherein the pores have an effective internal diameter of between about 0.5 μm and 50 μm, and the electrical conductivity of the fluid medium is between 10 and 1000 times greater than the electrical conductivity of the insulating matrix.
8. The apparatus of claim 6 , wherein the frit further includes electrically-conductive particles suspended in the insulating matrix.
9. The apparatus of claim 1 , wherein the at least two electrodes that are located on opposite sides of the channel and outside the first and second reservoirs are isolated from the channel and are spaced from one another by between about 50 μm and 2 mm.
10. The apparatus of claim 1 , wherein the at least two electrodes that are located on opposite sides of the channel and outside the first and second reservoirs have substantially identical surfaces facing one another, and the shape of the substantially identical surfaces is one of flat, pointed, and rounded.
11. A method for making an analyte concentrator, comprising:
providing, by etching, laser micromachining, or photolithography, a fluidic device including a first reservoir containing at least one electrode, a second reservoir containing at least one electrode, and a channel in fluid communication with the first and second reservoirs, the channel including a concentration zone; and
providing at least two electrodes on opposite sides of the channel and outside the first and second reservoirs, the at least two electrodes being configured to subject the concentration zone to an alternating electric field.
12. The method of claim 11 , further comprising:
connecting a first power supply to the at least one electrode contained in the first reservoir and the at least one electrode contained in the second reservoir, the first power supply being configured to provide a voltage difference between the at least one electrode contained in the first reservoir and the at least one electrode contained in the second reservoir; and
connecting a second power supply to the at least two electrodes that are located on opposite sides of the channel and outside the first and second reservoirs, the second power supply being configured to provide a time-variant voltage difference between the at least two electrodes that are located on opposite sides of the channel and outside the first and second reservoirs.
13. The method of claim 11 , further comprising arranging a polynucleotide hybridization array in the concentration zone.
14. The method of claim 11 , further comprising arranging a frit including an insulating matrix having pores containing a fluid medium in the concentration zone, the pores having an effective internal diameter of between about 0.5 μm and 50 μm, and the electrical conductivity of the fluid medium being between 10 and 1000 times greater than the electrical conductivity of the insulating matrix.
15. The method of claim 11 , further comprising spacing the at least two electrodes that are located on opposite sides of the channel and outside the first and second reservoirs by between about 50 μm and 2 mm, and isolating these electrodes from the channel.
16. A method for performing high-throughput concentration of analytes, comprising:
manipulating, with a robotic system with fluid handling capability, a plurality of microfluidic devices each including a first reservoir containing at least one electrode, a second reservoir containing at least one electrode, a channel in fluid communication with the first and second reservoirs, and at least two electrodes located on opposite sides of the channel and outside the first and second reservoirs, the at least two electrodes being configured to subject a first portion of the channel to an alternating electric field;
transferring, with the robotic system with fluid handling capability, a supporting medium in the first and second reservoirs and the channel of each of the plurality of microfluidic; and
introducing, with the robotic system with fluid handling capability, a polar analyte in the first reservoir of each of the plurality of microfluidic devices.
17. The method of claim 16 , further comprising:
subjecting the first portion of the channel in each of the plurality of microfluidic devices to an alternating electric field by providing a time-variant voltage difference between the at least two electrodes that are located on opposite sides of the channel and outside the first and second reservoirs; and
electrokinetically translating the polar analyte in each of the plurality of microfluidic devices from the first reservoir toward the second reservoir by providing a voltage difference between the at least one electrode contained in the first reservoir and the at least one electrode contained in the second reservoir.
18. The method of claim 17 , further comprising detecting the polar analyte in the first portion of the channel in each of the microfluidic devices that is subjected to an alternating electric field.
19. The method of claim 18 , wherein the polar analyte is a nucleic acid, and further comprising subjecting the nucleic acid in each of the microfluidic devices to a nucleic acid hybridization reaction in which the nucleic acid is contacted with at least one complementary nucleic acid.
20. The method of claim 19 , wherein the at least one complementary nucleic acid to which the nucleic acid is contacted in each of the microfluidic devices is a synthetic polynucleotide probe or a cDNA molecule.