Integrated system for processing microfluidic samples, and method of using the same
This patent application describes an integrated apparatus for processing polynucleotide-containing samples, and for providing a diagnostic result thereon. The apparatus is configured to receive a microfluidic cartridge that contains reagents and a network for processing a sample. Also described are methods of using the apparatus.
1. An apparatus for processing and amplifying a plurality of polynucleotide-containing samples, the apparatus comprising:
a multi-lane microfluidic cartridge configured to process the plurality of polynucleotide-containing samples independently of each other, the multi-lane microfluidic cartridge comprising a plurality of sample lanes, each sample lane of the plurality of sample lanes comprising a dedicated pipette inlet and an amplification reaction zone, the multi-lane microfluidic cartridge further configured to permit the plurality of polynucleotide-containing samples to be loaded into the multi-lane microfluidic cartridge at different times, wherein the plurality of polynucleotide-containing samples are passed to the amplification reaction zone independently of one another, a sample lane of the multi-lane microfluidic cartridge further comprising a processing chamber;
a receiving bay configured to receive the multi-lane microfluidic cartridge;
a first heat source separate from the multi-lane microfluidic cartridge and configured to apply heat to the multi-lane microfluidic cartridge when the multi-lane microfluidic cartridge is received in the receiving bay, the first heat source configured to apply the heat to one or more selected regions of the multi-lane microfluidic cartridge at one or more selected times, in order to prepare one or more polynucleotides from the plurality of polynucleotide-containing samples for amplification, the first heat source configured to apply the heat to the processing chamber when the multi-lane microfluidic cartridge is received in the receiving bay;
a magnet configured to move into and out of place to apply a magnetic field to a polynucleotide-loaded retention member in the processing chamber of the multi-lane microfluidic cartridge when the multi-lane microfluidic cartridge is received in the receiving bay and the polynucleotide-loaded retention member is received in the multi-lane microfluidic cartridge;
a plurality of second heat sources separate from the multi-lane microfluidic cartridge and configured to align with and apply heat to one or more selected regions of the multi-lane microfluidic cartridge when the multi-lane microfluidic cartridge is received in the receiving bay, the plurality of second heat sources configured to apply the heat to the one or more selected regions of the multi-lane microfluidic cartridge at one or more selected times, in order to amplify the prepared one or more polynucleotides,
wherein a second heat source of the plurality of second heat sources is configured to cyclically heat in a series of heating phases after the multi-lane microfluidic cartridge is received in the receiving bay, wherein each heating phase comprises the second heat source being cycled between at least two temperatures,
wherein the second heat source of the plurality of second heat sources is configured to maintain a substantially uniform temperature in an amplification reaction zone at each temperature of the at least two temperatures when the multi-lane microfluidic cartridge is received in the receiving bay, and
wherein the plurality of second heat sources are configured to perform independent amplification reactions in the plurality of sample lanes when the multi-lane microfluidic cartridge is received in the receiving bay; and
an optical detector configured to detect the presence of the one or more amplified polynucleotides in the amplification reaction zone.
2. The apparatus of claim 1 , wherein the plurality of sample lanes are configured to share a wash buffer received in the multi-lane microfluidic from a reagent reservoir.
3. The apparatus of claim 1 , wherein the plurality of sample lanes are configured to share a release reagent received in the multi-lane microfluidic cartridge from a reagent reservoir.
4. The apparatus of claim 1 , wherein the plurality of sample lanes are configured to share a hydroxide solution received in the multi-lane microfluidic from a reagent reservoir.
5. The apparatus of claim 1 , further comprising the multi-lane microfluidic cartridge received in the receiving bay and configured to receive the heat applied by the first heat source and receive the heat applied by the plurality of second heat sources.
6. The apparatus of claim 1 , wherein the optical detector does not cover the dedicated pipette inlets of the multi-lane microfluidic cartridge when the multi-lane microfluidic cartridge is received in the receiving bay.
7. The apparatus of claim 1 , wherein the first heat source is configured to release one or more polynucleotides from the polynucleotide-loaded retention member in contact with a release reagent in the processing chamber when the multi-lane microfluidic cartridge is received in the receiving bay and the polynucleotide-loaded retention member is received in the processing chamber.
8. The apparatus of claim 1 , wherein each of the plurality of second heat sources comprises a plurality of resistive heaters.
9. The apparatus of claim 8 , wherein the plurality of resistive heaters is configured to maintain a substantially uniform temperature at each temperature of the at least two temperatures when the multi-lane microfluidic cartridge is received in the receiving bay.
10. The apparatus of claim 1 , wherein each of the plurality of second heat sources is fixed in position relative to the multi-lane microfluidic cartridge during cyclical heating.
11. The apparatus of claim 1 , wherein the optical detector comprises a light source configured to emit light in an absorption band of a fluorescent dye into the amplification reaction zone when the multi-lane microfluidic cartridge is received in the receiving bay, and a light detector configured to detect light in an emission band of the fluorescent dye from the amplification reaction zone when the multi-lane microfluidic cartridge is received in the receiving bay, and wherein the fluorescent dye binds to a fluorescent polynucleotide probe or a fragment thereof.
12. The apparatus of claim 11 , wherein the optical detector is configured to selectively emit light in the absorption band of the fluorescent dye, and selectively detect light in the emission band of the fluorescent dye.
13. The apparatus of claim 1 , further comprising one or more components configured to inhibit motion of a material in the multi-lane microfluidic cartridge when the multi-lane microfluidic cartridge is received in the receiving bay.
14. The apparatus of claim 13 , wherein the one or more components configured to inhibit motion of the material comprises a valve configured to transition from an open state to a closed state when the multi-lane microfluidic cartridge is received in the receiving bay.
15. The apparatus of claim 14 , wherein the valve is configured to allow the material to pass along a channel from a position on one side of the valve to a position on the other side of the valve when in the open state, and wherein the valve is configured to prevent the material from passing along the channel from the position on one side of the valve to the position on the other side of the valve when in the closed state.
16. The apparatus of claim 15 , wherein the material comprises a polynucleotide-containing sample.
17. The apparatus of claim 16 , wherein the one or more components configured to inhibit motion of the polynucleotide-containing sample comprises a plurality of valves configured to isolate the amplification reaction zone during cyclical heating when the multi-lane microfluidic cartridge is received in the receiving bay.
18. The apparatus of claim 15 , wherein the material comprises a gas.
19. The apparatus of claim 15 , wherein the material comprises a reagent.
20. The apparatus of claim 1 , wherein the multi-lane microfluidic cartridge is disposable.
21. The apparatus of claim 1 , wherein the multi-lane microfluidic cartridge comprises a waste reservoir and a channel extending between the processing chamber and the waste reservoir.
22. The apparatus of claim 21 , further comprising a valve configured to inhibit motion of a material in the channel when the multi-lane microfluidic cartridge is received in the receiving bay.
23. The apparatus of claim 1 , further comprising:
a second receiving bay configured to receive a second multi-lane microfluidic cartridge;
wherein the apparatus is configured to concurrently process a plurality of samples in the first multi-lane microfluidic cartridge and the second multi-lane microfluidic cartridge when the first and second multi-lane cartridges are received in the receiving bay and the second receiving bay, respectively, and
wherein the apparatus is configured to consecutively process a plurality of samples in the first multi-lane microfluidic cartridge and the second multi-lane microfluidic cartridge when the first and second multi-lane cartridges are received in the receiving bay and the second receiving bay, respectively.