Integrated system for processing microfluidic samples, and method of using 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:
an automated sampling device;
a reagent reservoir;
a platform configured to support a multi-lane microfluidic cartridge configured to process the plurality of polynucleotide-containing samples, 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 a region where amplification occurs, 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 a first sample of the plurality of polynucleotide-containing samples is passed within the multi-lane microfluidic cartridge at a different time than a second sample of the plurality of polynucleotide containing samples;
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 supported by the platform, 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;
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 supported by the platform, 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 amplification is performed in an automated manner;
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 supported by the platform, 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 within a region where amplification occurs at each temperature of the at least two temperatures when the multi-lane microfluidic cartridge is supported by the platform;
wherein the plurality of second heat sources are configured to perform amplification reactions in the plurality of sample lanes when the multi-lane microfluidic cartridge is supported by the platform;
wherein a sample lane of the multi-lane microfluidic cartridge comprises a processing chamber, and wherein the first heat source is configured to apply heat to the processing chamber when the multi-lane microfluidic cartridge is supported by the platform;
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 when the multi-lane microfluidic cartridge is supported by the platform and the polynucleotide-loaded retention member is received in the multi-lane microfluidic cartridge; and
an optical detector configured to detect the presence of the one or more amplified polynucleotides within the region where amplification occurs.
2. The apparatus of claim 1 , wherein the reagent reservoir comprises a wash buffer reservoir, and wherein the plurality of sample lanes are configured to share a wash buffer received in the multi-lane microfluidic from the wash buffer reservoir.
3. The apparatus of claim 1 , wherein the reagent reservoir comprises a release reagent reservoir, and wherein the plurality of sample lanes are configured to share a release reagent received in the multi-lane microfluidic cartridge from the release reagent reservoir.
4. The apparatus of claim 1 , wherein the reagent reservoir comprises a hydroxide solution reservoir, and wherein the plurality of sample lanes are configured to share a hydroxide solution received in the multi-lane microfluidic from the reagent reservoir.
5. The apparatus of claim 1 , further comprising the multi-lane microfluidic cartridge supported by the platform 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 supported by the platform.
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 supported by the platform 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 is fixed in position relative to the multi-lane microfluidic cartridge during cyclical heating.
9. 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 region where amplification occurs when the multi-lane microfluidic cartridge is supported by the platform, and a light detector configured to detect light in an emission band of the fluorescent dye from the region where amplification occurs when the multi-lane microfluidic cartridge is supported by the platform, wherein the fluorescent dye binds to a fluorescent polynucleotide probe or a fragment thereof.
10. The apparatus of claim 9 , 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.
11. 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 supported by the platform.
12. The apparatus of claim 11 , wherein the material comprises a polynucleotide-containing sample.
13. The apparatus of claim 12 , wherein the one or more components configured to inhibit motion of the polynucleotide-containing sample comprises a plurality of valves configured to prevent fluid flow from the region where amplification occurs during cyclical heating when the multi-lane microfluidic cartridge is supported by the platform.
14. The apparatus of claim 11 , wherein the material comprises a gas.
15. The apparatus of claim 11 , wherein the material comprises a reagent.
16. The apparatus of claim 1 , wherein the multi-lane microfluidic cartridge is disposable.
17. 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.
18. The apparatus of claim 17 , further comprising a valve configured to inhibit motion of a material in the channel when the multi-lane microfluidic cartridge is supported by the platform.
19. The apparatus of claim 1 , further comprising:
a second platform configured to support 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 supported by the platform and the second platform, respectively, and
wherein the apparatus is configured to successively 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 supported by the platform and the second platform, respectively.