Fluorescence detector for microfluidic diagnostic system
The present technology provides for a fluorescent detector that is configured to detect light emitted for a probe characteristic of a polynucleotide. The polynucleotide is undergoing amplification in a microfluidic channel with which the detector is in optical communication. The detector is configured to detect minute quantities of polynucleotide, such as would be contained in a microfluidic volume. The detector can also be multiplexed to permit multiple concurrent measurements on multiple polynucleotides concurrently.
1 . A method comprising:
positioning a consumable microfluidic cartridge above a plurality of amplification heaters, wherein the plurality of amplification heaters are arranged along an amplification heater axis, the plurality of amplification heaters comprising at least a first amplification heater and a second amplification heater, the consumable microfluidic cartridge comprising a plurality of detection regions, the plurality of detection regions comprising at least a first detection region and a second detection region, the plurality of detection regions arranged along a first cartridge axis, wherein the amplification heater axis and the first cartridge axis are substantially parallel when the consumable microfluidic cartridge is positioned above the plurality of heaters;
amplifying one or more polynucleotides in the first detection region by directing heat toward the first detection region with the first amplification heater when the consumable microfluidic cartridge is positioned above the plurality of amplification heaters, and amplifying one or more polynucleotides in the second detection region by directing heat toward the second detection region with the second amplification heater when the consumable microfluidic cartridge is positioned above the plurality of amplification heaters;
positioning a fluorescence detector above the consumable microfluidic cartridge, the fluorescence detector comprising a plurality of detectors arranged along a first detection axis, the plurality of detectors comprising a first detector and a second detector, wherein the first cartridge axis and the first detection axis are substantially parallel when the fluorescence detector is positioned above the consumable microfluidic cartridge; and
detecting the presence of one or more polynucleotides in the first detection region of the consumable microfluidic cartridge by transmitting light onto the first detection region and receiving light emitted from the first detection region by the first detector of the fluorescence detector, and detecting the presence of one or more polynucleotides in the second detection region of the consumable microfluidic cartridge by transmitting light onto the second detection region and receiving light emitted from the second detection region by the second detector of the fluorescence detector, and
disposing the consumable microfluidic cartridge and reusing the plurality of amplification heaters and the fluorescence detector by positioning a second consumable microfluidic cartridge above the plurality of amplification heaters and positioning the fluorescence detector above the second consumable microfluidic cartridge.
2 . The method of claim 1 , further comprising moving the fluorescence detector relative to the plurality of detection regions of the consumable microfluidic cartridge.
3 . The method of claim 1 , further comprising moving the fluorescence detector in a direction parallel to the amplification heater axis and the first cartridge axis.
4 . The method of claim 1 , further comprising moving the fluorescence detector along the first detection axis.
5 . The method of claim 1 , further comprising transmitting light onto the first detection region with a light source of the first detector, and receiving light emitted from the first detection region by a photodiode of the first detector.
6 . The method of claim 1 , further comprising transmitting light onto the first detection region with a light source of the first detector that selectively emits light in an absorption band of a fluorescent dye, and receiving light emitted from the first detection region by a light detector of the first detector that selectively detects light in an emission band of the fluorescent dye.
7 . The method of claim 6 , wherein the fluorescent dye corresponds to a fluorescent polynucleotide probe or a fragment thereof.
8 . The method of claim 1 , further comprising transmitting light onto the first detection region with a bandpass-filtered diode of the first detector, and receiving light emitted from the first detection region by a bandpass-filtered photodiode of the first detector.
9 . The method of claim 1 , further comprising transmitting a diagnostic result to a user.
10 . The method of claim 1 , wherein the plurality of detection regions comprise twelve detection regions, further comprising amplifying one or more polynucleotides in twelve detection regions.
11 . The method of claim 1 , wherein the plurality of detection regions comprise twenty-four detection regions, further comprising amplifying one or more polynucleotides in twenty-four detection regions.
12 . The method of claim 1 , further comprising simultaneously detecting the presence of one or more polynucleotides in the first detection region and the second detection region.
13 . The method of claim 1 , further comprising loading a first inlet with a first sample, and loading a second inlet with a second sample that is different than the first sample, wherein the first inlet is in fluid communication with the first detection region and the second inlet is in fluid communication with the second detection region.
14 . The method of claim 1 , wherein amplifying one or more polynucleotides in the first detection region further comprises heating through a series of cycles with the first amplification heater.
15 . The method of claim 1 , wherein amplifying one or more polynucleotides in the first detection region further comprises subjecting a sample to a temperature protocol that causes one or more reactions to occur.
16 . The method of claim 2 , wherein amplifying one or more polynucleotides in the first detection region further comprises maintaining a desired speed of heating.
17 . The method of claim 1 , further comprising positioning an aperture plate above the consumable microfluidic cartridge.
18 . The method of claim 1 , further comprising applying pressure to the consumable microfluidic cartridge.
19 . The method of claim 1 , wherein amplifying one or more polynucleotides in the first detection region by directing heat toward the first detection region with the first amplification heater comprises heating with a resistive heater.
20 . The method of claim 1 , further comprising scanning a barcode associated with a sample.