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Publication Number
US-12030050-B2
Publication Date
2024-07-09
Expiration Date
Abstract
The present technology provides for a microfluidic substrate configured to carry out PCR on a number of polynucleotide-containing samples in parallel. The substrate can be a single-layer substrate in a microfluidic cartridge. Also provided are a method of making a microfluidic cartridge comprising such a substrate. Still further disclosed are a microfluidic valve suitable for use in isolating a PCR chamber in a microfluidic substrate, and a method of making such a valve.
Core Innovation
The invention relates to a microfluidic cartridge for isolating a plurality of polynucleotide-containing samples and performing amplification with improved thermal contact. The cartridge includes multiple reaction chambers and microfluidic valve sets associated with sample lanes, with inlets and channels leading to and from the reaction chambers. Different polynucleotide-containing samples are isolated within respective reaction chambers by closing the corresponding microfluidic valves.
To assist amplification, the method applies a minimum contact pressure of at least 1 psi to contact the microfluidic cartridge to a heater of an instrument, or applies a contact pressure of at least 1 psi by a contact force member to press the cartridge over heaters. This is used to assist achieving better thermal contact between the instrument heater and heat-receivable parts of the cartridge, and to thermally couple at least one heat source to one or more distinct locations on the cartridge.
The invention further includes downstream detection of amplified polynucleotides using an optical detector configured with a light source emitting in a fluorescent dye absorption band and a light detector detecting in a fluorescent dye emission band. The fluorescent dye corresponds to a fluorescent polynucleotide probe or a fragment thereof, while independent amplification may be performed in the first reaction chamber and the second reaction chamber while maintaining the isolation of each sample by independently closing the relevant valves.
Claims Coverage
The independent claims identified are 1, 10, and 16. Across these claims, the main inventive features total 3 clusters: isolating different polynucleotide-containing samples in separate microfluidic reaction chambers using closed microfluidic valves, applying at least 1 psi contact pressure to thermally couple heater(s) to distinct locations or heat-receivable parts for amplification, and detecting amplified nucleotides via fluorescence using fluorescent dye absorption/emission bands and a fluorescent polynucleotide probe or fragment thereof.
Isolation of different samples in separate reaction chambers by closing valve sets
Introducing a first polynucleotide-containing sample into a first reaction chamber via a first inlet; introducing a second polynucleotide-containing sample into a second reaction chamber via a second inlet, wherein the first sample is different than the second sample; isolating the first sample within the first reaction chamber with a first set of microfluidic valves closed; independently isolating the second sample within the second reaction chamber with a second set of valves closed.
Applying minimum contact pressure of at least 1 psi to improve thermal contact with heaters
Applying a minimum contact pressure of at least 1 psi over the microfluidic cartridge to contact the microfluidic cartridge to a heater of an instrument to assist in achieving better thermal contact between the heater of the instrument and heat-receivable parts of the microfluidic cartridge.
Microfluidic lane networks with first and second valves to prevent movement during isolation
Providing a microfluidic cartridge comprising a plurality of sample lanes, wherein each sample lane comprises a microfluidic network with a first valve, a second valve, a reaction chamber, a first channel leading via the first valve to the reaction chamber, and a second channel leading via the second valve from the reaction chamber; introducing first and second different polynucleotide-containing samples into respective reaction chambers in respective microfluidic networks; isolating each sample to prevent movement into or out of the corresponding reaction chamber by closing the first valve and the second valve of the respective microfluidic network.
Contact pressure of at least 1 psi to thermally couple heat source(s) to distinct locations
Applying a contact pressure of at least 1 psi to the microfluidic cartridge to thermally couple at least one heat source to one or more distinct locations on the microfluidic cartridge.
Pressing the cartridge over heaters with at least 1 psi contact force for thermal contact during amplification
Applying contact force of at least 1 psi by a contact force member to press the microfluidic cartridge over heaters so that the first reaction chamber and the second reaction chamber make thermal contact with the heaters for amplification.
Fluorescent detection using dye absorption and emission bands with a fluorescent polynucleotide probe
Detecting amplified nucleotides using an optical detector with a light source emitting in a fluorescent dye absorption band and a light detector detecting in the fluorescent dye emission band, wherein the fluorescent dye is a fluorescent polynucleotide probe or fragment thereof.
The independent claims collectively cover isolating different polynucleotide-containing samples in separate microfluidic reaction chambers using closed microfluidic valves, while applying a contact pressure of at least 1 psi to thermally couple heater(s) to the cartridge. Fluorescent optical detection with dye absorption/emission bands using a fluorescent polynucleotide probe or fragment thereof is claimed as a refinement in the claim set.
Stated Advantages
Assist in achieving better thermal contact between the heater of the instrument and heat-receivable parts of the microfluidic cartridge.
Documented Applications
No documented applications found
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