Channels with cross-sectional thermal gradients

• US 9,222,115 B2
• Filed: 12/21/2012
• Issued: 12/29/2015
• Est. Priority Date: 12/30/2011
• Status: Expired due to Fees

• Current Assignee: Abbott Molecular Incorporated (Abbott Laboratories)
• Original Assignee: Abbott Molecular Incorporated (Abbott Laboratories)
• Inventors: Marble, Herbert A., Laffler, Thomas
• Primary Examiner(s): Kim, Young J
• Application Number:
  US13/724,503
• Publication Number:
  US 20130183717A1
• Time in Patent Office: 1,103 Days
• US Class Current: 1/1
• CPC Class Codes: B01L 2400/0415 electrical forces, e.g. electrokinetic
• CPC Class Codes: B01L 2400/0433 vibrational forces
• CPC Class Codes: B01L 2400/0445 Natural or forced convection
• CPC Class Codes: B01L 2400/086 using baffles or other fixed flow obstructions
• CPC Class Codes: B01L 3/502784 specially adapted for droplet or plug flow, e.g. digital microfluidics automatic analysis using a stream of discrete samples in a tube system G01N35/08
• CPC Class Codes: B01L 7/525 with physical movement of samples between temperature zones
• CPC Class Codes: B01L 7/54 using spatial temperature gradients
• CPC Class Codes: C07H 21/00 Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
• CPC Class Codes: C12P 19/34 Polynucleotides, e.g. nucleic acids, oligoribonucleotides
• CPC Class Codes: C12Q 1/686 Polymerase chain reaction [PCR]
• CPC Class Codes: C12Q 2527/15 Gradients
• CPC Class Codes: C12Q 2563/159 Microreactors, e.g. emulsion PCR or sequencing, droplet PCR, microcapsules, i.e. non-liquid containers with a range of different permeability's for different reaction components
• CPC Class Codes: C12Q 2565/629 being a microfluidic device

Abstract

Provided herein are systems, devices, and methods for generating thermal gradients in channels and uses thereof. In particular, provided herein are system, methods, and devices employing first and second thermal layers positioned around a channel in order to create a thermal gradient across a cross-section of the channel having, for example, a nucleic acid denaturation zone, a nucleic acid annealing zone, and a nucleic acid polymerization zone. Such devices find use in, for example, nucleic acid amplification procedures, including digital polymerase chain reaction (dPCR) to temperature cycle droplets for amplification of nucleic acid templates within the droplets.

Claims

1. A device comprising:

a) a microchannel comprising a surface configured for transmission of a carrier medium comprising droplets;

b) a first thermal layer positioned along said microchannel configured to heat a portion of said microchannel surface to a nucleic acid denaturation temperature; and

c) a second thermal layer positioned along said microchannel configured to heat or cool a portion of said microchannel surface to a nucleic acid annealing temperature;

wherein said first and said second thermal layers are positioned along opposing surface walls of said microchannel to create a thermal gradient across a cross-section of said microchannel; and

wherein said thermal gradient comprises a nucleic acid denaturation zone, a nucleic acid annealing zone, and a nucleic acid polymerization zone.

2. The device of claim 1, further comprising a plurality of motion components positioned along said microchannel, wherein said plurality of motion components are configured to impart motion to said droplets in said carrier medium when present in said microchannel such that stochastic contact of said droplet with said microchannel surface is increased.

3. The device of claim 2, wherein said motion is imparted via mechanical agitation.

4. The device of claim 3, wherein said mechanical agitation is selected from the group consisting of:

vibration, low-frequency sonication, and acoustic waves.

5. The device of claim 2, wherein said motion is imparted via convection and/or turbulent flow.

6. The device of claim 2, wherein said motion is imparted via an electrical field.

7. The device of claim 2, wherein said droplets comprise paramagnetic microparticles, and wherein said plurality of motion components are configured to induce an oscillating magnetic field along said channel.

8. The device of claim 2, wherein said plurality of motion components are positioned intermittently along said microchannel.

9. The device of claim 1, further comprising said droplets in said carrier medium.

10. The device of claim 1, wherein said droplets each comprise at least one component selected from the group consisting of:

primers, a nucleic acid template, a polymerase, and dNTPs.

11. The device of claim 1, further comprising an optical array sensor positioned along said microchannel configured to monitor PCR signal output in said droplets.