# US 10,871,460 B2 · Method and apparatus for generating thermal melting curves in a microfluidic device

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- Number: US 10,871,460 B2
- Document ID: US10871460B2
- Status: Granted
- Kind: B2
- Application number: 15/990,369
- Filed: 2018-05-25
- Published: 2018-11-08
- Granted: 2020-12-22
- Earliest claimed: 2000-11-16
- Assignees: Canon U.S.A., Inc.
- Applicants: Canon U.S.A., Inc.
- Inventors: Ivor T. Knight; Deborah John Boles; Aaron Rulison; Wesley B. Dong; Andrew Fabans; Allen Boronkay; Edward Donlon; Robert Moti; Michael Slater; Steven A. Sundberg; Michael R. Knapp
- Web page: https://patentgrep.com/patent/US10871460B2

## CPC

- G01N 25/04: of melting point; of freezing point; of softening point
- B01L 3/5027: {by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip}
- B01L 3/502715: {characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces}
- B01L 3/50273: {characterised by the means or forces applied to move the fluids}
- B01L 7/52: {with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples}
- B01L 7/525: {with physical movement of samples between temperature zones}
- C12Q 1/68: involving nucleic acids
- C12Q 1/6844: Nucleic acid amplification reactions
- C12Q 1/686: Polymerase chain reaction [PCR]
- C12Q 1/6862: Ligase chain reaction [LCR]
- G01N 21/33: using ultraviolet light (G01N21/39 takes precedence)
- G01N 21/6428: {Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" (in vivo A61B5/00; immunoassay G01N33/53)}
- B01L 2200/10: Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
- B01L 2300/0627: Sensor or part of a sensor is integrated
- B01L 2300/0816: Cards, e.g. flat sample carriers usually with flow in two horizontal directions
- B01L 2300/14: Means for pressure control
- B01L 2300/18: Means for temperature control
- B01L 2300/1833: using electrical currents in the sample itself
- B01L 2400/0409: centrifugal forces
- B01L 2400/0415: electrical forces, e.g. electrokinetic
- B01L 2400/0487: fluid pressure, pneumatics
- Y10T 436/143333: Saccharide [e.g., DNA, etc.]
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)
- B01L: CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- Y10T: TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION

## Abstract

The present invention provides novel methods and devices that employ microfluidic technology to generate molecular melt curves. In particular, the devices and methods in accordance with the invention are useful in providing for the analysis of PCR amplification products.

## Claims

### Claim 1

A method of determining the thermal inertia of a microfluidic device, the method comprising:
flowing a fluid comprising a fluorescent material through a channel in a microfluidic device, wherein the amount of fluorescence generated by the fluorescent material varies with temperature,
varying the temperature of an external surface of the microfluidic device, wherein the variation comprises a pattern with a recognizable feature, wherein the variation in temperature of the external surface produces a corresponding variation in temperature within the channel, and wherein the variation in temperature in the channel produces a corresponding variation in the amount of fluorescence generated by the fluorescent material,
measuring the fluorescence generated by the fluorescent material in the channel, and
determining a time offset between the temperature variations of the external surface and the variations in the amount of fluorescence by measuring the time delay between the imposition of the recognizable feature on the external surface and the appearance of the recognizable feature in the fluorescence generated by the fluorescent material, whereby the time offset is indicative of the thermal inertia.

### Claim 2 (depends on 1)

The method of claim 1, wherein the pattern is a sine wave.