Canon U.S. Life Sciences, Inc. (Rockville, MD)

This invention is about methods and devices that control an integrated thin film device that has a plurality microfluidic channels. One embodiment of the microfluidic devices includes microfluidic chips, which have a first zone with a plurality microfluidic channel and a second zone that has a number of microfluidic channels. The microfluidic channel within the second zone is connected to fluids. A thin-film heater is also part of the microfluidic device. It is communicating thermally with microfluidic channels of the first and the second zones. The microfluidic device is also equipped with the control system which can independently control the temperature of each of the heaters made of thin film using pulse width modulation (PWM) control signals that are optimized for each of the heaters made of thin-film.

1. The Field of Invention

The present invention relates to microfluidic devices and the temperature control of microfluidic devices used for biological reactions. Particularly, the present invention relates to systems and techniques for controlling and determining the temperature of integrated thin-film resistive heater elements within the microfluidic device.

2. Discussion of the Background

The detection of nucleic acids is central to medicine as well as forensic science, industrial processing as well as animal breeding, as well as many other fields. The ability to detect disease conditions (e.g., cancer), infectious organisms (e.g., HIV), geneticlineage, genetic markers, and the like, is ubiquitous technology for disease diagnosis and prognosis, marker assisted selection, identification of crime scene features, the ability to propagate industrial organisms and many other techniques.Determination of the integrity of a nucleic acid of interest can be relevant to the pathology of an infection or cancer.

An easy and effective method to detect tiny amounts of nucleic acid is by replicating a portion or all of a sequence of nucleic acid many times, and then studying the amplification products. Polymerase chain reaction (PCR) is awell-known technique to amplify DNA. One template DNA molecule can be used to generate millions of copies of DNA with the PCR. The PCR process includes stages of “denaturation,” “annealing,” and “extension.” These phases are part a cycle that is repeated many times until there are enough copies are available to be detected and analysed at conclusion. Sambrook and Russell, Molecular Cloning – A Laboratory Manual (3rd Edition) give more details on PCR. ), Vols. 3 and 4, Cold SpringHarbor Laboratory, Cold Spring Harbor, N.Y. (2000); Current Protocols in Molecular Biology, F. M. Ausubel et al. in the editor’s chair., Current Protocols, jointly owned by Greene Publishing Associates, Inc. and John Wiley & Sons, Inc., (supplemented through2005) and PCR Protocols A Guide to Methods and Applications, M. A. Innis and M. A. Innis. in the editor’s chair., Academic Press Inc. San Diego, Calif. (1990).

The PCR processes of denaturing, annealing and extension take place at various temperatures and cause targets DNA molecules to duplicate themselves. The temperature cycling (thermocyling), requirements for specific nucleic acids samples and assays will vary. In the denaturing phase, double-stranded DNA (dsDNA) is separated by heat into single stranded DNA (ssDNA). Primers are joined to single-strand DNA molecules during the annealing phase. Single-strand DNA molecules are grown to double strand DNA in the extension phase through specific bindings between nucleotides in the PCR solution and single DNA strand. Typical temperatures are C. for denaturing, C. to anneal, C. forextension. The temperature is maintained during each phase for a specific amount of time which may be a fraction of one second to 10 seconds or less. The DNA is amplified at every cycle. It typically requires between 20 and 40 cycles to produce enough DNA for theapplications. It is crucial to manage accurately the temperature of the samples during different phases in order to achieve an adequate yield.

In recent times, a variety of high-throughput techniques for accomplishing PCR as well as other amplification reactions have been developed, e.g., involving Amplification reactions that are performed in microfluidic devices and methods for the detection and analysis of amplified nucleic acid within or in the devices. Thermal cycling of the sample for amplification can be usually accomplished in two ways. In the first method the solution of the sample is placed in the instrument, and then the temperature regulated in time, muchlike the conventional PCR device. Another method is when the sample solution is continuously moved through temperature zones that vary spatially. Lagally and colleagues, for example. (Analytical Chemistry 73:565-570 (2001)), Kopp et al. (Science280:1046-1048 (1998)), Park et al. (Analytical Chemistry 75:6029-6033 (2003)), Hahn and others. (WO 2005/075683), Enzelberger et al. (U.S. Pat. No. No. 6.960,437) and Knapp (et and. (U.S. Patent Application Publication No. 2005/0042639).

A variety of detection techniques require a specific amount of copies (millions, for example) of the DNA molecule, in order for the DNA to be studied. The time required for the process is only decreased if the length of the cycle is shorter. So, the total process time can be drastically decreased by quickly warming and cooling the samples to achieve the desired temperature while ensuring they are maintained for the duration of the phase.

Accordingly, what is desired is a method to rapidly and precisely alter the process temperature in PCR.

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