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US 2014/0370492 A1Published application
Liquid reflux high-speed gene amplification device
AssigneesKanagawa Academy of Science and Technology, On-Chip Cellomics Consortium, National University Corporation Tokyo Medical and Dental University
InventorsKenji Yasuda, Hideyuki Terazono, Akihiro Hattori
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Abstract
The present invention provides a liquid reflux reaction control device including an additional mechanism that allows more stable temperature control, a pre-treatment mechanism that performs pre-treatment including a pre-PCR reaction reverse transcription reaction process that allows RNA detection, a melting curve analysis function, chip technology optimal for holding liquid droplets and optical measurement and the optical measurement function for PCR, and a temperature gradient control mechanism using a quantitative infrared light irradiation/absorption control technique.
Claims42
A liquid reflux reaction control device, comprising:
- a reaction vessel including one or a plurality of wells for containing a sample liquid;
- a reaction vessel casing that covers the reaction vessel in a sealing manner so as to prevent droplets of the sample liquid located in the well(s) from evaporating and includes a heat-retainer for preventing dew condensation;
- a heat exchange vessel that is provided in contact with the reaction vessel so as to conduct heat to the reaction vessel and includes an inlet and an outlet respectively for introducing and discharging a liquid of a predetermined temperature;
- a plurality of liquid reservoir tanks each provided with a temperature-controllable heat source for maintaining the liquid contained therein at a predetermined temperature, a liquid stirring mechanism that stirs the liquid in the reservoir tank so as to uniformize the temperature of the liquid, and a temperature sensor for providing feedback information for controlling the temperature of the liquid in the reservoir tank;
- a thin tube that connects the plurality of liquid reservoir tanks to each other in a fluid-communicable manner to adjust liquid surface levels of the plurality of liquid reservoir tanks to be substantially the same;
- a tubular flow channel that connects the inlet or the outlet of the heat exchange vessel to each of the liquid reservoir tanks;
- a pump that is provided on the tubular flow channel and is capable of circulating the liquid at a rate 10 mL/sec. or higher between the heat exchange vessel and each of the liquid reservoir tanks;
- a switching valve that is provided on the tubular flow channel and controls a flow of the circulating liquid, the switching valve switching a flow of the liquid of the predetermined temperature from each of the plurality of liquid reservoir tanks into the heat exchange vessel at a predetermined time interval to control the temperature of the reaction vessel to a desired temperature;
- an auxiliary temperature control mechanism that is located on the tubular flow channel between the heat exchange vessel and the liquid reservoir tanks, has a predetermined capacity that allows the liquid that is refluxing to be temporarily held therein, and refluxes the liquid to the liquid reservoir tank after adjusting the temperature of the liquid to the temperature of the liquid reservoir tank so as to minimize temperature change in the liquid reservoir tank;
- a fluorescence detector that, in the case where the sample liquid contains a fluorescent dye, detects fluorescence emitted by the fluorescent dye in the well(s) in association with an operation of the switching valve of switching the temperature of the reaction vessel so as to measure time-wise change in the intensity of the fluorescence;
- and
- a control analyzer capable of estimating the temperature of the sample liquid based on the fluorescence intensity and controlling an operation of the switching valve based on the estimation result;
- wherein the sample has an amount of several ten microliters per well or smaller, and the liquid to be circulated has a total volume of several ten milliliters per liquid reservoir tank or larger.
10 more elements
A liquid reflux reaction control device, comprising:
- a sample holder including one or a plurality of wells for holding a sample liquid;
- a laser device that emits infrared laser light which is absorbable to water as the sample liquid;
- a gray-scale ND filter discus capable of continuously changing the intensity of the laser light from the laser device;
- a rotation control mechanism that controls the rotation rate of the discus;
- an optical system for leading the laser light to the sample liquid in the well(s) via the gray-scale ND filter discus;
- a temperature control mechanism that controls the temperature of the well(s);
- and
- an optical measurement device including an optical camera that measures an optical image of the sample liquid in the well(s).
5 more elements
A liquid reflux reaction control device, comprising:
- a reaction vessel including one or a plurality of wells for containing a sample liquid;
- a heat exchange vessel that is provided in contact with the reaction vessel so as to conduct heat to the reaction vessel and includes an inlet and an outlet respectively for introducing and discharging a liquid of a predetermined temperature;
- a liquid reservoir tank provided with a temperature-controllable heat source and a temperature sensor for maintaining the liquid contained therein at a predetermined temperature;
- a tubular flow channel that connects the inlet or the outlet of the heat exchange vessel to the liquid reservoir tank;
- a pump, provided on the tubular flow channel, for circulating the liquid between the heat exchange vessel and the liquid reservoir tank;
- a laser device that emits infrared laser light which is absorbable to water as the sample liquid;
- a gray-scale ND filter discus capable of continuously changing the intensity of the laser light from the laser device;
- a rotation control mechanism that controls the rotation rate of the discus;
- an optical system for leading the laser light to the sample liquid in the well(s) via the gray-scale ND filter discus;
- and
- an optical measurement device including an optical camera that measures an optical image of the sample liquid in the well(s).
8 more elements
Dependencies are inferred from the claim text.