Micro-nano system fluid chip detection system and detection method

Abstract

The invention relates to a detection system and detection method of a micro-nanoliter system fluid chip, which is characterized in that it includes an incident light generation system, a bifocal plane fluorescence collection system, a collection light processing system, a circulating flow three-dimensional temperature-controlled stage, and a micro Fluid chip; the incident light generation system at least includes a light source and a condenser lens arranged at the output end of the light source; the bifocal fluorescence collection system includes two imaging lens groups, front and rear, and a detector arranged at the output end of the rear lens group; light collection processing The system includes an A / D acquisition card connected to the output end of the detector, and a computer connected to the output end of the A / D acquisition card; the temperature-controlled stage includes a movement platform carrying a microfluidic chip, and surrounding movement platforms The heat transfer medium, the heater arranged around the heat transfer medium and the heat preservation device arranged outside the heater; the computer connects the motion platform through a motion controller, and connects the heater and the temperature sensor through a temperature control feedback circuit. The invention has the advantages of simple structure, low cost, high fluorescence collection efficiency and detection sensitivity, and convenient operation and use, making the invention more favorable for promotion and popularization.

Description

technical field [0001] The invention relates to a detection method and system of a microfluidic chip, in particular to a detection method and system of a micro-nanoliter system (1nL-10μL) fluid chip with low cost and relatively high detection sensitivity and resolution. Background technique [0002] Microfluidic chip technology, also known as "Lab-on-a-chip", or micro-separation in Miniaturized Total Analysis System (μTAS: Micro-separation in Miniaturized Total Analysis System), is an emerging technology in the late 20th century. It is a multidisciplinary interdisciplinary technology, which uses system integration engineering technologies such as micro-electromechanical processing, computers, electronics, materials science, optics, mechanics, automatic control, software, etc. and multidisciplinary application fields such as analytical chemistry, biology, and medicine to carry out organic research. Cross-combination integrates discontinuous processes such as sample preparatio...

AI Technical Summary

Problems solved by technology

Fluorescence microscope is a hand-operated and easy-to-use microfluidic chip detection device, which can observe single-channel low-resolution microfluidic chip reaction signals, and the observed fluorescent signals can be converted to The disadvantages of collecting and storing digital images are that the observation range is relatively small, generally within a few millimeters, the detection speed is relatively slow (<100Hz), and the cost of the refrigerated CCD digital fluorescence microscope is relatively expensive.

Confocal scanner is an internationally popular automated microfluidic chip detection instrument, which can perform multi-channel, large area (100 mm × 100 mm), medium resolution (5 μm to 50 μm), relatively fast detection speed ( <100kHz) microfluidic chip signal detection and storage to generate digital images, but the structure of the confocal scanner is complex and the cost is relatively expensive

Electrochemical detection is an important microfluidic chip detection method. According to the principle and application characteristics of electrochemical detection, the electrochemical detector is made together with the microchip, and the microfluidic chip is integrated with the amperometric method, conductometric method, and potentiometric method. The detection signal is amplified and digitally output, and the electrochemical detection is easy to realize the miniaturization of the microfluidic chip detection system, but the detection sensitivity is not very high

Real-time fluorescent quantitative PCR (polymerase chain reaction, polymerase chain reaction) instrument is also an important microfluidic detection instrument, but it is still limited to use Tube tubes for microfluidic reaction detection of systems above 5 μL, and the instrument price is relatively expensive. Need more than 300,000 yuan / set

[0004] Many domestic units have carried out a lot of work on test tube-based real-time fluorescence quantitative PCR detection, such as "application number 200410073432.1", "application number 200510028623.0", "application number 01139052.2", "patent number ZL200420102062.5", etc. Their core research The work has one thing in common, which is to use different optical fiber (single, multiple, or array) coupling methods to detect real-time fluorescence quantitative PCR amplification reaction signals in test tubes, so as to simplify the structure of the optical path system and reduce the cost of the entire instrument. Purpose, due to the coupling and signal transmission of the optical fiber, the numerical aperture of the receiving end of the fluorescence signal cannot be very high, and the loss of the fluorescence signal increases significantly with the increase of the distance when the fluorescence signal is transmitted in the optical fiber, so the detection sensitivity of the instrument is greatly limited , which is not as high as the detection sensitivity obtained by the conventional lens combination method, which provides opportunities and development space for the innovative research work on the microfluidic chip detection technology using the lens combination method

Some people ("Application No. 01128122.7") conducted research on the integration of nucleic acid amplification and gene chip hybridization. No specific innovation work content proposed

Also, someone introduced a temperature-changing method for setting a plurality of different temperature regions in the microfluidic chip application packaging structure for PCR amplification ("Patent No. ZL200420072774.7") to allow samples to pass through these regions back and forth to achieve PCR amplification. Amplifier (“Application No. 01120216.5”) introduced a complex heating temperature control device, the purpose of which is to ensure the uniformity of the heat source, and to achieve uniform temperature distribution inside the PCR amplification cavity, but these work There are certain limitations, and there is no research content related to the amplification signal detection technology, and the system integration of temperature control and detection cannot be realized, and the use requirements of the gene amplification reaction detection system on the chip cannot be met.

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