CN103257213A - Totally-integrated high-flux cell level micro-fluidic chip medicine evaluating system - Google Patents
Totally-integrated high-flux cell level micro-fluidic chip medicine evaluating system Download PDFInfo
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Abstract
A totally-integrated high-flux cell level micro-fluidic chip medicine evaluating system is a multifunctional cell level medicine evaluating system treating a micro-fluidic chip as a core technology, starting from the total design of an integrated high-flux automatic micro-fluidic chip system and integrating a cell function chip module, a flow path control module, a cell culture microenvironment module and a cell response signal real-time monitoring module. The totally-integrated high-flux cell level micro-fluidic chip medicine evaluating system has the characteristics of total integration, high flux, less sample consumption, totally-closed operation, realization of multi-parameter real time monitoring, and the like, has a very strong original innovativeness, and is of great significance to the large-scale medicine research and development.
Description
Technical field
The invention belongs to cellular level drug evaluation technical field, the horizontal micro-fluidic chip drug evaluation of a kind of fully integrated high-flux cell system is provided especially.
Background technology
Evaluating drug effect and Study on mechanism are the important steps of medicine innovation, namely determine its therapeutic action and general pharmacological action by observation and research to the compound candidate mechanism of action, for the new drug clinical testing provides reliable basis.The conventional medicament evaluation method is because of animal and animal, the limitation of the attribute difference between the animals and human beings body and assessment technique means and cause poor efficiency in the new drug development and expensive, and this has become one of bottleneck of medicament research and development.Present existing multiple drug evaluation model, because the cellular level evaluation is more close to physiological condition, the accuracy rate height, disturbing factor is few, is the main models of carrying out drug evaluation.At present, the major technique approach that the cellular level drug evaluation adopts is the array microwell plate, carry out cell at microwell plate and cultivate, apply medicine irritation, and be aided with automation equipment and high resolving power fluorescence digital image system experimentize operation and data acquisition, analysis.Cellular level drug evaluation based on this form remains in many technical matterss, as the orifice plate fixed size, only can under static schema, operate, can't realize the automatic control of convection cell, simultaneously supporting automatic operating equipment and detection system pattern is single with it, can't carry out flexible transformation at various cell function features of experiment, be difficult to more realize that cell is to the real-time monitoring of outside stimuli responsive process.Therefore, can only obtain the end results of cellular response information in principle, and be difficult to obtain cell to real-time dynamic response information and other biological information of drug response.What is more important, used apparatus mostly depends on import at present, although its automaticity makes moderate progress, reagent consumption is big, equipment volume is huge, and expensive, has formed external businessman and enterprise to the monopolization situation of this market segment.Therefore, constantly occur at new drug target, new drug kinds such as biotech drug constantly increase, the competition of new drug development field is growing more intense under the new situation, set up and development has independent intellectual property right, has integrated, high flux, detects in real time, the horizontal drug evaluation systems face of novel cell of robotization feature exigence very, this also is field problem demanding prompt solutions such as China's new drug development and pharmacy evaluation.
Microfluidic system, refer to integrated or be integrated into substantially on the chip of more than square centimeters (even littler) basic operation units such as specimen preparation related in the fields such as biological and chemical, reaction, separation, detections, form network by the microchannel, run through total system with controlled fluid, in order to a kind of technology of the various functions that replace conventional biological or chemical laboratory.Its essential characteristic and sharpest edges are that flexible combination and the scale of multiple monotechnics on small platform is integrated.The peculiar micro-dimension feature of micro-fluidic chip and cell size are complementary, and the integrated feature of chip design flexible and scale is fit to carry out cell function research very much in addition.Particularly, because multidimensional network feature and the integrated feature of function of micro-fluidic chip might make conventional evaluating drug effect and Study on mechanism process integration finish at a very little chip.The micro-fluidic chip research development in recent years that with the cell is object is very fast, showing as on the one hand on the chip various and cell controls relevant monotechnics such as Micropump, little valve, light tweezer, electrophoresis etc. and reaches its maturity, then be embodied in the core technology relevant with cell research such as cell cultivation, cell sorting, lysis on the other hand and grasp substantially, conventional cell is cultivated, process integration such as cell is excited, cell marking and detection finish at a very little chip.The scale of various monotechnicss is integrated to make dozens or even hundreds of group to test parallel carrying out, and learns information to obtain large number of biological.
So far, international pharmaceutical industry has been recognized the great potential that microflow control technique contains, has the people in the industry to foretell that microflow control technique can occupy critical role undoubtedly in future drugs discovery field.Existing researcher attempts micro-fluidic chip is combined with patch clamp technique, and being used for the ion channel is the drug research of action target spot.Also there is the researcher to concentrate on the research of chip stream controlling party face, and attempts solving actual biological question.But generally speaking, micro-fluidic chip system is used for evaluating drug effect and Study on mechanism still is in the starting stage, and the work of the aspects such as selection, design, optimization and practical application of chip platform is also really launched, and detected object is scattered relatively and single.Particularly comprise multi-functional integrated micro-flow control drug evaluation systematic studyes such as fluid control module, cell cultivation microenvironment module for development and still be not reported, also not seeing has related application.
Based on above-mentioned present situation, the inventor is core technology with the micro-fluidic chip, making up the horizontal drug evaluation of a fully integrated automatic cytological system that integrates cell function chip module, stream control module, cell cultivation microenvironment module and cellular response real time monitoring signals module has important practical significance, this solves with the closely-related critical bottleneck problem in field, medical forward position significant for towards great actual demand.Compared with prior art, this system can not only provide more near microenvironment in the body of physiological condition for cell growth, and can different operating unit flexible combination, scale is integrated, realizes full automatic working down computer programming auxiliary, shorten evaluation time, reduce the actual consumption of cell.In addition, by changing the functionalization micro-flow control chip module, this system also can realize multiparameter, high-throughput drug evaluation, and to the real-time tracing analysis of cellular response signal, the various real-time cellular response information that its expection may be collected be prior art and instrument can't obtain.
Summary of the invention
The purpose of this invention is to provide the horizontal micro-fluidic chip drug evaluation of a kind of fully integrated high-flux cell system, this system has fully integrated, the flux height, and the sample consumption is few, and totally-enclosed operation can be carried out characteristics such as multiparameter monitoring in real time.
The invention provides the horizontal micro-fluidic chip drug evaluation of a kind of fully integrated high-flux cell system, this system is made up of with fluid control module, cell cultivation microenvironment control module, four modules of the real-time monitoring modular of cell fluorescence response signal with function chip module, drug evaluation drug evaluation.
The horizontal micro-fluidic chip drug evaluation of fully integrated high-flux cell provided by the invention system, described drug evaluation is a kind of micro-fluidic chip based on PDMS polymeric material and soft etching technique with the function chip module, formed by 1～10 (preferred 4) identical structural unit, each structural unit can detect the effect of 1～10 kind of (preferred 3 kinds) medicine, by accurate pump valve control, can form the combination of multiple drug concentration gradient or different pharmaceutical.
The horizontal micro-fluidic chip drug evaluation of fully integrated high-flux cell provided by the invention system, described drug evaluation fluid control module is the fluid control systems of micro-fluidic chip, formed by the little valve/Micropump on array micro syringe pump and the chip, in the auxiliary Automatic Control that realizes down of computer programming, its function for the interpolation that realizes liquid such as nutrient culture media, medicine on the chip, transport and stream control such as concentration adjustment.
The horizontal micro-fluidic chip drug evaluation of fully integrated high-flux cell provided by the invention system, described cell is cultivated the microenvironment control module and is made of the cell culture chamber that can be fixed in microscope stage, gas control system, temperature humidity control system, chip places in this culturing room, keeps cell cultivation temperature, humidity and CO stable in it by a series of sensors and control module
2Gas concentration is achieved the monitoring in real time of the online cultivation of continuous cell and cell signal.
The horizontal micro-fluidic chip drug evaluation of fully integrated high-flux cell provided by the invention system, the real-time monitoring modular of described cell fluorescence response signal adopts the confocal fluorescent detection technique of automatically controlled scanning to realize that this module mainly comprises automatically controlled fluorescence imaging platform, ccd signal acquisition system, data handling system three parts.
The present invention has following advantage:
1. can realize the real-time tracing analysis to the cellular response signal.
2. cell and reagent consumption are few, can realize multiparameter, high-throughput drug evaluation.
3. can realize full-automatic, totally-enclosed operation down the auxiliary of computer programming, shorten evaluation time, reduce the cell contamination probability.
Description of drawings
The composition of the fully integrated high flux micro-fluidic chip of Fig. 1 drug evaluation system;
Fig. 2 drug evaluation function chip design synoptic diagram, wherein, a chip synoptic diagram, b partial enlarged drawing;
Fig. 3 cell is cultivated the microenvironment module diagram, and wherein, a module diagram, b cell are cultivated micro-objective table synoptic diagram;
The fully integrated high flux micro-fluidic chip of Fig. 4 drug evaluation system schematic.
Embodiment
Following examples will give further instruction to the present invention, but not thereby limiting the invention.
Embodiment
According to the fully integrated high flux micro-fluidic chip drug evaluation system of building shown in Figure 1, this system is made up of four modules.At first, according to making drug evaluation functional chip shown in Figure 2, chip is separatory road floor, PDMS thin layer, gas circuit floor and glass from top to bottom.Three first layers is the PDMS material, and using plasma carries out sealing-in between each layer, forms little valve on the chip by specific design.Secondly, build the fluid control module of being formed by the little valve on array micro syringe pump and the chip.Build cell and cultivate the microenvironment module.This module constitutes (as shown in Figure 3) by the cell culture chamber that can be fixed in microscope stage, gas control system, temperature humidity control system.Build cellular response real time monitoring signals module, mainly comprise automatically controlled fluorescence imaging platform, ccd signal acquisition system, data handling system three parts.At last, four modules are integrated into a complete system (as shown in Figure 4) by coupling arrangement
Keep the interior temperature of cell culture chamber at 37 ℃, CO
2Level is 5%, keeps higher relative saturation humidity (95%).With putting into cell culture chamber after the chip ultraviolet irradiation sterilization of making, cell culture chamber is fixed within the cellular response real time monitoring signals module.By the cell inoculation path injection hepatoma carcinoma cell HepG2 cell suspension (about 10 of syringe pump from chip center
7/ ml), spend the night.Inject support platinum glycosides by syringe pump by medicine 1 entrance, medicine 2 entrances inject carboplatin, and medicine 3 entrances inject mitomycin, by regulating the On/Off of each little valve on the chip, obtain variable concentrations and the different pharmaceutical combination of three kinds of medicines.After 24 hours, inoculate path by syringe pump from the cell of chip center and inject 4 μ g/ml Hoechst33342 and 2 μ g/ml Rh123, after 20 minutes, inject the PBS damping fluid and clean, the 2 μ g/mlPI that reinject adopt cellular response real time monitoring signals module to detect and take pictures.
Claims (5)
1. the horizontal micro-fluidic chip drug evaluation of fully integrated high-flux cell system is characterized in that: this system by drug evaluation with function chip module, drug evaluation with fluid control module, cell cultivate the microenvironment control module, four modules of the real-time monitoring modular of cell fluorescence response signal are formed.
2. according to the horizontal micro-fluidic chip drug evaluation of the described fully integrated high-flux cell of claim 1 system, it is characterized in that: described drug evaluation is made up of 1～10 identical structural unit with the function chip module, each structural unit can detect the effect of 1～10 kind of medicine, by accurate pump valve control, can form the combination of multiple drug concentration gradient or different pharmaceutical.
3. according to the horizontal micro-fluidic chip drug evaluation of the described fully integrated high-flux cell of claim 1 system, it is characterized in that: described drug evaluation fluid control module is the fluid control systems of micro-fluidic chip, is made up of the little valve/Micropump on array micro syringe pump and the chip.
4. according to the horizontal micro-fluidic chip drug evaluation of the described fully integrated high-flux cell of claim 1 system, it is characterized in that: described cell is cultivated the microenvironment control module and is made of the cell culture chamber that can be fixed in microscope stage, gas control system, temperature humidity control system.Chip places in this culturing room, keeps cell cultivation temperature, humidity and CO stable in it by a series of sensors and control module
2Gas concentration is achieved the monitoring in real time of the online cultivation of continuous cell and cell signal.
5. according to the horizontal micro-fluidic chip drug evaluation of the described fully integrated high-flux cell of claim 1 system, it is characterized in that: the real-time monitoring modular of described cell fluorescence response signal adopts the confocal fluorescent detection technique of automatically controlled scanning to realize that this module mainly comprises automatically controlled fluorescence imaging platform, ccd signal acquisition system, data handling system three parts.
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| CN105032512A (en) * | 2015-08-25 | 2015-11-11 | 辽宁中医药大学 | Integrated micro-fluidic chip for screening drug compatibility, as well as preparation method and application of integrated micro-fluidic chip |
| CN105241940A (en) * | 2015-08-05 | 2016-01-13 | 深圳大学 | Pharmacodynamic detection method and system thereof on the basis of dielectrophoresis force field |
| CN105670921A (en) * | 2016-03-09 | 2016-06-15 | 江汉大学 | Multichannel high-flux microfluidic chip for screening drugs for controlling plant diseases |
| CN105707010A (en) * | 2014-12-05 | 2016-06-29 | 中国科学院大连化学物理研究所 | Micro-valve-based dual-detection microfluidic chip and diabetic model construction method |
| CN106318866A (en) * | 2015-06-24 | 2017-01-11 | 中国科学院大连化学物理研究所 | Small multi-purpose cell culturing and testing device |
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