Summary of the invention
The object of this invention is to provide a kind of simple in structure, easy to operate, highly sensitive immune detection micro-fluidic chip fluorescent quantitation automatic detection device.The linear electric motors that utilize reaction to automatically perform in module drive the liquid storage Micropump on micro-fluidic chip automatically to complete complicated immune detection reaction.Meanwhile, utilize waste reaction solution to remove miniature electric resistance heater integrated in module to realize the contact spot heating of micro-fluidic chip, remove waste reaction solution by evaporation mode, improve the sample size of allowing of chip detection, improve thus detection sensitivity.Read in order to realize quantitative detection signal, based on total differential sampling Detection principle, carry out continuous sweep micro-fluidic chip by designing fine linear exciting light spot, simultaneously, carried out the fluorescence signal of continuous acquisition micro-fluidic chip by highly sensitive photodiode, thus, realize in micro-fluidic chip detection reaction region, the extraction that high resolving power (spacing of neighbouring sample point reaches μ m level) fluorescence signal intensity distributes, the built-in algorithm of recycling controller is extrapolated testing sample concentration, or number of cells.By the extraneous interface module in circuit module, can realize easily the function such as transmission, printout of data, be user-friendly to.Especially, the Bluetooth wireless communication module that can comprise by circuit module, is sent to medical diagnosis mechanism by testing result in real time by smart mobile phone, realizes remote diagnosis or the monitoring of epidemic disease distributed areas.
Particularly, the invention provides a kind of immune detection micro-fluidic chip fluorescent quantitation automatic detection device, comprise: reaction automatically performs module, waste reaction solution removes module, excitation source, exciting light focus module, phosphor collection module, linear movement console module, circuit module, and circuit module comprises: master controller, submaster controller and extraneous interface module.
In immune detection course of reaction, by the control of circuit module, reaction automatically performs the reaction liquid storage Micropump of one or more linear electric motors (mini drive permutation) driving micro-fluidic chip in module, and all kinds of reaction reagents are driven into relevant conversion zone, automatically completes detection reaction.Adopt electronic journey switch to realize linear electric motors motion positions.
In immune detection course of reaction, for the response sample amount of allowing by increase, reach the object that improves detection sensitivity, by the control of circuit module, allow waste reaction solution remove miniature electric resistance heater integrated in module to realize the contact spot heating of micro-fluidic chip, remove waste reaction solution by evaporation mode.The working temperature of miniature electric resistance heater detects by being fixed on its surperficial micro thermocouple temperature sensor, and feeds back to controller and realize the closed-loop control of ON/OFF pattern.
After immune response completes, testing result reads in process, the original hot spot that excites that excitation source produces is assembled for the linear hot spot that excites through exciting light focus module, in order to improve the spatial resolution of detection signal, the accurately physical boundary in recognition detection signal distributions region, improve detection accuracy, the width of line spot should enough carefully (as: 50-250 μ m), simultaneously, distribute in order to obtain complete micro-fluidic chip fluorescence signal, the length of line spot should approach with the cross-sectional width in micro-fluidic chip two dimension rule detection region.In order to improve the scanning accuracy of detection of micro-fluidic immune detection chip, realize accurately and quantitatively detecting, utilize the segmentation control technology of stepper motor, pass through belt drive system, realize at the uniform velocity micro-fluidic chip linear movement, make the physical location spacing of neighbouring sample point reach μ m rank and improved the resolution that scanning detects.The fluorescence producing on micro-fluidic chip enters phosphor collection module, is converged by lens wherein, finally arrives highly sensitive photoelectric conversion module.Submaster controller continuous acquisition photoelectric conversion signal, and send it to master controller, detect the fluorescence signal intensity distributed intelligence obtaining based on scanning, extrapolate quantitative testing result (sample concentration or number of cells) according to built-in analytical algorithm and mathematical model.
In order to reduce installation cost, adopt LED, and nonideal pointolite---laser, therefore, in order to improve detection resolution, the present invention need to design unique exciting light focus module, by cooperatively interacting between multiple condenser lenses, cylindrical mirror, obtains scanning the linear hot spot that excites of micro-fluidic chip.
By the Bluetooth wireless communication module in extraneous interface module, can realize the exchanges data between this pick-up unit and PC, smart mobile phone.On the one hand, in can examinations process, the control to pick-up unit of PC or smart mobile phone, on the other hand, testing result can be transferred to PC, or smart mobile phone, then send to all kinds of medical diagnosis mechanism by mobile network communication technology such as the GPRS network of Internet network or mobile phone, realizes intelligent, informationalized remote diagnosis or the monitoring of epidemic disease distributed areas.
Preferably, in course of reaction, the liquid storage Micropump that reaction automatically performs on module drive micro-fluidic chip completes immune detection reaction automatically.React the linear electric motors of automatic driver module and realize positioning control by electronic journey switch.
Preferably, in course of reaction, waste reaction solution removes microheater integrated in module and removes sample waste liquid by evaporation mode, and the application of sample amount of allowing that increases response sample improves detection sensitivity.The temperature of microheater detects and feeds back to submaster controller through its surperficial micro thermocouple temperature sensor and realizes ON/OFF closed-loop control.
Preferably, exciting light projects on micro-fluidic chip after exciting light focus module, and the fluorescence exciting on micro-fluidic chip receives through phosphor collection module, then is converted to electric signal by photoelectric conversion module (photodiode), finally by circuit module is processed.
Preferably, the relative position of exciting light focus module and linear movement console module can regulate, and the relative position of phosphor collection module and linear movement console module also can regulate, and the micro-fluidic chip scanning that can realize thus high flexibility ratio detects.
Preferably, this device also comprises the high-pass filter (22) being arranged between phosphor collection module (5) and photoelectric conversion module (6), high-pass filter (22) filter wavelength is greater than and excites light wavelength, and be arranged on the bandpass filter (21) between excitation source (1) and exciting light focus module (2), after bandpass filter (21) filters, wavelength, at excitation wavelength ± 35nm, is respectively used to the fluorescence to entering photoelectric conversion module (6) and the exciting light that enters exciting light focus module (2) is filtered.
Preferably, linear movement console module comprise stepper motor, driving belt and with the fixing draw-in groove of driving belt, wherein draw-in groove is for placing detected micro-fluidic chip, when stepper motor rotates under the driving of submaster controller, drive belt, draw-in groove and micro-fluidic chip wherein to move linearly.
Preferably, master controller is used for analysis detecting data, and according to built-in mathematical model, quantitatively calculates the concentration value that detects sample, or the number of cells of testing sample.
Preferably, light source is LED, and submaster controller can be controlled the on off state of excitation source, and can digitizing constant current source size be set and regulates the brightness of light source, obtains optimum detection signal-to-noise ratio.
Preferably, in circuit module, comprise Bluetooth wireless communication module, for realizing the exchanges data between this pick-up unit and PC, smart mobile phone.
Embodiment
Below in conjunction with the accompanying drawing in the embodiment of the present invention, the technical scheme in the embodiment of the present invention is clearly and completely described.Obviously, described embodiment is only the present invention's part embodiment, instead of whole embodiment.In fact be only illustrative to the description of at least one exemplary embodiment below, never as any restriction to the present invention and application or use.Based on the embodiment in the present invention, those of ordinary skill in the art, not making all other embodiment that obtain under creative work prerequisite, belong to the scope of protection of the invention.
Unless illustrate in addition, otherwise the parts of setting forth in these embodiments and positioned opposite, numeral expression formula and the numerical value of step not limited the scope of the invention.
Meanwhile, should be clear and definite, for convenience of description, the size of the various piece shown in accompanying drawing is not to draw according to actual proportionate relationship.
May not discuss in detail for the known technology of person of ordinary skill in the relevant, method and apparatus, but in suitable situation, described technology, method and apparatus should be regarded as authorizing a part for instructions.
In all examples with discussing shown here, it is exemplary that any occurrence should be construed as merely, instead of as restriction.Therefore, other example of exemplary embodiment can have different values.
It should be noted that: in similar label and letter accompanying drawing below, represent similar terms, therefore, once be defined in an a certain Xiang Yi accompanying drawing, in accompanying drawing subsequently, do not need it to be further discussed.
Fig. 1 is the schematic diagram of an embodiment of immune detection micro-fluidic chip fluorescent quantitation automatic detection device of the present invention.As shown in Figure 1, immune detection micro-fluidic chip fluorescent quantitation automatic detection device comprises that excitation source (1), exciting light focus module (2), linear movement console module (3), phosphor collection module (5), photoelectric conversion module (6), circuit module (7), waste reaction solution remove module (8), reaction automatically performs module (9), and circuit module (7) comprises master controller (71), submaster controller (72) and extraneous interface module (73).
In course of reaction, by the control of circuit module 7, reaction automatically performs the reaction liquid storage Micropump of one or more linear electric motors (mini drive permutation) driving micro-fluidic chip in module 9, and all kinds of reaction reagents are driven into relevant conversion zone, automatically completes detection reaction.
In course of reaction, by the control of circuit module 7, allow waste reaction solution remove miniature electric resistance heater integrated in module 8 to realize the contact spot heating of micro-fluidic chip, remove waste reaction solution by evaporation mode, improve the application of sample amount of allowing of response sample.
Testing result reads in process, excitation source 1 produces exciting light, exciting light focuses on the micro-fluidic chip 4 that is positioned at linear movement platform 3 after by exciting light focus module 2, linear movement platform 3 drives micro-fluidic chip 4 to move, and has realized the scanning of exciting light to micro-fluidic chip two dimension rule detection region.
In micro-fluidic chip 4 motion processes, on micro-fluidic chip 4, in two-dimentional rule detection region, the fluorescent marker of ad-hoc location is excited by linear exciting light, the fluorescence producing is collected by phosphor collection module 5, finally arrives highly sensitive photoelectric conversion module 6, and light signal is converted to electric signal.In scanning testing process, the electric signal that submaster controller 72 continuous acquisition photoelectric conversion modules 6 obtain, and be sent to master controller 71, master controller 71 utilizes built-in data processing algorithm to carry out data analysis, and accurately calculate testing sample concentration, or target cell number in sample.The Bluetooth wireless communication module comprising by extraneous interface module 73, testing result can be transferred to PC, or smart mobile phone, send to all kinds of medical diagnosis mechanism by mobile network communication technology such as the GPRS network of Internet network or mobile phone again, realize intelligent, informationalized remote diagnosis or the monitoring of epidemic disease distributed areas.
The immune detection micro-fluidic chip fluorescent quantitation automatic detection device providing based on the above embodiment of the present invention, automatically perform module by reaction and automatically complete immune detection reaction, remove module by waste reaction solution and increase the immunoreactive application of sample amount of allowing, finally realize the extraction on chip detection region fluorescence signal intensity and border by one dimension line spot scanning detecting pattern, and utilize built-in algorithm to extrapolate testing sample concentration, or number of cells.On the basis that the present invention detects at fluorescent quantitation, realized the immunoreactive automation mechanized operation of micro-fluidic chip, the response sample amount of allowing by increase chip has improved detection sensitivity.
Preferably, reaction automatically performs module 9 and adopts linear electric motors to drive the liquid storage Micropump of micro-fluidic chip, and linear electric motors are realized position probing by electronic journey switch, have realized immunoreactive automation mechanized operation on micro-fluidic chip.
Preferably, waste reaction solution remove module 8 integrated microheater remove waste reaction solution by evaporation pattern, working temperature detects and feeds back to the submaster controller 71 in circuit module 7 by being fixed on its surperficial micro thermocouple temperature sensor, realizes the closed-loop control based on ON/OFF pattern.
Preferably, excitation source 1 is LED(LightEmittingDiode, light emitting diode).That the excitation wavelength that in a preferred embodiment, light source 1 provides is selected is 365nm.
Preferably, producing glimmering light wavelength is 615nm.
Preferably, the photoelectric conversion module adopting is near photodiode (sensitivity: 145V/ μ W, wavelength 600nm).
Preferably, controlled the on off state of excitation source 1 by submaster controller 72, and can digitizing constant current source size be set and regulates the brightness of light source, obtain optimum testing result.
Preferably, linear movement console module 3 comprise stepper motor, driving belt and with the fixing draw-in groove of driving belt, wherein draw-in groove is used for placing micro-fluidic chip 4, when stepper motor rotates under the driving of submaster controller 72, drive belt, draw-in groove and micro-fluidic chip wherein 4 to move linearly, simultaneously, under the effect of linear exciting light spot, complete the fluorescence signal collection in micro-fluidic chip two dimension rule detection region, finally obtaining complete fluorescence signal intensity distributes, and extrapolate thus testing sample concentration, or target cell number in sample.
Preferably, exciting light enters into exciting light focus module 2, first converge to a bit through the condenser lens 201 in exciting light focus module 2, by cylindrical mirror 202, make exciting light be converted to the line spot of scanning micro-fluidic chip, this line spot projects in the two-dimentional rule detection region of linear stage micro-fluidic chip fluorescence excitation label on ad-hoc location.
Preferably, extraneous interface module 73 comprises Bluetooth wireless communication module, for realizing the exchanges data between this pick-up unit and PC, smart mobile phone.
In one embodiment, extraneous interface module 73 can be realized and extraneous communicating by letter.
Preferably, extraneous interface module 73 can comprise for carrying out the USB(Universal Serial Bus of data transmission, USB (universal serial bus) with external computing machine) interface.
Preferably, extraneous interface module 73 can comprise the printer interface for external printer, external printer printable go out final testing result.
Preferably, extraneous interface module 73 can comprise for carrying out the network port of real-time Communication for Power with external system.
Preferably, extraneous interface module 73 can comprise the display interface for external-connection displayer.
Fig. 2 is the schematic diagram of another embodiment of immune detection micro-fluidic chip fluorescent quantitation automatic detection device of the present invention.Compared with embodiment illustrated in fig. 1, in embodiment illustrated in fig. 2, also comprise the bandpass filter 21 being arranged between excitation source 1 and exciting light focus module 2, for the exciting light that enters exciting light focus module 2 is filtered.
The effect that bandpass filter 21 is set is only to allow the light that meets specific wavelength condition to pass through.Preferably, it is 320-390nm that bandpass filter 21 allows the light wavelength scope of passing through, and the exciting light that is mainly used in allowing excitation source 1 to produce passes through.
Preferably, in embodiment illustrated in fig. 2, also comprise the high-pass filter 22 being arranged between phosphor collection module 5 and photoelectric conversion module 6, for the fluorescence that enters photoelectric conversion module 6 is filtered.
The effect that high-pass filter 22 is set is only to allow the light that meets specific wavelength condition to pass through.Preferably, high-pass filter 22 allows the light wavelength scope of passing through for being greater than 580nm, and the exciting light that is mainly used in avoiding light source 1 to produce enters phosphor collection module 5.
As an example of Fig. 2 example, the present invention will be described below.
In immune detection course of reaction, by the control of circuit module, reaction automatically performs the liquid storage Micropump of the linear electric motors driving micro-fluidic chip in module, automatically completes immune detection reaction.
In immune detection course of reaction, by the control of circuit module, the microheater that waste reaction solution removes in module has removed waste reaction solution by evaporation mode, has increased the detection sample pipetting volume amount of allowing, has improved detection sensitivity.
Testing result reads in process, and excitation source 1 produces exciting light, and exciting light, by after the filtration of bandpass filter 21, enters exciting light focus module 2, to only allow the light that meets respective wavelength requirement to enter exciting light focus module 2.Exciting light converges on the micro-fluidic chip 4 that is placed on linear stage 3 after by exciting light focus module, and linear stage 3 drives micro-fluidic chip 4 to move, and exciting light can scan detection to whole micro-fluidic chip thus.
Fluorescent marker on micro-fluidic chip 4 is excited after irradiation and produces fluorescence, fluorescence arrives high-pass filter 22 by the lens in phosphor collection module 5, after high-pass filter 22 filters, arrive photoelectric conversion module 6, thereby only allow to meet light (exciting the fluorescence of generation) the arrival photoelectric conversion module 6 that respective wavelength requires.
Light signal is converted to electric signal after photoelectric conversion module 6, in collected submaster controller 72, submaster controller 72 is sent into signal in master controller 71 and is analyzed, and the testing result of final quantitative (detect sample concentration or detect target cell number in sample).The Bluetooth wireless communication module comprising by extraneous interface module 73, testing result can be transferred to PC, or smart mobile phone, send to all kinds of medical diagnosis mechanism by mobile network communication technology such as the GPRS network of Internet network or mobile phone again, realize intelligent, informationalized remote diagnosis or the monitoring of epidemic disease distributed areas.
Fig. 3 is the inner structure schematic diagram of an embodiment of exciting light focus module of the present invention.Exciting light focus module 2 comprises condenser lens 201 and cylindrical mirror 202, and condenser lens 201 is for exciting light being converged to a luminous point, and cylindrical mirror 202 is converted to this focal spot the line spot of scanning micro-fluidic chip again.
By implementing the present invention, can obtain following beneficial effect:
1, in the present invention, react and automatically perform module by introducing, drive the liquid storage Micropump of micro-fluidic chip, automatically complicated immunoreactive multiple operation stepss have been completed, as detect sample quantitative sample injection, and discharge washing lotion, label etc., realize the automation mechanized operation of immune detection reaction on micro-fluidic chip, improve detection efficiency and repeatability, avoided makeing mistakes of manual operation.
2, in the present invention, remove module by introducing waste reaction solution, adopt evaporation mode automatically to remove waste reaction solution, increase thus the detection sample pipetting volume amount of allowing, in detection reaction process, in bulk sample, the enrichment effect of label to be measured, has improved detection sensitivity.
3, in the present invention, exciting light can be converted to the line spot of scanning micro-fluidic chip after exciting light focus module.Accurate motion control by submaster controller to stepper motor, realize the linear movement of micro-fluidic chip, under the effect of linear exciting light spot, complete the fluorescence signal collection in micro-fluidic chip two dimension rule detection region, finally having obtained complete fluorescence signal intensity distributes, and extrapolate thus testing sample concentration, or detect target cell number in sample.The present invention, in having guaranteed accuracy in detection and sensitivity, has adopted the one-dimensional scanning detection mode that excites hot spot based on linear, has significantly reduced system complexity, has simplified testing process.
One of ordinary skill in the art will appreciate that all or part of step that realizes above-described embodiment can complete by hardware, also can control relevant hardware by program completes, described program can be stored in a kind of computer-readable recording medium, the above-mentioned storage medium of mentioning can be ROM (read-only memory), disk or CD etc.
Description of the invention provides for example with for the purpose of describing, and is not exhaustively or limit the invention to disclosed form.Many modifications and variations are obvious for the ordinary skill in the art.Selecting and describing embodiment is for better explanation principle of the present invention and practical application, thereby and makes those of ordinary skill in the art can understand the present invention's design to be suitable for the various embodiment with various amendments of special-purpose.