CN103529006A - Portable fluorescence detection device based on micro-fluidic chip and detection method thereof - Google Patents
Portable fluorescence detection device based on micro-fluidic chip and detection method thereof Download PDFInfo
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- CN103529006A CN103529006A CN201310491678.XA CN201310491678A CN103529006A CN 103529006 A CN103529006 A CN 103529006A CN 201310491678 A CN201310491678 A CN 201310491678A CN 103529006 A CN103529006 A CN 103529006A
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Abstract
The invention discloses a portable fluorescence detection device based on a micro-fluidic chip and a detection method thereof. The device comprises a light source spotlight component, a micro-fluidic chip loading platform, a micro-fluidic chip, a fluorescence collecting component, a light detection device, a signal processing component and a display component, wherein the micro-fluidic chip is fixed in the micro-fluidic chip loading platform; the light source spotlight component is positioned on the micro-fluidic chip loading platform; the fluorescence collecting component is fixed below the micro-fluidic chip loading platform; the light detection device is fixed at the bottom of the fluorescence collecting component; the light detection device, the signal processing component and the display component are serially connected through signal cables in sequence. The portable fluorescence detection device is high in detection sensitivity, can detect fluorescence signals of fluorescence grains of 2.06 micrometers, and has unique advantages in detection on weak fluorescence signals. The portable fluorescence detection device has the advantages of small size, light weight, portability and low price, can be held by hand, and is especially applicable to on-site detection.
Description
Technical field
The present invention relates to a kind of detection technique of fluorescent samples, particularly a kind of Portable fluorescence pick-up unit and detection method thereof based on micro-fluidic chip.
Background technology
At present, flow cytometer (flowcytometry, FCM) be the instrument that the mobile cell of high-speed straight-line or biological particle are carried out to quantitative determination and analysis, also be applicable to the determination and analysis of fluorescent samples, mainly comprise the flow technique of sample, the counting of cell and sorting technology, computing machine is to the collection of data and analytical technology etc.Flow cytometer be take flow cytometry as theoretical foundation, is the crystallization of the subject knowledge integrated applications such as fluid mechanics, laser technology, electronic engineering, molecular immunology, cell fluorescence and computing machine.Be characterized in: fast, the tested colony of measuring speed is large, can carry out measuring multiple parameters, at biological, medical domain, is widely used.But this instrument is bulky, expensive, can not be integrated portable, be also unfavorable for widespread use.
For above shortcoming, people have proposed a kind of portable flow cytometer based on micro-fluidic chip, micro-fluidic chip (microfluidicchip) technology is integrated into the unit such as pre-service, reaction, separation and detection in one single chip, there is the features such as integrated level is high, volume is little, represented the direction of microminiaturized Instrument Development.The detection technique adopting is optical detection, and concrete light source is LED light source, along with the improvement of maturation and the performance of technology, the LED that high power, short wavelength, monochromaticity can be provided and have good stability.Adopt LED can reduce costs as excitation source, reduce volume and be more conducive to portable integrated.Light detecting device and display device are also selected microminiaturized equipment.It is high that portable flow cytometer based on micro-fluidic chip has integrated level, and cost is low, is adapted to the advantage of Site Detection.But it is not high that the shortcoming of this instrument is detection sensitivity, the sample very faint for fluorescence cannot carry out determination and analysis, such as being that 3.75 μ m and the following fluorescent particle of 3.75 μ m cannot detect to size.
Summary of the invention
The problems referred to above that exist for solving prior art, the present invention will design a kind ofly to be had higher detection sensitivity and can detect more Portable fluorescence pick-up unit and the detection method thereof based on micro-fluidic chip of fluorescent signals.
To achieve these goals, technical scheme of the present invention is as follows:
A Portable fluorescence pick-up unit for micro-fluidic chip, comprises light source concentrating component, micro-fluidic chip carrying platform, micro-fluidic chip, phosphor collection assembly, light detecting device, signal processing component and display module; Described micro-fluidic chip is fixed in micro-fluidic chip carrying platform, described light source concentrating component is positioned on micro-fluidic chip carrying platform, described phosphor collection assembly be fixed on micro-fluidic chip carrying platform below, described light detecting device is fixed on the bottom of phosphor collection assembly, and described light detecting device, signal processing component and display module are connected in series by signal cable order;
Described light source concentrating component comprises LED light source, collector lens A, optical filter A and collector lens B, described LED light source below is followed successively by collector lens A, optical filter A and collector lens B, described LED light source band collimating apparatus, the slightly larger in diameter of described collector lens A is in the diameter of LED light source;
Described micro-fluidic chip is comprised of substrate and cover plate, and the two is sealed to and forms together microchannel, and described substrate is dimethyl silicone polymer or polymethylmethacrylate, and described cover plate is glass sheet;
Described phosphor collection assembly is included in detection slit, optical filter B, the collector lens C on micro-fluidic carrying platform, described detection slit below is provided with optical filter B and collector lens C successively, described detection slit is the region that fluorescence sees through, and the diameter of described collector lens C is larger than the diameter of light detecting device;
Described signal processing component adopts phase shift difference channel, comprises phase-shift circuit, difference channel and filter amplification circuit; Input signal is divided into the positive input terminal that two-way ，Yi road directly enters into difference channel, and another road enters the negative input end of difference channel after the very short time of phase-shift circuit time delay, then after amplifying circuit, obtains output signal after filtering.
A detection method for the Portable fluorescence pick-up unit of micro-fluidic chip, comprises the following steps:
A, collector lens A collect the directional light shooting out from LED light source collimating apparatus, then converge to collector lens B upper, by regulating the distance between collector lens A collector lens B, obtain suitable focal length, before optical filter A is placed on excitation light emission, provide monochromaticity better exciting light;
The excitation beam of B, the transmitting of light source concentrating component is radiated at and detects on slit, sample in the microchannel of micro-fluidic chip is flowed through while detecting slit, is inspired fluorescence, detects slit whole fluorescence is passed through, and block a large amount of exciting lights, play the effect of spatial filtering; Optical filter B, by seeing through the further filtering of light that detects slit, is finally irradiated on collector lens C;
The fluorescence that C, light detecting device converge collector lens C is converted into corresponding electric signal;
D, signal processing component carry out filter amplifying processing to electric signal, suppress noise amplifying signal, improve signal to noise ratio (S/N ratio);
E, display module show the signal after signal processing component is processed.
Compared with prior art, the present invention has following beneficial effect:
1, the present invention is by adopting LED as excitation source, micro-fluidic chip is as the microfluidic platform of fluoroscopic examination, relevant Photoelectric Detection equipment and display device also can adopt the version of small volume, make checkout equipment have volume little, lightweight, be easy to carry, low price, can carry out hand-held, for the advantage of Site Detection.
2, the present invention is by adopting the method for optical concentration, spatial filtering and phase shift differential filtering amplifying circuit, improved detection sensitivity, size can be detected is the fluorescence signal of 2.06 μ m fluorescent grains, in fluorescent signals context of detection, has unique advantage.
Accompanying drawing explanation
6, the total accompanying drawing of the present invention, wherein:
Fig. 1 is the structural representation of portable device with high sensitivity for detecting fluorescence;
Fig. 2 is the structural representation of light source concentrating component;
Fig. 3 is the structure side view of phosphor collection assembly;
Fig. 4 is the structure vertical view of phosphor collection assembly;
Fig. 5 is the structural representation of phase shift difference channel;
Fig. 6 is the result figure that detects the standard fluorescence particle of 2.06 μ m.
In figure: 1, light source concentrating component, 2, micro-fluidic chip carrying platform, 3, micro-fluidic chip, 4, phosphor collection assembly, 5, light detecting device, 6, signal processing component, 7, display module, 11, LED light source, 12, collector lens A, 13, optical filter A, 14, collector lens B, 31, microchannel, 41, detect slit, 42, optical filter B, 43, collector lens C.
Embodiment
Below in conjunction with accompanying drawing and example, the invention will be further described.Fig. 1 shows one-piece construction block diagram of the present invention, and as seen from Figure 1, the present invention is comprised of light source concentrating component 1, micro-fluidic chip carrying platform 2, micro-fluidic chip 3, phosphor collection assembly 4, light detecting device 5, signal processing component 6 and display module 7.As seen from Figure 2, light source concentrating component 1 is by the LED light source 11 with collimating apparatus, collector lens A12, collector lens B14 and optical filter A13, wherein LED light source 11 is ethereal blue radiant, collector lens A12 immediately below, diameter is 23.9mm, than the slightly larger in diameter of LED light source 11, focal length is 14mm, the parallel beam of LED light source 11 collimations is converged, be irradiated to collector lens B14 below, diameter is 12.7mm, focal length is 13mm, the object that adds collector lens B14 is mainly to allow focal length shorten save space, make the less light intensity of luminous point stronger, the selection of collector lens is not unique, as long as can meet above-mentioned optically focused requirement, distance between collector lens A12 and collector lens B14 is 5mm, in order to save space, the thick 3mm of being about of optical filter A13 is placed between collector lens A12 and collector lens B14, LED light source 11 is ethereal blue light, so the centre wavelength of optical filter is 480nm, bandwidth is 30nm, make exciting light purer.Fig. 3 and 4 is respectively side view and the vertical view of phosphor collection assembly 4, figure from two different visual angles, phosphor collection assembly 4 comprises detection slit 41, optical filter B42, collector lens C43, in vertical view, flow through and detect slit 41 in microchannel 31, in side view, photoelectricity testing part 5 is fixed on the bottom of phosphor collection assembly 4, the optically focused light beam irradiates of light source concentrating component 1 transmitting is detecting on slit 41, in microchannel 31, there is sample to flow through while detecting slit 41, sample is inspired fluorescence, detecting slit 41 enters fluorescence, blocked a large amount of exciting lights simultaneously, play the effect of spatial filtering, the fluorescence entering and veiling glare be the filtration of mating plate B42 after filtration again, reduce veiling glare to the interference detecting, through converging of collector lens C43, the most of fluorescence seeing through can be converged on the surveyed area of light detecting device 5, fluorescence is converted into electric signal, light detecting device 5 is photodiodes, be fixed on the bottom of phosphor collection assembly 4.Because the fluorescence of being sent out is green glow, so the centre wavelength that optical filter B42 selects is 530nm, bandwidth is 40nm.The diameter that collector lens C43 selects is that 5mm, focal length are 4mm.Distance between light detecting device 5 and collector lens C43 is 1mm, guarantees that more fluorescence can converge on the surveyed area of light detecting device 5.Light source concentrating component 1 and phosphor collection assembly 4 are only to allow Whole Equipment volume slightly increase, but are to improve a lot in the detection sensitivity of system.As seen from Figure 5, by the processed flow process of photoelectricity testing part electric signal out, one road signal enters into the positive input of difference channel, one tunnel enters into the negative input of difference channel after by phase shift time delay, then carry out difference, phase-shift circuit is RC filtering circuit, that differential amplifier is used is AD620, phase shift difference can be by much noise filtering and stick signal, from the amplifying circuit processing after filtering again of AD620 signal out, filter amplification circuit is second order Butterworth LPF, according to the frequency 1hz of signal, setting cutoff frequency is 5hz, amplifies 2500 times.Phase shift difference after filter and amplification below, can greatly suppress noise again, and signal amplifies, and significantly improves signal to noise ratio (S/N ratio), and the antijamming capability of circuit is strengthened.
Adopting the present invention is 2.06 μ m time shutter to be 13.23s standard fluorescence particle detects to size, sample thief joins in micro-fluidic chip 3, fluorescent grain flows in microchannel 31, while detecting slit 41 when flowing through, fluorescent grain is by the beam excitation of light source concentrating component 1 transmitting, produce fluorescence, by phosphor collection assembly 4 by phosphor collection to the surveyed area of light detecting device 5, light detecting device 5 is converted into electric signal by corresponding light signal, pass through again the filter amplifying processing of signal processing component 6, be input to display module 7, the signal of high s/n ratio shows the most at last.The amplitude of corresponding signal is exactly the power of particle fluorescence.Fig. 6, for size is the testing result of 2.06 μ m fluorescent grains, as can be seen from the figure can detect the fluorescent grain that size is 2.06 μ m, has illustrated that the present invention has very high detection sensitivity.
Claims (2)
1. the Portable fluorescence pick-up unit based on micro-fluidic chip, is characterized in that: comprise light source concentrating component (1), micro-fluidic chip carrying platform (2), micro-fluidic chip (3), phosphor collection assembly (4), light detecting device (5), signal processing component (6) and display module (7); Described micro-fluidic chip (3) is fixed in micro-fluidic chip carrying platform (2), described light source concentrating component (1) is positioned on micro-fluidic chip carrying platform (2), described phosphor collection assembly (4) be fixed on micro-fluidic chip carrying platform (2) below, described light detecting device (5) is fixed on the bottom of phosphor collection assembly (4), and described light detecting device (5), signal processing component (6) and display module (7) are connected in series by signal cable order;
Described light source concentrating component (1) comprises LED light source (11), collector lens A(12), optical filter A(13) and collector lens B(14), described LED light source (11) below is followed successively by collector lens A(12), optical filter A(13) and collector lens B(14), described LED light source (11) band collimating apparatus, described collector lens A(12) slightly larger in diameter is in the diameter of LED light source (11);
Described micro-fluidic chip (3) is comprised of substrate and cover plate, and the two is sealed to and forms together microchannel (31), and described substrate is dimethyl silicone polymer or polymethylmethacrylate, and described cover plate is glass sheet;
Described phosphor collection assembly (4) is included in detection slit (41), the optical filter B(42 on micro-fluidic carrying platform), collector lens C(43), described detection slit (41) below is provided with optical filter B(42 successively) and collector lens C(43), the region that described detection slit (41) sees through for fluorescence, described collector lens C(43) diameter is larger than the diameter of light detecting device (5);
Described signal processing component (6) adopts phase shift difference channel, comprises phase-shift circuit, difference channel and filter amplification circuit; Input signal is divided into the positive input terminal that two-way ，Yi road directly enters into difference channel, and another road enters the negative input end of difference channel after the very short time of phase-shift circuit time delay, then after amplifying circuit, obtains output signal after filtering.
2. a detection method for the Portable fluorescence pick-up unit based on micro-fluidic chip, is characterized in that: comprise the following steps:
A, collector lens A(12) collect the directional light shoot out from LED light source (11) collimating apparatus, then converge to collector lens B(14) on, by regulating collector lens A(12) collector lens B(14) between distance, obtain suitable focal length, optical filter A(13), before being placed on excitation light emission, provide monochromaticity better exciting light;
The excitation beam of B, light source concentrating component (1) transmitting is radiated at and detects on slit (41), sample in the microchannel (31) of micro-fluidic chip (3) is flowed through while detecting slit (41), be inspired fluorescence, detecting slit (41) passes through whole fluorescence, and block a large amount of exciting lights, play the effect of spatial filtering; Optical filter B(42) will, through the further filtering of light that detects slit (41), finally be irradiated to collector lens C(43) on;
C, light detecting device (5) are by collector lens C(43) fluorescence that converges is converted into corresponding electric signal;
D, signal processing component (6) carry out filter amplifying processing to electric signal, suppress noise amplifying signal, improve signal to noise ratio (S/N ratio);
E, display module (7) show the signal after signal processing component (6) is processed.
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