CN112903793A - Multi-parameter rapid detection device and method for water body - Google Patents

Multi-parameter rapid detection device and method for water body Download PDF

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CN112903793A
CN112903793A CN202110088299.0A CN202110088299A CN112903793A CN 112903793 A CN112903793 A CN 112903793A CN 202110088299 A CN202110088299 A CN 202110088299A CN 112903793 A CN112903793 A CN 112903793A
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CN112903793B (en
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田扬超
刘刚
熊瑛
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University of Science and Technology of China USTC
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Abstract

The invention relates to a multi-parameter rapid detection device and a detection method for a water body, wherein the device comprises a micro-fluidic chip module, and the micro-fluidic chip module comprises a dielectrophoresis bacteria sorting chip module, an impedance bacteria testing chip module, an extraction fluorescence testing chip module and an electrophoresis ion chip module; the microfluidic chip module is provided with four inlets, wherein the inlet I is used as a sample inlet, and the inlet II, the inlet III and the inlet IV are respectively used as a buffer solution inlet I, a buffer solution inlet II and a buffer solution inlet III; the device adopts a micro-fluidic technology, realizes the accurate flow direction control of liquid through a micro-channel and a micro-valve, highly integrates a plurality of pretreatment and detection modules on a small plastic sheet, and has higher integration level and high-efficiency mass transfer and detection speed. The whole set of device has stronger portability and higher detection speed and timeliness. A plurality of modules are integrated on the same chip, so that multi-parameter joint detection is realized, and only one sample needs to be input, so that the detection accuracy is higher.

Description

Multi-parameter rapid detection device and method for water body
Technical Field
The invention relates to the technical field of environmental detection, in particular to a multi-parameter rapid detection device and a multi-parameter rapid detection method for a water body.
Background
The areas of rivers, lakes and seas and other water areas in China are wide, and the quality of the water body has important influence on the production and life of people. The detection of water quality is the primary step of water body measurement. At present, two types of methods are used for measuring the quality of the traditional water body, one type of method is to collect a water sample on site and bring the water sample back to an analysis and detection laboratory for assay and analysis in a short time, traditional laboratory chemical measuring methods such as an atomic absorption method and an ultraviolet colorimetric method are mainly adopted, although the method has higher precision, the process is complicated, and a large amount of manpower, material resources and financial resources are consumed. The other method is to establish a small water quality monitoring station near the site, realize real-time water quality detection by establishing an automatic pretreatment pool and large automatic detection equipment, reduce links such as a large amount of manpower sampling and sample storage, but need site construction and consume a large amount of material resources and financial resources. Meanwhile, the methods adopted by the large-scale detection equipment need a large amount of chemical reagents, and cause secondary pollution to the environment. Therefore, establishing an analytical method which can be used for rapidly and accurately detecting water quality on site has important significance for environmental protection in China.
The micro-fluidic technology is to realize the control and transmission of micro liquid by constructing a micron-sized micro channel, and has the advantages of small sample demand, high mass transfer speed, small volume, easy portability, easy integration with detection means such as electro-optics and the like. The microchip manufactured by the micro-fluidic technology has extremely low material cost, can package a trace reaction reagent in advance, and has no secondary pollution to the environment due to the material and the trace reagent.
Spectrophotometry is an important component of spectroscopy, and is a method for qualitatively and quantitatively analyzing a substance to be detected by measuring the absorbance or luminous intensity of the substance at a specific wavelength or within a certain wavelength range, so that interference is avoided and the sensitivity is improved. The non-contact conductivity detection is applied to the detection of complex samples due to the advantages of high sensitivity, low cost, stable performance and the like. The electrophoresis separation technology has become a mainstream analysis tool for substance analysis, can sequentially separate inorganic ions in a complex sample so as to perform quantitative detection, and has the advantages of high detection speed, high precision, less sample consumption and the like. Compared with the traditional detection method, the electrophoresis-non-contact conductivity detection technology can detect several indexes within several minutes at the same time through one experiment, and has quicker detection and lower cost.
In recent years, with the rapid development of the microfluidic technology, the microfluidic technology is combined with the optical electrical detection technology, and the portable water quality multi-parameter rapid detection of the microchip is hopeful to be realized. However, no relevant devices carrying this technology are currently reported. Therefore, it is necessary to develop a new device and control method for realizing rapid and accurate determination of multiple parameters of water quality, thereby monitoring water quality. Considering that the cost of multi-section multi-technology detection and multi-index detection can be obviously improved, the cost of the integrated high-integration micro-fluidic chip can be obviously reduced.
Disclosure of Invention
The invention provides a multi-parameter rapid detection device and a multi-parameter rapid detection method for a water body, which can solve the technical problems.
In order to achieve the purpose, the invention adopts the following technical scheme:
a multi-parameter rapid detection device for a water body comprises a micro-fluidic chip module, wherein the micro-fluidic chip module comprises a dielectrophoresis bacteria sorting chip module, an impedance bacteria testing chip module, an extraction fluorescence testing chip module and an electrophoresis ion chip module;
the microfluidic chip module is provided with four inlets, wherein the inlet I is used as a sample inlet, and the inlet II, the inlet III and the inlet IV are respectively used as a buffer solution inlet I, a buffer solution inlet II and a buffer solution inlet III;
the dielectrophoresis bacteria sorting chip module is respectively communicated with the first inlet and the sample waste liquid outlet through the micro-channel;
the dielectrophoresis bacteria sorting chip module is also communicated with the second inlet through a micro-channel;
the dielectrophoresis bacteria sorting chip module is communicated with the impedance bacteria test chip module through a micro-channel, and an on-chip micro electromagnetic valve I is arranged between the dielectrophoresis bacteria sorting chip module and the impedance bacteria test chip module;
the dielectrophoresis bacteria sorting chip module is respectively communicated with the extraction fluorescence test chip module and the electrophoresis ion chip module through microchannels by an on-chip micro electromagnetic valve II;
the extraction fluorescence test chip module is also communicated with the inlet III through a micro-channel, and the electrophoresis ion chip module is communicated with the inlet IV through a micro-channel;
the impedance bacteria test chip module, the extraction fluorescence test chip module and the electrophoresis ion chip module are respectively communicated with the first liquid outlet, the second liquid outlet and the third liquid outlet in a one-to-one correspondence manner through microchannels;
the device further comprises an upper computer, and the first micro electromagnetic valve and the second micro electromagnetic valve are respectively connected with the upper computer.
Furthermore, a pipe of the sample bottle is connected to an inlet of the quantitative electromagnetic pump, and an outlet of the quantitative electromagnetic pump is connected with an inlet of the first flowmeter;
an outlet of the first flowmeter is connected with a sample inlet of the micro-fluidic chip, a sample waste liquid outlet of the micro-fluidic chip is connected to an inlet of the second flowmeter, and an outlet of the second flowmeter is connected to a waste liquid storage place;
the buffer solution I is connected to the inlet II through the rotary valve, the buffer solution II is connected to the inlet III through the rotary valve, and the buffer solution III is connected to the inlet IV through the rotary valve; the rotary valve is connected with the micro electromagnetic pump, and the micro electromagnetic pump provides power for the buffer solution to drive;
a 30-mesh and 60-mesh plastic gauze is fixedly arranged at the inlet of the quantitative electromagnetic pump;
the quantitative electromagnetic pump and the trace electromagnetic pump are connected with an upper computer.
Furthermore, the device also comprises an excitation circuit module and a detection circuit module which are connected to the electrode layer of the microfluid chip and are connected to the impedance test module and the electrophoresis test module.
Furthermore, the micro-excitation optical module is connected with an incident optical fiber embedded in the side wall of the micro-channel in the extraction fluorescence test chip module of the micro-fluidic chip through an optical fiber, and the micro-spectrum module is connected with an emergent optical fiber embedded in the side wall of the micro-channel in the extraction fluorescence test chip module of the micro-fluidic chip through an optical fiber.
Further, the microfluidic chip is sequentially provided with a top cover plate layer, a microfluidic channel substrate insulating layer, a detection electrode layer and a substrate fixing layer from top to bottom;
the top cover plate layer is made of polymer plastics and comprises a sample injection storage tank on the left side of the chip, a filter membrane supporting structure and three injection ports on the right side of the chip; the filter membrane supporting structure is placed in the sample injection storage tank, and the filter membrane is placed on the filter membrane supporting structure;
the filter membrane is in an array structure with 10 mu m aperture.
Furthermore, the height of the microfluidic channel layer is more than 100 μm, the microfluidic channel layer is also made of polymer plastic and is an elastoplast, and the elastoplast forms micro channels which are communicated with each other through a set structural shape and are used for liquid to flow in;
the microfluidic channel layer is provided with an optical fiber with the diameter of 100 mu m inserted at a specific position of the side wall of the extraction fluorescence test chip module; the receiving fiber should be at a 90 ° right angle to the incident fiber;
the micro-fluidic channel substrate insulating layer and the micro-channel are made of the same material, so that bonding connection is facilitated.
Furthermore, the position of the on-chip micro electromagnetic valve of the substrate insulating layer is provided with a specific step boss;
the thickness of the base insulating layer is more than 50 μm and less than 200 μm;
the height of the step projection is the half channel height.
Furthermore, the detection electrode layer is buried below the base insulating layer and is tightly attached to the substrate fixing layer;
the detection electrode layer is made of Au of 50-100nm and Cr or Ti of 5-10 nm.
On the other hand, the invention also discloses a multi-parameter rapid detection method for the water body, and the multi-parameter rapid detection device for the water body based on the method comprises the following steps:
firstly, buffer solution lavage is carried out; starting an upper computer control program, closing the quantitative electromagnetic pump, opening the on-chip micro electromagnetic valve I and the on-chip micro electromagnetic valve II, switching the rotary valve to a position for communicating the buffer solution I and the inlet II, opening the micro electromagnetic pump, filling the buffer solution I, washing air sealed in the micro channel than the extrusion channel, and closing the micro electromagnetic pump;
secondly, rinsing; a quantitative electromagnetic pump is adopted to provide a power source, the power source can carry out quantitative continuous delivery of samples, and sample liquid in a sample bottle is quantitatively delivered to a microfluidic channel through a filter cake; at the moment, the on-chip micro electromagnetic valve I and the on-chip micro electromagnetic valve II are in a closed state; the flow meter begins to record the flow rate v1 of the sample injection, and the flow meter begins to record the flow rate v2 of the waste liquid flowing to the outlet; when V1 is V2, starting automatic timing, and the time t1 for stably rinsing the microchannel is t1 more than 1 minute;
thirdly, sample introduction; starting the on-chip micro electromagnetic valve I, controlling the sample solution to enter the dielectrophoresis module, and starting timing when the v2 is stable;
fourthly, separating bacteria in the sample by dielectrophoresis; when v2 is stable, synchronously starting an exciting dielectrophoresis part in the exciting circuit module, and inputting a stable low-voltage sine high-frequency signal, preferably a sine wave signal with a peak value of 5Vpp and a frequency of 1 MHz; at the moment, bacteria are trapped in the micro-channel of the dielectrophoresis module due to dielectrophoresis force, the quantitative electromagnetic pump is closed, the time from v2 stabilization to the closing of the quantitative electromagnetic pump is recorded as t2, the sample entering amount is calculated through the difference value of the flow meter, and the entering sample amount is (v1-v2) t 2;
fifthly, detecting the number of bacteria; starting the micropump, switching the rotary valve to a buffer solution I pump inlet II, starting the on-chip micro electromagnetic valve I, starting the impedance bacteria test chip module in the detection circuit, reducing a sinusoidal signal input by an excitation dielectrophoresis part in the excitation circuit module to 3Vpp at the moment, enabling the buffer solution to have a set flow rate, flushing the intercepted bacteria into the impedance spectrum detection module along the edge of the dielectrophoresis microelectrode, and detecting the bacteria flowing through the impedance microelectrode by using the impedance microelectrode to judge the number of the bacteria; after no signal of the impedance spectrum is detected, the next step is carried out;
sixthly, extracting organic matters for fluorescence detection; closing the first micro electromagnetic valve on the chip, opening the second micro electromagnetic valve, and closing the second micro electromagnetic valve after t3, wherein the amount of the entering sample is (v1-v2) t 3; starting the micro electromagnetic pump, switching the rotary valve to a second buffer solution pump, and pumping the buffer solution into a third inlet; starting the micro spectrum module as an optical detection module and starting the micro laser light-emitting module as an excitation light module; the optical detection module is connected with an optical fiber and is directly inserted into the side wall of the micro-channel; calculating the content of organic matters in the sample by detecting a light intensity signal passing through a micro-channel at a specific detection part of the optical fiber, thereby completing the detection of the organic matters;
seventhly, detecting inorganic ions; the micro electromagnetic valve I on the chip is still closed, the micro electromagnetic pump is started, and the rotary valve is switched to the buffer solution III to be pumped into the inlet IV; applying direct-current high voltage of 1kv in the electrophoresis module, starting a non-contact conductive excitation circuit part in the excitation circuit module, inputting a stable low-voltage sine low-frequency signal, and starting a non-contact conductive detection circuit in the detection circuit; and calculating the content of the inorganic ions in the sample by detecting the voltage signal passing through the non-contact electric conduction electrode, thereby completing the detection of the inorganic ions.
Further, the method for determining the stability of v2 in the third step is as follows: the frequency acquired by the second flowmeter is 20Hz, and when the average speed v2 ' of the next 20 sampling points and the average speed v2 of the last 20 sampling points meet the formula | v2 ' -v2|/v2 ' < 5%, it can be determined that v2 is stable.
According to the technical scheme, compared with the prior art, the device and the method for rapidly detecting the multiple parameters of the water body overcome the defect that portable equipment for accurately and rapidly detecting the multiple parameters of the water body on site is unavailable. The method can rapidly and accurately measure the multiple parameters of the water body in the field, remarkably improves the test efficiency, remarkably reduces the test cost, and has the characteristics of good timeliness, simplicity and convenience in operation, and capability of accurately and rapidly obtaining the multiple parameter content of the water body.
The device adopts a micro-fluidic technology, realizes the accurate flow direction control of liquid through a micro-channel and a micro-valve, highly integrates a plurality of pretreatment and detection modules on a small plastic sheet, and has higher integration level and high-efficiency mass transfer and detection speed. The pretreatment of the water body is simple and only needs to be filtered. Therefore, the whole set of device has stronger portability and higher detection speed and timeliness. A plurality of modules are integrated on the same chip, so that multi-parameter joint detection is realized, and only one sample needs to be input, so that the detection accuracy is higher.
The electrochemical method has high detection sensitivity and extremely high detection response speed.
Drawings
FIG. 1 is a schematic structural view of the present invention;
FIG. 2 is a schematic diagram of a microfluidic chip structure and a schematic diagram of a fluid control flow direction according to the present invention;
FIG. 3 is a schematic view of the channel layer and electrode layer of the dielectrophoretic bacteria sorting chip module of the present invention;
FIG. 4 is an extracted fluorescence test module channel layer of the present invention;
FIG. 5 illustrates a channel layer and an electrode layer of the impedance testing module of the present invention;
FIG. 6 shows a channel layer and an electrode layer of an electrophoretic ion test module according to the present invention;
FIG. 7 is a schematic cross-sectional view of a microfluidic chip of the present invention in a resting state;
FIG. 8 is a schematic view of the microchannel cover plate layer structure of the present invention;
FIG. 9 is a schematic diagram of the structure and arrangement of the electrode layers of the dielectrophoretic bacteria sorting chip module of the invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are some, but not all, embodiments of the present invention.
As shown in fig. 1, the device for rapidly detecting multiple parameters of a water body according to the embodiment includes: the device comprises a microfluidic chip, an upper computer, a control circuit, a power supply module, a collection card, an excitation circuit module, a detection circuit module, a micro spectrum module, a micro laser light-emitting module, a quantitative electromagnetic pump, a micro electromagnetic pump, a rotary valve, a buffer bottle and a waste liquid bottle, and the like, wherein all the parts are connected as shown in figure 1.
The pipe of the sample bottle is connected to the inlet of the quantitative electromagnetic pump, 30-mesh and 60-mesh plastic gauze is fixedly placed in front of the inlet, and the outlet of the quantitative electromagnetic pump is connected with the inlet of the flowmeter 1. The outlet of the flowmeter 1 is connected with the inlet 1 of the micro-fluidic chip, the outlet of the micro-fluidic chip is connected with the inlet of the flowmeter 2, and the outlet of the flowmeter 2 is connected with the waste liquid. Buffer 1 is connected to inlet 2 by a rotary valve, buffer 2 is connected to inlet 3 by a rotary valve, and buffer 4 is connected to inlet 4 by a rotary valve. The micro electromagnetic pump provides power drive for the buffer solution. The control circuit controls the on-off of all pump valve circuits and the like, and the upper computer controls the whole system. The excitation circuit module and the detection circuit module are connected to an electrode layer of the microfluid chip and are connected to the impedance testing module and the electrophoresis testing module. The micro excitation optical module is connected with an incident optical fiber module embedded in the side wall of the micro channel in an extraction fluorescence test chip module of the micro-fluidic chip through an optical fiber, and the micro spectrum module is connected with an emergent optical fiber module embedded in the side wall of the micro channel in the extraction fluorescence test chip module of the micro-fluidic chip through an optical fiber.
The microchip for portable water quality multi-parameter quick test comprises a sample inlet, a waste liquid port, three buffer liquid ports and two micro electromagnetic valves on a chip, wherein the liquid flow direction of a micro channel on the control chip is a specific direction through the ports and the valves, as shown in figure 2, the situation of liquid return or reverse flow is prevented from occurring, and the test accuracy is not reduced.
The chip comprises four main modules, namely a dielectrophoresis bacteria sorting chip module, an impedance bacteria testing chip module, an extraction fluorescence testing chip module and an electrophoresis ion chip module.
The dielectrophoresis bacteria sorting chip module is positioned at the front end of the whole chip, the sample inlet channel is directly connected to the front end of the dielectrophoresis bacteria sorting electrode cavity, and the buffer solution inlet channel 1 is connected to the front end of the electrode cavity and converged to enter the front end of the dielectrophoresis bacteria sorting electrode cavity. The outside of the cavity is mainly a microchannel, the inside of the cavity is provided with a multilayer structure which comprises an electrode layer, the structure of the electrode layer is positioned in figure 9, the electrode layer is characterized in that convex rectangular, semicircular, triangular and trapezoidal microstructures are taken as main parts, and the arrangement mode is opposite and staggered. The outlet of the cavity is divided into two paths, one path is directly connected to the sample waste liquid outlet, and the other path is the liquid outlet of the dielectrophoresis module and is also directly connected to the liquid inlet of the impedance module. The principle is as follows: the bacteria are influenced by two forces of fluid force and dielectrophoresis force in the micro-channel, the fluid force is mainly controlled by flow rate, the dielectrophoresis force is determined by the convex structure and an applied alternating current signal, and the bacteria move towards the path of the inlet of the impedance module under the influence of the two forces after entering the sorting cavity.
The electrophoresis chip and the impedance bacteria test chip module also comprise an electrode layer. The electrode structure is shown in the figure. The excitation electrode and the receiving electrode are respectively connected to the excitation circuit module and the detection circuit module.
The extraction fluorescence test chip module has three ports in total, wherein the left side of the extraction fluorescence test chip module is provided with a fluorescence test module inlet, the right side of the extraction fluorescence test chip module is provided with a buffer solution inlet 2, and the liquid outlet 2. The entrance is divided into two paths after entering, a bend with 90 degrees is formed, and the exit optical fiber and the incident optical fiber are vertically arranged on the channel wall at the bend. The velocity of flow of sample is less than the buffer solution velocity of flow, and the left side sets up the flow direction of crescent and changes a way auxiliary structure, and the combined action makes the liquid flow direction turn to. Two pairs of optical fibers respectively test the fluorescence intensity of the sample inlet and the fluorescence intensity after the buffer solution extraction.
The micro-fluidic chip and the auxiliary clamp have five structures, namely a top cover plate layer, a micro-fluidic channel substrate insulating layer, a detection electrode layer and a substrate fixing layer from top to bottom in sequence.
The top cover plate layer is made of polymer plastic, and comprises a sample injection storage groove at the left side of the chip, a filter membrane supporting structure and three injection ports at the right side of the chip, as shown in fig. 8. The filter membrane is of an array structure with 10 mu m pore size, can filter impurities with the size of more than 10 mu m, and can allow bacteria with the size of less than 10 mu m to pass through and enter the microchannel.
The height of the micro-channel is larger than 100 μm, the micro-flow channel layer is also made of polymer plastic and is an elastoplast, and the elastoplast forms micro-channels which are communicated with each other through a certain structural shape and are used for liquid to circulate in the micro-channels. This layer mounts an optical fiber inserted 100 μm in diameter at a specific location on the sidewall of the extraction fluorescence test chip module. To improve the signal-to-noise ratio of the spectrometer signal and reduce the effect of incident light on the results, the receiving fiber should be at a 90 ° right angle to the incident fiber.
The micro-fluidic channel substrate insulating layer and the micro-channel are made of the same material, so that bonding connection is facilitated. The insulating base layer has specific step protrusions at the positions of the on-chip micro solenoid valves. Because the material has certain elasticity, when the on-chip electromagnetic valve works, the electromagnetic valve extends out to be jacked at the raised position, and the raised position is jacked up to be jacked to the position of the channel cover plate layer to block the liquid in the channel. The thickness of the insulating layer should be greater than 50 μm and less than 200 μm. The height of the protrusion is the half channel height. The solenoid valve extension also only requires half the channel height. Therefore, the liquid can be ensured to smoothly circulate when the elevator is static, and the dead volume is not easily formed below the step; on the other hand, the deformation amount is not too large during operation, so that the service life is not too low.
The electrode layer is buried under the insulating layer and closely attached to the substrate fixing layer. The electrode layer is made of Au of 50-100nm and Cr or Ti of 5-10 nm. The electrodes are only present at specific locations.
The novel field rapid detection method provided by the embodiment of the invention aims at three indexes of water quality detection: bacterial count, inorganic ion concentration, and organic matter.
Different from the traditional national standard detection method, the method directly detects the sample, does not need to additionally add reagents such as a marker, a color developing agent and the like, and does not cause secondary pollution to the environment. The invention is different from the existing single-index detection method or instrument, the sample is divided into several parts and then is injected, the invention adopts a complete micro-fluidic chip, the injected sample of the new detection method is the same sample, the same sample is injected to detect three indexes, partial system errors are eliminated, and the detection result is more reliable and convincing. Water samples such as river water, lake water and the like collected in the field need to be subjected to coarse filtration by using a clean filter screen, only solid impurities are removed, and water samples such as underground water, domestic drinking water and the like do not need to be subjected to coarse filtration. After a sample to be detected is sampled and uniformly mixed according to a standard sampling method, the other end of a sample inlet pipe with one end connected to a sample port on the left side of the microfluidic chip is inserted into a sample bottle, and the sample port is shown in figure 1.
In the first step, buffer lavage is performed. Starting an upper computer control program, closing the quantitative electromagnetic pump, opening the miniature electromagnetic valves 1 and 2 on the chip, switching the rotary valve to the positions of communicating the buffer solution 1 and the inlet 2, opening the micro electromagnetic pump, filling the buffer solution 1, and washing the air sealed in the micro channel extrusion channel. And turning off the micro electromagnetic pump.
And secondly, rinsing. A quantitative electromagnetic pump is adopted to provide a power source, the power source can carry out quantitative continuous conveying of samples, and sample liquid in a sample bottle is conveyed to a microfluidic channel through a filter cake in a quantitative mode. At this time, the on-chip micro solenoid valve 1 and the on-chip micro solenoid valve 2 are in a closed state. The flow meter 1 starts to record the flow rate v1 of the sample injection, and the flow meter 2 starts to record the flow rate v2 of the outlet flow to the waste liquid. When V1 becomes V2, the automatic timer is started, and the time t1 for the microchannel to be stably rinsed is set. Preferably, t1 should be greater than 1 minute.
And step three, sample introduction. And starting the on-chip micro electromagnetic valve 1 to control the sample solution to enter the dielectrophoresis module, and starting timing when the v2 is stable. The decision strategy for v2 stabilization is: the frequency acquired by the flowmeter 2 is several hertz to several tens of hertz, preferably 20Hz, and when the average velocity v2 ' of the next 20 sampling points and the average velocity v2 of the last 20 sampling points satisfy the formula | v2 ' -v2|/v2 ' × 100% < 5%, it can be determined that v2 is stable.
Fourthly, separating bacteria in the sample by dielectrophoresis. When v2 is stable, synchronously, the exciting dielectrophoresis part in the excitation circuit module is started, and a stable low-voltage sinusoidal high-frequency signal is input, preferably a sinusoidal signal with a peak value of 5Vpp and a frequency of 1 MHz. At this time, the bacteria are trapped in the micro-channels of the dielectrophoresis module due to dielectrophoretic forces. And (3) closing the quantitative electromagnetic pump, and calculating the sample entering amount through the difference of the flow meter, wherein the time from v2 stabilization to the closing of the quantitative electromagnetic pump is recorded as t2, and the entering sample amount is (v1-v2) t 2.
Principle of bacteria entrapment by dielectrophoresis: according to the micro-channel with the special microelectrode pattern, as shown in fig. 3, black is an electrode pattern, and the change of the electric field intensity in the channel is realized by arranging the electrode bulges, so that bacteria in a sample are trapped at the edge of the black electrode under the influence of dielectrophoresis force.
And fifthly, detecting the bacterial quantity. And (2) starting the micropump, switching the rotary valve to a buffer solution 1 to pump into an inlet 2, starting the on-chip micro electromagnetic valve 1, starting the impedance bacteria test chip module in the detection circuit, reducing the sine signal input by the dielectrophoresis exciting part in the exciting circuit module to 3Vpp at the moment, enabling the buffer solution to have a certain flow rate, flushing the intercepted bacteria into the impedance spectrum detection module along the edge of the dielectrophoresis microelectrode, and detecting the bacteria flowing through the impedance microelectrode by using the impedance microelectrode to judge the number of the bacteria. And after no signal is detected in the impedance spectrum, the next step is carried out.
And sixthly, extracting organic matters for fluorescence detection. And (3) closing the chip micro electromagnetic valve 1, opening the micro electromagnetic valve 2, and closing the micro electromagnetic valve 2 after t3, wherein the amount of the entering sample is (v1-v2) t 3. The micro electromagnetic pump is started, and the rotary valve is switched to the buffer solution 2 to be pumped into the inlet 3. At the moment, the micro spectrum module is started to serve as an optical detection module, and the micro laser light-emitting module is started to serve as an excitation light module. The optical detection module is connected with an optical fiber and is directly inserted into the side wall of the micro-channel. The content of organic matters in the sample is calculated by detecting the light intensity signal of the micro-channel passing through the specific detection part of the optical fiber, thereby completing the detection of the organic matters.
And seventhly, detecting inorganic ions. The on-chip micro electromagnetic valve 1 is still closed, the micro electromagnetic pump is started, and the rotary valve is switched to the buffer solution 3 to be pumped into the inlet 4. Applying direct-current high voltage 1kv in the electrophoresis module, starting a non-contact conductive excitation circuit part in the excitation circuit module, inputting a stable low-voltage sine low-frequency signal, and starting a non-contact conductive detection circuit in the detection circuit. And calculating the content of the inorganic ions in the sample by detecting the voltage signal passing through the non-contact electric conduction electrode, thereby completing the detection of the inorganic ions.
In conclusion, the device disclosed by the invention adopts a micro-fluidic technology, realizes the accurate flow direction control of liquid through the micro-channel and the micro-valve, highly integrates a plurality of pretreatment and detection modules on a small plastic sheet, and has higher integration level and high-efficiency mass transfer and detection speed. The pretreatment of the water body is simple and only needs to be filtered. Therefore, the whole set of device has stronger portability and higher detection speed and timeliness. A plurality of modules are integrated on the same chip, so that multi-parameter joint detection is realized, and only one sample needs to be input, so that the detection accuracy is higher. The electrochemical method has high detection sensitivity and extremely high detection response speed.
The above examples are only intended to illustrate the technical solution of the present invention, but not to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some technical features may be equivalently replaced; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions of the embodiments of the present invention.

Claims (10)

1. The utility model provides a be used for water multi-parameter short-term test device, includes the micro-fluidic chip module, its characterized in that:
the micro-fluidic chip module comprises a dielectrophoresis bacteria sorting chip module, an impedance bacteria testing chip module, an extraction fluorescence testing chip module and an electrophoresis ion chip module;
the microfluidic chip module is provided with four inlets, wherein the inlet I is used as a sample inlet, and the inlet II, the inlet III and the inlet IV are respectively used as a buffer solution inlet I, a buffer solution inlet II and a buffer solution inlet III;
the dielectrophoresis bacteria sorting chip module is respectively communicated with the first inlet and the sample waste liquid outlet through the micro-channel;
the dielectrophoresis bacteria sorting chip module is also communicated with the second inlet through a micro-channel;
the dielectrophoresis bacteria sorting chip module is communicated with the impedance bacteria test chip module through a micro-channel, and an on-chip micro electromagnetic valve I is arranged between the dielectrophoresis bacteria sorting chip module and the impedance bacteria test chip module;
the dielectrophoresis bacteria sorting chip module is respectively communicated with the extraction fluorescence test chip module and the electrophoresis ion chip module through microchannels by an on-chip micro electromagnetic valve II;
the extraction fluorescence test chip module is also communicated with the inlet III through a micro-channel, and the electrophoresis ion chip module is communicated with the inlet IV through a micro-channel;
the impedance bacteria test chip module, the extraction fluorescence test chip module and the electrophoresis ion chip module are respectively communicated with the first liquid outlet, the second liquid outlet and the third liquid outlet in a one-to-one correspondence manner through microchannels;
the device further comprises an upper computer, and the first micro electromagnetic valve and the second micro electromagnetic valve are respectively connected with the upper computer.
2. The device for rapidly detecting the multiple parameters of the water body according to claim 1, wherein:
the pipe of the sample bottle is connected to the inlet of the quantitative electromagnetic pump, and the outlet of the quantitative electromagnetic pump is connected with the inlet of the first flowmeter;
an outlet of the first flowmeter is connected with a sample inlet of the micro-fluidic chip, a sample waste liquid outlet of the micro-fluidic chip is connected to an inlet of the second flowmeter, and an outlet of the second flowmeter is connected to a waste liquid storage place;
the buffer solution I is connected to the inlet II through the rotary valve, the buffer solution II is connected to the inlet III through the rotary valve, and the buffer solution III is connected to the inlet IV through the rotary valve; the rotary valve is connected with the micro electromagnetic pump, and the micro electromagnetic pump provides power for the buffer solution to drive;
a 30-mesh and 60-mesh plastic gauze is fixedly arranged at the inlet of the quantitative electromagnetic pump;
the quantitative electromagnetic pump and the trace electromagnetic pump are connected with an upper computer.
3. The device for rapidly detecting the multiple parameters of the water body according to claim 1, wherein:
the device also comprises an excitation circuit module and a detection circuit module which are connected to the electrode layer of the microfluid chip and are connected to the impedance test module and the electrophoresis test module.
4. The device for rapidly detecting the multiple parameters of the water body according to claim 1, wherein:
the micro-excitation optical module is connected with an incident optical fiber embedded in the side wall of the micro-channel in the extraction fluorescence test chip module of the micro-fluidic chip through an optical fiber, and the micro-spectrum module is connected with an emergent optical fiber embedded in the side wall of the micro-channel in the extraction fluorescence test chip module of the micro-fluidic chip through an optical fiber.
5. The device for rapidly detecting the multiple parameters of the water body according to claim 1, wherein: the micro-fluidic chip is sequentially provided with a top cover plate layer, a micro-fluidic channel substrate insulating layer, a detection electrode layer and a substrate fixing layer from top to bottom;
the top cover plate layer is made of polymer plastics and comprises a sample injection storage tank on the left side of the chip, a filter membrane supporting structure and three injection ports on the right side of the chip; the filter membrane supporting structure is placed in the sample injection storage tank, and the filter membrane is placed on the filter membrane supporting structure;
the filter membrane is in an array structure with 10 mu m aperture.
6. The device for rapidly detecting the multiple parameters of the water body according to claim 5, wherein:
the height of the microfluidic channel layer is more than 100 mu m, the microfluidic channel layer is also made of polymer plastic and is an elastoplast, and the elastoplast forms micro channels which are communicated with each other through a set structural shape and are used for liquid to flow in;
the microfluidic channel layer is provided with an optical fiber with the diameter of 100 mu m inserted at a specific position of the side wall of the extraction fluorescence test chip module; the receiving fiber should be at a 90 ° right angle to the incident fiber;
the micro-fluidic channel substrate insulating layer and the micro-channel are made of the same material, so that bonding connection is facilitated.
7. The device for rapidly detecting the multiple parameters of the water body according to claim 5, wherein:
the substrate insulating layer is provided with a specific step bulge at the position of the on-chip micro electromagnetic valve;
the thickness of the base insulating layer is more than 50 μm and less than 200 μm;
the height of the step projection is the half channel height.
8. The device for rapidly detecting the multiple parameters of the water body according to claim 5, wherein:
the detection electrode layer is buried below the base insulating layer and is tightly attached to the substrate fixing layer;
the detection electrode layer is made of Au of 50-100nm and Cr or Ti of 5-10 nm.
9. A multi-parameter rapid detection method for water bodies, which is based on the multi-parameter rapid detection device for water bodies of any one of claims 1 to 8, and is characterized in that: the method comprises the following steps:
firstly, buffer solution lavage is carried out; starting an upper computer control program, closing the quantitative electromagnetic pump, opening the on-chip micro electromagnetic valve I and the on-chip micro electromagnetic valve II, switching the rotary valve to a position for communicating the buffer solution I and the inlet II, opening the micro electromagnetic pump, filling the buffer solution I, washing air sealed in the micro channel than the extrusion channel, and closing the micro electromagnetic pump;
secondly, rinsing; a quantitative electromagnetic pump is adopted to provide a power source, the power source can carry out quantitative continuous delivery of samples, and sample liquid in a sample bottle is quantitatively delivered to a microfluidic channel through a filter cake; at the moment, the on-chip micro electromagnetic valve I and the on-chip micro electromagnetic valve II are in a closed state; the flow meter begins to record the flow rate v1 of the sample injection, and the flow meter begins to record the flow rate v2 of the waste liquid flowing to the outlet; when V1 is V2, starting automatic timing, and the time t1 for stably rinsing the microchannel is t1 more than 1 minute;
thirdly, sample introduction; starting the on-chip micro electromagnetic valve I, controlling the sample solution to enter the dielectrophoresis module, and starting timing when the v2 is stable;
fourthly, separating bacteria in the sample by dielectrophoresis; when v2 is stable, synchronously starting an exciting dielectrophoresis part in the exciting circuit module, and inputting a stable low-voltage sine high-frequency signal, preferably a sine wave signal with a peak value of 5Vpp and a frequency of 1 MHz; at the moment, bacteria are trapped in the micro-channel of the dielectrophoresis module due to dielectrophoresis force, the quantitative electromagnetic pump is closed, the time from v2 stabilization to the closing of the quantitative electromagnetic pump is recorded as t2, the sample entering amount is calculated through the difference value of the flow meter, and the entering sample amount is (v1-v2) t 2;
fifthly, detecting the number of bacteria; starting the micropump, switching the rotary valve to a buffer solution I pump inlet II, starting the on-chip micro electromagnetic valve I, starting the impedance bacteria test chip module in the detection circuit, reducing a sinusoidal signal input by an excitation dielectrophoresis part in the excitation circuit module to 3Vpp at the moment, enabling the buffer solution to have a set flow rate, flushing the intercepted bacteria into the impedance spectrum detection module along the edge of the dielectrophoresis microelectrode, and detecting the bacteria flowing through the impedance microelectrode by using the impedance microelectrode to judge the number of the bacteria; after no signal of the impedance spectrum is detected, the next step is carried out;
sixthly, extracting organic matters for fluorescence detection; closing the first micro electromagnetic valve on the chip, opening the second micro electromagnetic valve, and closing the second micro electromagnetic valve after t3, wherein the amount of the entering sample is (v1-v2) t 3; starting the micro electromagnetic pump, switching the rotary valve to a second buffer solution pump, and pumping the buffer solution into a third inlet; starting the micro spectrum module as an optical detection module and starting the micro laser light-emitting module as an excitation light module; the optical detection module is connected with an optical fiber and is directly inserted into the side wall of the micro-channel; calculating the content of organic matters in the sample by detecting a light intensity signal passing through a micro-channel at a specific detection part of the optical fiber, thereby completing the detection of the organic matters;
seventhly, detecting inorganic ions; the micro electromagnetic valve I on the chip is still closed, the micro electromagnetic pump is started, and the rotary valve is switched to the buffer solution III to be pumped into the inlet IV; applying direct-current high voltage of 1kv in the electrophoresis module, starting a non-contact conductive excitation circuit part in the excitation circuit module, inputting a stable low-voltage sine low-frequency signal, and starting a non-contact conductive detection circuit in the detection circuit; and calculating the content of the inorganic ions in the sample by detecting the voltage signal passing through the non-contact electric conduction electrode, thereby completing the detection of the inorganic ions.
10. The method for rapidly detecting the multiple parameters of the water body according to claim 9, wherein the method comprises the following steps: the method for judging the stability of v2 in the third step comprises the following steps: the frequency acquired by the second flowmeter is 20Hz, and when the average speed v2 ' of the next 20 sampling points and the average speed v2 of the last 20 sampling points meet the formula | v2 ' -v2|/v2 ' < 5%, it can be determined that v2 is stable.
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