CN115166173A - Paper microfluidic system based on multiple detection of smart phone app and detection method - Google Patents

Paper microfluidic system based on multiple detection of smart phone app and detection method Download PDF

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CN115166173A
CN115166173A CN202210505960.8A CN202210505960A CN115166173A CN 115166173 A CN115166173 A CN 115166173A CN 202210505960 A CN202210505960 A CN 202210505960A CN 115166173 A CN115166173 A CN 115166173A
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熊小路
韩俊峰
郭程旺
许诗琪
宋红
姚裕贵
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Yangtze River Delta Research Institute Of Beijing University Of Technology Jiaxing
Beijing Institute of Technology BIT
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Abstract

The invention discloses a paper micro-fluidic system and a detection method based on multiple detection of smart phone app.A detection program is arranged in a smart phone, a paper micro-fluidic chip is shot by a camera of the smart phone, data analysis is carried out on the paper micro-fluidic chip image shot by an image acquisition module to obtain HSV (hue, saturation and value) of each detection image, and then the ion type and the ion concentration contained in a solution to be detected and the PH value of the solution to be detected are detected according to data stored in a database, so that the detection efficiency and the accuracy can be improved; through the app reading of the mobile phone, errors of naked eyes and a longer comparison process are avoided, and the detection result is more accurate and faster; the use of the mobile phone support enables result collection and analysis operation to be simpler and more convenient; 5 detection objects can be simultaneously and efficiently analyzed by optimizing the proportion of the reaction solution; the microfluidic paper chip system is easy to operate, high in efficiency and high in repeatability.

Description

Paper microfluidic system based on multiple detection of smart phone app and detection method
Technical Field
The invention belongs to the field of micromachining technology and information, and particularly relates to a paper micro-fluidic system based on multiple detection of smart phone app and a detection method, which are applied to accurate, efficient and multiple detection of water.
Background
The paper microfluidic chip has gained wide attention in a plurality of application fields such as water body pollution detection, food safety monitoring, medical diagnosis and the like. In recent years, the application of color detection to paper microfluidic chips has been advanced, the method analyzes multiple color signals through specific color reaction, so as to realize the simultaneous detection of multiple different pollutants in water, but the analysis of color signals by naked eyes is not accurate, and another computer software analysis method based on color reaction images has no timeliness. Therefore, the development of an accurate and efficient multiple detection method is an urgent problem to be solved for the water quality detection application of the paper microfluidic chip.
The smart phone brings a new vitality for the application of the paper microfluidic chip color detection, and initially, a mobile phone camera becomes an important image acquisition tool, so that inaccurate judgment of human eyes on analysis signals is avoided, and the accuracy of analysis and comparison of color signals is effectively improved. At present, various breakthroughs are made in functions of smart phone application programs, and the smart phone application programs have the functions of image recognition, data analysis algorithms and the like. Therefore, the advantages of the application program of the smart phone are expected to be utilized, the functions of water quality data acquisition, storage and analysis are combined, and the real-time water quality monitoring system with high portability and strong functionality is developed and applied to high-efficiency and multiple water quality detection and analysis.
Disclosure of Invention
In view of the above, the invention provides a paper microfluidic system and a detection method based on multiple detections of smart phone app, the whole detection process only needs 5-10 minutes, metal ions, a pH value and nitrite can be distinguished and displayed through a mobile phone program, and the content of each component can be analyzed within a certain range.
A paper microfluidic system based on multiple detection of smart phone app comprises a smart phone and a paper microfluidic chip;
the flow channel pattern of the paper microfluidic chip comprises a central area (1) and a plurality of detection areas (2) distributed around the periphery of the central area (1); each detection area (2) is communicated with the central area (1) through a micro-channel; different detection solutions are respectively dripped into each detection area (2), and the detection solutions comprise a detection solution for detecting various heavy metal ions, a detection solution for detecting a PH value and a detection solution for detecting nitrite ions; the central area (1) is used for dripping the liquid to be detected;
the smart phone is provided with a detection program, and the detection program is provided with an image acquisition module, a database and an analysis detection module;
the image acquisition module is used for driving a camera of the smart phone to shoot the paper microfluidic chip;
after each detection area detects corresponding ions, a corresponding relation curve between HSV (hue, saturation, value) and ion logarithmic concentration of an image of the detection area and a relation curve between HSV and PH value of the image of the detection area are stored in the database;
the analysis detection module is used for carrying out data analysis on the paper microfluidic chip image shot by the image acquisition module to obtain HSV values of all detection images, and then detecting the ion species and the ion concentration contained in the solution to be detected and the PH value of the solution to be detected according to data stored in the database.
Preferably, the HSV value calculation method of the image is as follows:
R′＝R/255,G′＝G/255,B′＝B/255,
Cmax＝max(R′,G′,B′),Cmin＝min(R′,G′,B′),Δ＝Cmax-Cmin
r, G, B are the values of R, G, B three channels of the image respectively;
the values for the R, G, B triple channel are then converted to HSV space:
Figure BDA0003636162130000021
Figure BDA0003636162130000022
V＝Cmax。
preferably, the paper microfluidic chip is realized in a way of wax printing a flow channel pattern on filter paper.
Preferably, the central area (1) of the flow channel pattern is circular.
Preferably, the detection area (2) is shaped as a sector ring.
Preferably, the smart phone is further provided with a display module for displaying the detection result obtained by the analysis and detection module.
Further, the device also comprises a support which is divided into an upper layer and a lower layer, wherein the upper layer is used for storing the smart phone, and the lower layer is used for placing the paper micro-fluidic chip.
A detection method of a paper microfluidic system based on multiple detection comprises the steps of preparing a detection solution, wherein the detection solution comprises a copper ion detection agent, a nickel ion detection solvent, an iron ion detection agent, a nitrite ion detection solvent and a pH value detection reagent.
Preferably, the preparation method of the detection solution is as follows:
mixing hydroxylamine hydrochloride, ammonium acetate, bathocuproine-acetone solution and PEG 600 according to the proportion of 1;
mixing 0.5M sodium fluoride solution, 6.3M acetic acid solution, 20mg/ml dimethylglyoxime acetone and ammonia water solution according to the weight ratio of 2:1:5:2.5, mixing the components in proportion to prepare a nickel ion detection solvent;
330mM citric acid, 50mM sulfanilamide and 10mM NED, as 1:1:1, and mixing. Preparing a nitrite ion detection agent;
mixing ammonium acetate, 1, 10-phenanthroline and hydroxylamine hydrochloride according to the weight ratio of 1:2:1 to prepare an iron ion detection agent;
the bromothymol blue and ethanol are used as pH value detection reagents according to the proportion of 10%.
Preferably, the detection method further comprises:
(1) Running the application program and starting detection;
(2) Selecting a chip pattern in an application program according to an actually used paper micro-fluidic chip;
(3) Respectively adding 2.5 mul of copper ion detection agent, iron ion detection agent and nitrite detection agent with different concentrations into different detection areas (2); then adding 30 mul of solution to be detected into the central area (1), and slowly allowing the solution to be detected to enter each detection area (2) from the central area (1) along the flow channel after the solution is completely soaked;
(4) The image acquisition module captures an image, and a picture of the chip is displayed on a screen; a database and;
(5) The analysis and detection module is used for analyzing and calculating the image to obtain the ion species, the ion concentration and the pH value of the solution to be detected;
(6) And displaying the detection result on the screen of the mobile phone.
The invention has the following beneficial effects:
the invention provides a paper microfluidic system and a detection method based on multiple detections of smart phone app, wherein a detection program is arranged in a smart phone, a paper microfluidic chip is shot by a camera of the smart phone, data analysis is performed on an image of the paper microfluidic chip shot by an image acquisition module to obtain HSV (hue, saturation, value) values of all detected images, and then the types and the concentrations of ions contained in a solution to be detected and the PH value of the solution to be detected are detected according to data stored in a database, so that the detection efficiency and the detection accuracy can be improved;
through mobile phone app reading, errors of naked eyes and a longer comparison process are avoided, and a detection result is more accurate and faster;
the use of the mobile phone support enables result collection and analysis operation to be simpler and more convenient;
5 detection objects can be simultaneously and efficiently analyzed by optimizing the proportion of the reaction solution;
the microfluidic paper chip system is easy to operate, high in efficiency and high in repeatability.
Drawings
FIG. 1 is a schematic diagram of a paper microfluidic chip;
FIG. 2 is a schematic structural view of a handset and a paper chip holder;
fig. 3 is a diagram of 4 interfaces of the smartphone.
Detailed Description
The invention is described in detail below by way of example with reference to the accompanying drawings.
The invention researches and optimizes the shape, the size, the baking time, the solution components of the reaction liquid and other experimental parameters of the wax printing paper microfluidic chip, develops an application program of the smart phone, and can record and simultaneously detect the chrominance signals of 5 analytes in real time. The paper microfluidic system based on the multiple detection of the smartphone app reading improves the analysis efficiency of color detection and the convenience of reading results by a user, and has great potential in water quality data collection and analysis decision.
The paper microfluidic system comprises a smart phone, a paper microfluidic chip and a bracket; as shown in fig. 1, the paper microfluidic chip is implemented by wax printing a channel pattern on filter paper, wherein the channel pattern comprises a circular central area 1, and 6 fan-ring-shaped detection areas 2 are distributed around the periphery of the central area 1; each detection zone 2 is in communication with the central zone 1 via a microchannel. The other areas except the flow channel patterns on the filter paper are soaked by the wax liquid and are hydrophobic areas, and the flow channel patterns are not soaked by the wax liquid and are hydrophilic areas. Because the main component cellulose of the filter paper enables the solution to flow through capillary action and is a hydrophilic area, and an external driving pump is not needed, the solution to be detected can flow into the surrounding detection area 2 after being dripped into the central area 1; the areas outside the flow channel pattern are hydrophobic areas formed by the wax liquid, and the solution cannot infiltrate and flow, so the solution flows only along the designed flow channel areas.
Different detection solutions are respectively dropped into each detection area 2, and the detection solutions comprise a detection solution for detecting various heavy metal ions, a detection solution for detecting a PH value and a detection solution for detecting nitrite ions; the central area 1 is used for dripping the liquid to be detected.
The smart phone is provided with a detection program, and the detection program is provided with an image acquisition module, a database, an analysis detection module and a display module.
The image acquisition module is used for driving a camera of the smart phone to shoot the paper micro-fluidic chip.
After the corresponding ions are detected in each detection area, a corresponding relation curve between the HSV value and the ion logarithmic concentration of the image in the detection area and a relation curve between the HSV value and the PH value of the image in the detection area are stored in the database.
The HSV value calculation method of the image comprises the following steps:
R′＝R/255,G′＝G/255,B′＝B/255,
Cmax＝max(R′,G′,B′),Cmin＝min(R′,G′,B′),Δ＝Cmax-Cmin (1)
r, G, B is the value of R, G, B three channels of the image respectively;
the values of R, G, B triple channel are then converted to HSV space:
Figure BDA0003636162130000061
the analysis detection module is used for carrying out data analysis on the paper microfluidic chip image shot by the image acquisition module, obtaining HSV values of all detection images according to a formula (1) and a formula (2), then detecting the ion type and the ion concentration contained in the solution to be detected and the PH value of the solution to be detected according to data stored in a database, and displaying the information through the display module.
In this embodiment, the linear relationship between the logarithmic concentrations of Cu (II), ni (II), fe (iii), and nitrides, and the pH of the solution and the HSV value of the image is as follows:
Cu(II)：y＝46.0x-17.6(R 2 ＝0.978)；
Ni(II)：y＝31.1x-9.41(R 2 ＝0.960)；
Fe(Ⅲ)：y＝57.8x-103(R 2 ＝0.986)；
nitrite salt: y =47.6x-7.83 (R) 2 ＝0.960)；
pH value: y =24.7x-22.2 (R) 2 ＝0.982)。
x represents logC (logarithmic value of concentration C), y represents gray value =255- (R + G + B)/3,R 2 Representing the correlation of the fitted linear equation. The closer to 1, the higher the degree of anastomosis.
As shown in fig. 2, the bracket for fixing the smart phone and the paper microfluidic chip is divided into an upper layer and a lower layer, wherein the upper layer is used for storing the smart phone, and the lower layer is used for placing the paper microfluidic chip. The camera of the smart phone is aligned with the paper micro-fluidic chip and can be directly photographed.
The manufacturing method of the paper microfluidic chip comprises the following steps:
firstly, using adobe illustrator software to design a flow channel pattern of the paper microfluidic chip; then, manufacturing a wax printing chip, which specifically comprises the following steps:
installing a wax block in a wax box area, putting whatman 1 filter paper in a paper feeding area of a printer, and setting a printing mode to be color printing to obtain a wax printing pattern; placing wax printing filter paper on a hot plate, and heating at 150 ℃ for 5 minutes;
preferably, before the paper microfluidic chip is used, the back surface of the chip is adhered with a transparent adhesive tape to prevent the solution from leaking.
Preferably, the diameter of the central area (1) is 6mm, the side length of the inner side of the outermost detection area (2) is 7.6mm, and the side length of the inner side of the outermost detection area is 6mm; the length of the micro flow channel between the central zone (1) and the detection zone (2) is designed to be 2.4mm. If the flow channel is long, the solution needs to travel a longer distance to reach the detection area, which may result in non-uniform color intensity in the detection area.
Preferably, the detection area is specially designed to be in a sector ring shape, the outer side of the detection area is in an arc shape, and the shape considers the flowing direction of the solution in the flow channel, so that the solution can be promoted to be uniformly distributed in the detection area, and the detection area is better than the common circular shape.
Preferably, in the second step, the baking temperature is set to 150 ℃ and the baking time is set to 5 minutes. If the baking time is too short, the wax does not completely penetrate the entire paper chip, especially the back side of the paper, and the solution in the flow path leaks.
The preparation method of the detection solution comprises the following steps:
the detection reaction solution for each analyte comprises a reducing agent, a color developing agent, a buffer solution, a masking agent and the like which are prepared into a mixed detection solution according to a certain proportion, wherein the mixed detection solution comprises a copper ion detection agent, a nickel ion detection solvent, an iron ion detection agent, a nitrite ion detection solvent and a detection reagent for pH value.
The method comprises the following specific steps: the copper ion detection agent is prepared by mixing hydroxylamine hydrochloride, ammonium acetate, bathocuproine-acetone solution (50 mg/ml), PEG 600 according to the proportion of 1. 0.5M sodium fluoride solution, 6.3M acetic acid solution, 20mg/ml dimethylglyoxime acetone and ammonia solution according to the weight ratio of 2:1:5:2.5 to prepare the nickel ion detection solvent. Nitrite ions were detected using the Griess method. Griess reagent included 330mM citric acid, 50mM sulfanilamide, and 10mM NED, as 1:1:1, and mixing. Mixing ammonium acetate, 1, 10-phenanthroline and hydroxylamine hydrochloride according to the weight ratio of 1:2:1 to prepare the iron ion detection agent. The bromothymol blue and ethanol are used as pH value detection reagents according to the proportion of 10%. In addition, solutions of nitride, pH, cu (II), ni (II), and Fe (III) were prepared as standard solutions.
The paper micro-fluidic system is used for detecting pH, nitride, copper ions, nickel ions and nitrite, and comprises the following specific steps:
(1) Running the application and selecting the "start" button to start the test;
(2) And selecting a chip pattern in an application program according to the actually used paper microfluidic chip.
(3) Adding 2.5 mul of copper ion detection agent, iron ion detection agent and nitrite detection agent with different concentrations into different detection areas 2 respectively. Then 30. Mu.l of the solution to be assayed are addedThe central area 1 is completely soaked by the solution, the solution to be detected slowly enters the detection area 2 from the central area 1 along the flow channel, and in the copper ion detection area 2, the detection area 2 is changed from colorless to orange, the orange color is different in shade, and the concentration of the solution is different. The detection principle is as follows: bathocuproing as color developing agent and Cu 2+ The reaction generates a dark yellow compound, and the Griess reagent is used as a color developing agent to react with nitrite to produce a purple red compound. Use of dimethylglyoxime as a color developer and Ni 2+ The reaction produced a pink complex. Bromothymol blue increases with basicity to form an orange or green complex, and phenanthroline reacts with Fe (III) to form an orange complex. The paper microfluidic chip can be used for simultaneously detecting pH, nitride, copper ions, nickel ions and nitrite, and can also be used for detecting other water quality components.
(4) Click the "new test" button to capture an image of the test result, and display a picture of the chip on the screen.
(5) And selecting an ANSYS button, running calculation according to an algorithm of an application program, and analyzing to obtain the ion name, the concentration and the PH value.
(6) And displaying the detection result. The ionic names (Cu (II), ni (II), fe (III), NO) contained in the liquid to be detected 2 And pH) and the corresponding concentration and pH value of the solution are displayed on the screen of the application.
In summary, the above description is only a preferred embodiment of the present invention, and is not intended to limit the scope of the present invention. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims (10)

1. A paper microfluidic system based on multiple detection of smart phone app is characterized by comprising a smart phone and a paper microfluidic chip;
the flow channel pattern of the paper microfluidic chip comprises a central area (1) and a plurality of detection areas (2) distributed around the periphery of the central area (1); each detection area (2) is communicated with the central area (1) through a micro-channel; different detection solutions are respectively dripped into each detection area (2), and the detection solutions comprise a detection solution for detecting various heavy metal ions, a detection solution for detecting a PH value and a detection solution for detecting nitrite ions; the central area (1) is used for dripping the liquid to be detected;
the smart phone is provided with a detection program, and the detection program is provided with an image acquisition module, a database and an analysis detection module;
the image acquisition module is used for driving a camera of the smart phone to shoot the paper microfluidic chip;
after the corresponding ions are detected in each detection area, a corresponding relation curve between the HSV value and the ion logarithmic concentration of an image of the detection area and a relation curve between the HSV value and the PH value of the image of the detection area are stored in the database;
the analysis detection module is used for carrying out data analysis on the paper microfluidic chip image shot by the image acquisition module to obtain HSV values of all detection images, and then detecting the ion species and the ion concentration contained in the solution to be detected and the PH value of the solution to be detected according to data stored in the database.
2. The smart phone app based multi-detection paper microfluidic system of claim 1, wherein HSV value of an image is calculated as follows:
R′＝R/255,G′＝G/255,B′＝B/255,
C max＝max(R′,G′,B′),C min＝min(R′,G′,B′),Δ＝C max-C min
r, G, B is the value of R, G, B three channels of the image respectively;
the values of R, G, B triple channel are then converted to HSV space:
Figure FDA0003636162120000011
Figure FDA0003636162120000012
V＝Cmax。
3. the smartphone app based multiple-detection paper microfluidic system of claim 1, wherein the paper microfluidic chip is implemented in the form of wax printing a channel pattern on filter paper.
4. The smartphone app based multi-detection paper microfluidic system of claim 1 or 2, wherein a center region (1) of a flow channel pattern is circular.
5. A smartphone app based multi-detection paper microfluidic system as claimed in claim 4, characterised in that the detection zone (2) is in the shape of a fan ring.
6. The smart phone app based paper microfluidic system according to claim 1 or 2, wherein a display module is further disposed in the smart phone for displaying the detection result obtained by the analysis detection module.
7. The smart phone app based multiple detection paper microfluidic system according to claim 1 or 2, further comprising a support divided into an upper layer and a lower layer, wherein the upper layer is used for storing a smart phone, and the lower layer is used for placing a paper microfluidic chip.
8. The detection method of the paper microfluidic system based on the multiplex detection of claim 1, comprising preparing a detection solution, wherein the detection solution comprises a copper ion detection agent, a nickel ion detection solvent, an iron ion detection agent, a nitrite ion detection solvent and a pH value detection reagent.
9. The detection method according to claim 8, wherein the detection solution is prepared by:
mixing hydroxylamine hydrochloride, ammonium acetate, bathocuproine-acetone solution and PEG 600 according to the proportion of 1;
mixing 0.5M sodium fluoride solution, 6.3M acetic acid solution, 20mg/ml dimethylglyoxime acetone and ammonia water solution according to the weight ratio of 2:1:5:2.5, mixing the components in proportion to prepare a nickel ion detection solvent;
330mM citric acid, 50mM sulfanilamide and 10mM NED, as 1:1:1, and mixing. Preparing a nitrite ion detection agent;
mixing ammonium acetate, 1, 10-phenanthroline and hydroxylamine hydrochloride according to the weight ratio of 1:2:1 to prepare an iron ion detection agent;
the bromothymol blue and ethanol are used as pH value detection reagents according to the proportion of 10%.
10. The detection method according to claim 8 or 9, characterized in that the detection method further comprises:
(1) Running the application program and starting detection;
(2) Selecting a chip pattern in an application program according to an actually used paper micro-fluidic chip;
(3) Respectively adding 2.5 mul of copper ion detection agent, iron ion detection agent and nitrite detection agent with different concentrations into different detection areas (2); then adding 30 mul of solution to be detected into the central area (1), and slowly allowing the solution to be detected to enter each detection area (2) from the central area (1) along the flow channel after the solution is completely soaked;
(4) The image acquisition module captures an image, and a picture of the chip is displayed on a screen; a database and;
(5) The analysis and detection module is used for analyzing and calculating the image to obtain the ion species, the ion concentration and the pH value of the solution to be detected;
(6) And displaying the detection result on a mobile phone screen.
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