CN115267046A - Dynamic light-regulated biological colorimetric detection method and system combined with mobile phone and tablet computer - Google Patents

Abstract

The invention relates to a clinical rapid diagnosis and instant detection technology, and aims to provide a dynamic light-regulated biological colorimetric detection method and system combined by a mobile phone and a tablet personal computer. The method is characterized in that an intelligent tablet computer is used as a bottom light source of a microtiter plate in a completely black environment, and a smart phone is used as a control terminal device to realize light source regulation and control, obtain contrast color images, and perform data processing analysis and result display. Compared with the traditional board reader, the invention utilizes the display screen of the intelligent tablet computer as a single light source, and can greatly expand the detection range of the system by changing the light source intensity and analyzing different color channels. The time of one experiment is less than 2 seconds, so that the detection time is greatly shortened, and the detection efficiency is improved; the color of the light source can be converted at any time according to different detection objects so as to obtain higher detection performance and improve the detection sensitivity; can be adapted to microtiter plates of various specifications of commercial products, thereby further reducing the use cost of users and having great application prospect.

Description

Dynamic light regulation biological colorimetric detection method and system combined with mobile phone and tablet computer

technical field

The invention relates to clinical rapid diagnosis and real-time detection technology, in particular to a dynamic light regulation biological colorimetric detection method and system used in conjunction with a mobile phone and a tablet computer.

Background technique

In the fields of clinical diagnosis, environmental monitoring and food safety testing, there is a great demand for simple, efficient and cheap detection technologies. The ability to perform at-home, simple, and high-speed point-of-care tests has attracted considerable attention in both academia and industry. The user-friendly nature of point-of-care testing allows non-professionals to implement biochemical testing in resource-limited settings, making it suitable for mass population screening during public events. Therefore, point-of-care tests are playing an increasingly important role in biochemical testing and have undergone rapid development in the past few decades. Many analytical methods, such as colorimetric assays and electrochemical measurements, are thought to improve the accuracy and sensitivity of point-of-care assays.

Because of its simple operation, fast response, strong adaptability and wide linear range, colorimetry is a point-of-care detection technique that can be widely used. Several colorimetric methods have also been developed to detect various biochemical samples, such as heavy metals in water, biomarkers in blood or environmental pollutants in sweat and air, by visual inspection or using microtiter plate readers. However, visual inspection can only qualitatively detect color changes in colorimetric assays, but cannot quantify analyte concentrations in biochemical samples. Meanwhile, expensive microtiter plate readers, although capable of determining the concentration of biochemical samples in a high-throughput manner, have limited application in point-of-care colorimetric assays due to cost constraints.

Therefore, it is imperative to develop portable and cost-effective analytical instruments for quantitative testing by colorimetric analysis in a high-throughput manner to achieve accurate and immediate detection based on colorimetric analysis.

Contents of the invention

The technical problem to be solved by the present invention is to overcome the deficiencies in the prior art, and provide a dynamic light-regulated biological colorimetric detection method and system used in combination with a mobile phone and a tablet computer.

For solving technical problem, solution of the present invention is:

Provides a dynamic light control biological colorimetric detection method combined with a mobile phone and a tablet computer. In a completely dark environment, the smart tablet computer is used as the bottom light source of the microtiter plate, and the smart phone is used as the control terminal device to realize the control of the light source and obtain contrasting color images. , data processing analysis and result display;

Described detection method specifically comprises the following steps:

(1) The interaction between the smart phone and the smart tablet is realized through wireless communication, and the smart phone sends light control instructions to the smart tablet according to the type of microtiter plate and the type of the substance to be detected to control the range, color and color of the latter’s light. Brightness, forming a light source dot matrix corresponding to each well on the microtiter plate;

(2) After the smart tablet computer completes the display of the specified light source, it sends a ready message to the smart phone, and the smart phone automatically takes pictures of the microtiter plate to obtain a colorimetric image;

(3) Deriving the RGB and HSL channels of each hole from the acquired colorimetric image, using the adaptive threshold algorithm to convert the colorimetric image into a binary image; then determine the positions of all circles in the image through Hough circle transformation, and obtain The center information of each hole; calculate the average pixel intensity of the 11×11 center pixel array of each hole, compare the calculation result with the standard marker curve of the detected object in the database, obtain the detection result of each hole and display it on the smartphone display.

As a preferred solution of the present invention, the method further includes step (4): the smart phone automatically uploads the detection results to the cloud server, and the cloud server performs data sorting and analysis according to preset rules.

As a preferred solution of the present invention, the type selection of the microtiter plate is manually input through the operation interface of the smart phone, or is automatically recognized after being photographed by the smart phone; the type of the detected substance is manually input through the operation interface of the smart phone enter.

As a preferred solution of the present invention, the microtiter plate includes several holes arranged in an array, each hole has a transparent bottom plate and a black upper structure, and the connecting parts between each hole are also black; The dot matrix of the light source corresponds to the transparent bottom plate of each well on the microtiter plate.

As a preferred solution of the present invention, the database is set on a cloud server, or built into a local memory of a smart phone.

The present invention further provides a dynamic light control biological colorimetric detection system used in conjunction with a mobile phone and a tablet computer, which includes a fully enclosed and light-tight box structure; a smart tablet with a display screen upward is fixed on the base surface of the box The computer and the microtiter plate are suspended above the smart tablet by the bracket and kept at an appropriate distance; there is a through hole on the top of the box, and the smart phone is placed on the top of the box with the display screen upward, and can use the through hole and the rear camera The microtiter plate is photographed; the smart phone has a built-in image detection and processing analysis module, which is used to obtain colorimetric images, data processing and analysis and result display according to the aforementioned method.

As a preferred solution of the present invention, the system also includes a cloud server capable of data interaction with the smart phone for storing calibration curves of different substances and/or analyzing and organizing data uploaded by the smart phone.

As a preferred solution of the present invention, the bracket is a drawable guide rail bracket, including four bearings arranged on the box body, every two bearings form a group and are connected with guide rails respectively, and the two guide rails are arranged in parallel; On the guide rail, the microtiter plate is embedded in the mounting position of the bracket.

As a preferred scheme of the present invention, the aperture of the through hole on the top of the box is 3cm, and the distance between the rear camera of the smart phone and the microtiter plate is 30cm.

Compared with prior art, the beneficial effect of the present invention is:

1. Compared with the traditional plate reader, the present invention uses the display screen of the smart tablet computer as a single light source, and can greatly expand the detection range of the system by changing the intensity of the light source and analyzing different color channels.

2. After the traditional plate reader acquires the colorimetric image, it still needs to manually count the data and draw the calibration curve. The detection time of one experiment exceeds 200 seconds. The invention automatically processes the colorimetric image and automatically draws the calibration curve, and the time for one experiment is less than 2 seconds, which greatly shortens the detection time and improves the detection efficiency.

3. The present invention can also flexibly switch the light source according to the needs of the detection substances, and change the color of the light source at any time according to different detection objects to obtain higher detection performance and improve detection sensitivity. Therefore, the present invention can be effectively used in various colorimetric applications.

4. The present invention uses the display screen of the smart tablet computer as the bottom light source of the microtiter plate, and can select a light source lattice with a corresponding shape and layout according to different types of microtiter plates, and it is convenient and almost cost-free to replace the light source lattice. Therefore, the present invention can adapt to commercially available microtiter plates of various specifications, thereby further reducing the user's use cost.

5. Compared with the general price of thousands of dollars to tens of thousands of dollars for scientific instruments such as traditional plate readers, smartphones and tablets are high-performance mobile devices with relatively low cost, large quantities, and wide coverage, which can provide a powerful platform Hardware support. These devices contain important modules such as high-resolution cameras, computer processors, and multiple sensors, and are small enough to be portable instruments, effectively keeping the manufacturing cost in the hundreds of dollars, so they are very popular. prospect.

Description of drawings

Figure 1 is a schematic diagram of the device structure.

Reference signs in the figure: 1 box body; 2 smart tablet computer; 3 micro titer plate; 4 bracket; 5 guide rail; 6 bearing; 7 smart phone; 8 base.

Figure 2 is the display interface (b) of the black well wall microtiter plate (a) and the smart tablet computer.

Figure 3 is the main interface of the smart phone APP as an example.

Figure 4 is an example of the smartphone APP colorimetric image acquisition interface.

FIG. 5 is a display diagram of smart phone APP data as an example.

Fig. 6 is the final result display interface of the smart phone APP as an example.

Figure 7 is the colorimetric imaging of tartrazine solution under different blue light source intensities.

Fig. 8 is the colorimetric image quantitative analysis of tartrazine solution.

In the figure: (a) the average pixel intensity of the blue channel with different light intensities under the blue light source; (b) the average pixel intensity of the green channel with different light intensities under the blue light source; (c) the average pixel intensity of the blue channel with different light intensities under the blue light source Detection range; (d) The detection range of the green channel with different light intensities under the blue light source.

Figure 9 is a comparison of the detection performance of the biological colorimetric detection system and the traditional plate reader;

In the figure: (a) is the best calibration curve, (b) is the corresponding detection range.

Figure 10 is a comparison of the detection time between the biological colorimetric detection system and the traditional plate reader.

Figure 11 shows the detection performance of amaranth solution under green light source and red light source.

In the figure: (a) colorimetric imaging of amaranth solution under green light source; (b) quantitative analysis of amaranth solution under green channel of green light source; (c) colorimetric analysis of amaranth solution under red light source Imaging; (d) Quantitative analysis of amaranth solution under the red channel of red light source;

Figure 12 is the detection performance of the biological colorimetric detection system and the traditional plate reader to the amaranth solution of the same concentration gradient;

Figure 13 is the application of the biological colorimetric detection system to the detection of phenol red solution;

In the figure: (a-b) phenol red colorimetric images recorded by the biological colorimetric detection system under the irradiation of blue light source and green light source; (c-d) blue channel calibration curve and detection concentration of phenol red solution obtained under different blue light intensities Range; (e-f) blue channel calibration curve and detection range of phenol red solution concentration obtained under different green light intensities; (g-h) best calibration curve obtained under blue light source and green light source and corresponding detection range of phenol red .

Figure 14 shows the application of the biological colorimetric detection system to the detection of cell number.

In the figure: (a) CCK-8 cell viability bioassay images recorded by the biological colorimetric detection system under green light irradiation; (b-e) green channel calibration curves of CCK-8 cell viability bioassays obtained under different light intensities of the green light source , detection range, goodness-of-fit (R2) value and detection sensitivity.

Figure 15 shows the application of the biological colorimetric detection system to the detection of BSA.

In the figure: (a) BSA image recorded by the biological colorimetric detection system under blue light source; (b-c) blue channel calibration curve and BSA detection range obtained under different light intensities of green light source.

Detailed ways

First of all, it needs to be explained that the present invention relates to image processing and database technology, and is an application of computer technology in the field of analysis and testing technology. During the implementation of the present invention, the application of multiple software function modules will be involved. The applicant believes that, after carefully reading the application documents and accurately understanding the realization principle and purpose of the present invention, combined with existing known technologies, those skilled in the art can fully implement the present invention by using their software programming skills. The aforementioned software functional modules include but are not limited to: image detection and processing analysis modules, etc., all mentioned in the application documents of the present invention belong to this category, and the applicant will not list them one by one.

The microtiter plate of the present invention can choose any style of commercially available commodity. Microtiter plates consist of wells (wells) with clear bottoms and solid polystyrene black upper structures suitable for fluorescent and luminescent biocolorimetric detection for cell culture and microscopy applications.

As shown in Fig. 1, the dynamic light control biological colorimetric detection system used in combination with mobile phones and tablet computers in the present invention includes a fully enclosed and light-tight box structure; a display is fixed on the surface of the base 8 of the box 1. The smart tablet computer 2 with the screen upwards, the microtiter plate 3 is suspended above the smart tablet computer 2 by the bracket 4 and kept at an appropriate distance; a through hole is provided on the top of the box, and the smart phone 7 is placed on the top of the box with the display screen facing upwards. And the through hole and the rear camera can be used to shoot the microtiter plate 3; as an optional solution, the support 4 is a drawable guide rail support, including four bearings 6 arranged on the casing 1, and every two bearings 6 are a group and are connected by guide rails 5 respectively, and the two guide rails 5 are arranged in parallel; As an example, the aperture of the through hole on the top of the box is 3 cm, and the distance between the rear camera on the smart phone 7 and the microtiter plate 3 is 30 cm.

The image detection and processing analysis module is built in the smart phone 3, which is used to realize the acquisition of colorimetric images, data processing and analysis, and result display. The system also includes a cloud server capable of data interaction with the smart phone 3, which is used to store calibration curves of different substances and analyze and organize the data uploaded by the smart phone. Or, as an optional alternative, the database storing the calibration curve and the software module for analyzing and sorting the data can also be directly built into the local storage device of the mobile phone.

Based on the above-mentioned detection system, the dynamic light regulation biological colorimetric detection method used in combination with mobile phones and tablet computers in the present invention is to use the smart tablet computer 2 as the bottom light source of the microtiter plate in a completely dark environment, and use the smart phone 7 as the control terminal device Realize control of light source, acquisition of color contrast images, data processing analysis and result display;

Described detection method specifically comprises the following steps:

(1) The interaction between the smart phone 7 and the smart tablet computer 2 is realized through wireless communication, and the smart phone 7 sends a light control command to the smart tablet computer 2 according to the type selection of the microtiter plate 3 and the type of the substance to be detected to control the latter to emit light range, color and brightness to form a light source dot matrix corresponding to each hole on the microtiter plate 3;

The microtiter plate 3 includes several holes arranged in an array, each hole has a transparent bottom plate and a black upper structure, and the connection between each hole is also black; the light source lattice formed by the smart tablet computer 2 and the microtiter plate 3 The transparent bottom plates of the holes correspond one by one. The type selection of the microtiter plate 3 is manually input through the operation interface of the smart phone 7 , or is automatically identified and input after taking a picture of the smart phone 7 ;

(2) After the smart tablet computer 2 completes the designated light source display, it sends a ready message to the smart phone 7, and the smart phone 7 automatically takes pictures of the microtiter plate 3 to obtain a colorimetric image;

(3) Deriving the RGB and HSL channels of each hole from the acquired colorimetric image, using the adaptive threshold algorithm to convert the colorimetric image into a binary image; then determine the positions of all circles in the image through Hough circle transformation, and obtain The center information of each hole; calculate the average pixel intensity of the 11×11 center pixel array of each hole, compare the calculation result with the standard marker curve of the detected object in the database, obtain the detection result of each hole and display it on the smartphone 7 display screen;

(4) The smart phone 7 automatically uploads the detection results to the cloud server, and the cloud server performs data sorting and analysis according to preset rules. The database storing the calibration curve can be set on the cloud server, and can also be built in the local memory of the smart phone 7 . For example, the database and the software modules for analyzing and sorting the data are built into the local storage device of the mobile phone, and this step is directly completed independently by the smart phone 7 .

The present invention will be described below through more detailed specific implementation content:

Instrument construction:

Step 1: Cabinet Design

Existing commercially available parts and components can be used for the box body and internal components of the detection system, and there is no special requirement in the present invention. It is also possible to use SOLIDWORKS 2020 soft design and 3D print by ULTRONG3D HT-500 3D printer.

Step 2: Equipment Assembly

Mount the brackets on the rails by passing the rails through the holes in the brackets and using the four bearings to secure the two rails. A smart tablet is mounted on the base of the case, and a smartphone is placed on top of the case with the rear camera located at the through hole. The assembled device is shown in Figure 1.

Step 3: Smartphone and tablet interactive program

Install applications based on the Android operating system on the smart phone and the smart tablet to realize the interaction between the two. The smart phone sends the required light source color (RGB) and brightness information (0-255) to the smart tablet according to the characteristics of the substance to be tested, and the latter analyzes the data and changes the display according to the analysis result; after the display is stable, send the ready command To the smart phone, the smart phone automatically acquires the colorimetric image under the lighting condition.

Step 4: Processing procedures for colorimetric images

In order to perform automatic image recording, data processing and storage of experimental data and establish a calibration database, a colorimetric image processing program based on the Android operating system is built into the smartphone. The application program includes two modules of detection program and database. Of course, if considering the needs of multiple smartphones being used at the same time and/or in different places, the database and the software modules for subsequent result analysis and processing can also be placed on the cloud server together, which can reduce the burden of local storage and computing, and strengthen the central computing processing ability.

After obtaining the colorimetric image, export the RGB and HSL channels from each well of the colorimetric image. The recorded colorimetric image was converted into a binary image using an adaptive threshold algorithm, and the positions of all circles in the image were determined by Hough circle transformation to obtain the center information of each hole. Calculate the average pixel intensity for the 11 x 11 center pixel array of each well. The pixel intensity of each well of different new types of substances to be detected in the microtiter plate is obtained in the above manner. By standard curve analysis of the different substances, plot the desired pixel intensity calculated from the average pixel intensity against the analyzed concentration. Acquire data for a variety of new types of substances to be detected, and build a database on this basis. The application automatically selects the best calibration curve for each substance based on the database to construct the standard marker curve. To measure existing markers in the database, the current marker library is selected directly from the user interface for actual detection. The application automatically determines the optimum conditions for the measurement procedure and the results obtained for each well are displayed directly on the application's user interface.

Biological colorimetric detection system structure and APP application interface as an example

In order to reduce the mutual interference of light sources between different wells, a microtiter plate with black well walls was used, as shown in Figure 2a. The plate display interface consists of circles with the same number of wells as the microtiter plate, and the remaining positions are black. The position and distance of each circular light-emitting area completely correspond to the microtiter plate.

The APP main interface of the smart phone is shown in Figure 3: the APP main interface is used to select the working mode of the detection system. If there is already a database for the substance, select the corresponding database to start the test. If there is no database to be detected in the database, you need to click the Add button, and use the standard concentration solution to obtain the calibration curve, and finally add the standard curve of the substance to the database.

The smartphone APP biological colorimetric image acquisition interface is shown in Figure 4: the acquired image is displayed in real time.

The display interface of the smartphone data calibration curve is shown in Figure 5: After acquiring the images of all conditions, use the adaptive threshold algorithm to convert the recorded colorimetric image into a binary image, and determine the positions of all circles in the image through Hough circle transformation , to get the center information of each hole. Calculate the average pixel intensity for the 11 x 11 center pixel array of each well. Calculate the average value of wells under the same concentration and draw calibration curves under different conditions. The program will select the best calibration curve in different concentration ranges and display it on the calibration curve interface.

As shown in Figure 6, the concentration display interface of the smart phone APP displays the final measurement result of each well.

The performance of the instrument was characterized using a standard concentration gradient of tartrazine solution. As shown in Figure 7, there are different colorimetric imaging effects under different light intensities. As the intensity of the blue light source increases, the detection sensitivity to high-concentration tartrazine solutions increases visible to the naked eye.

Subsequently, the colorimetric images of the tartrazine solution under different light intensities of the blue light source were quantitatively analyzed. It can be seen from Figure 8a that under the same light source and the same channel, as the light intensity increases, the calibration curve shifts to the right. This means higher discrimination for low concentrations at low light intensities, and higher sensitivity for high concentrations at high light intensities. The biological colorimetric detection system can detect different color channels red, green and blue (RGB) and chroma saturation brightness (HSL) under the same light source, and automatically select the best color channel. Figure 8b shows the response of the green channel to the tartrazine solution under different blue light intensities. It can be seen that the blue channel has high sensitivity to low-concentration tartrazine solution, while the green channel has high sensitivity to high-concentration tartrazine solution, thus greatly expanding the detection range of the system. Figure 8c-d shows the detection ranges of the blue channel and the green channel under different light intensities. Under the same color channel, as the light intensity increases, the maximum detection concentration also increases. For different color channels, the detection range can play a complementary role.

The invention compares the detection performance and detection efficiency of the system with the traditional plate reader. Fig. 9a is the calibration curve of the biological colorimetric detection system and the traditional plate reader for the tartrazine solution of the same concentration gradient under the blue light source. It can be clearly found that the detection system of the present invention has a wider detection range than the traditional plate reader, and the detection range is quantified in Figure 9b, which further verifies the conclusion.

At the same time, the detection time of the detection system and the traditional plate reader was calibrated. A complete inspection includes data recording and analysis. After the colorimetric image is acquired in the present invention, the program will automatically process the colorimetric image and automatically draw a calibration curve, while the data acquired by the plate reader needs to be manually counted and draw a calibration curve. As shown in Figure 10, the biocolorimetric detection system takes less than 2 seconds for one experiment, while the plate reader takes more than 200 seconds for one experiment.

The above is under the same light source, by changing the light source intensity and analyzing different color channels, the detection range of the system can be greatly expanded. At the same time, the present invention can also flexibly switch the light source according to the needs of detecting substances. Figure 11 shows the detection performance of amaranth solution under green light source and red light source. Directly observe Figures 11a and 11c, it can be found that green light source has better detection performance for amaranth solution, while red light source has better detection performance for high-concentration The first come red solution has a high sensitivity. Then, the detection performance under different light sources is also quantitatively analyzed in Figures 11b and 11d. The results showed that the detection range under the green light source was 0.001mg/mL–1.25mg/mL, and the detection range under the red light source was 0.156mg/mL–10mg/mL.

At the same time, the traditional plate reader was used to calibrate the above-mentioned amaranth solution, and its comparison with the calibration curve of the detection system is shown in Figure 12. Compared with CPR, the present invention has wider detection range (1.2×10 ^-3 -10mg/mL), higher sensitivity (9.4(mg/mL)-1, lower detection limit (detection range: 3× 10 ^-4 )–5 mg/mL; sensitivity: 1.8/(mg/mL) ^-1 ; detection limit: 1.04×10 ^-3 mg/mL). Thus, by using these typical pigment assays, it was demonstrated that the present invention can be effectively used in various colorimetric applications.

Detection example 1:

Phenol red is a typical acid-base indicator, liver function test reagent, and chromatographic reagent. In the present invention, the phenol red solution with a mass fraction ranging from 5.22×10 ^-4 to 0.625 mg/mL was used to verify the performance of the detection system. After drawing calibration curves for each type of light source, phenol red produced a good concentration-dependent response under blue and green light irradiation (shown in Figures 13a and 13b). The blue channel calibration curves and their phenol red detection ranges obtained under different blue light source light intensities are shown in Figures 13c and 13d. As the intensity of light exposure increased, the calibration curve and detection range shifted to higher concentrations. Similar results were also obtained for green light sources (Fig. 13e and 13f). The best calibration curve and detection range of phenol red obtained under blue light irradiation are shown in Fig. 13g and 13h. The results showed that the effective detection ranges of blue light and green light were 5.2×10 ^-4 -0.352 and 2.6×10 ^-2 -0.625 mg/mL, respectively, covering all concentrations in the determination of phenol red. Therefore, the present invention can be used in biomedical detection based on phenol red.

Detection example 2:

CCK-8 assay is a common cell viability bioassay widely used in anticancer drug screening and cell proliferation studies as well as cytotoxicity and drug efficacy assays. In this work, various cell samples were treated with CCK-8 reagent and then detected by the system of the present invention. The obtained results show that CCK-8 is sensitive to green light, which is consistent with the absorption peak (450nm) obtained by the plate reader. Figure 14a shows a typical image of a CCK-8 cell viability bioassay recorded under a green light source according to the present invention. As the number of cells increased, the color of the CCK-8 solution in the wells changed from green to yellow. Furthermore, the CCK-8 calibration curve shifted towards higher cell numbers with increasing light intensity (Fig. 14b). At the same time, the present invention comprehensively determined the detection range, goodness-of-fit (R ² ) value and detection sensitivity of the calibration curve (Fig. 14c-e), which shows that the system of the present invention can detect CCK-8 intelligently and dynamically. Cell numbers vary from 2,500 to 50,000. Compared to conventional plate readers, the present invention is highly automated and flexible; therefore, it can be used for cell counting and related biological assays.

Detection example 3:

Bicinchoninic acid (BCA) assay is a commonly used method for protein quantification, as protein concentration is an important biomarker for many diseases. The present invention uses the BCA kit to evaluate the detection performance of STPCPR protein. To this end, BSA solutions of different concentrations (0.05-0.75 mg/mL) were added to the BCA kit, and the resulting colorimetric images were recorded under green light illumination by the detection system (Fig. 15a). It was found that the BSA-BCA solution absorbs green light, and the higher the concentration of BCA, the lighter the color. Furthermore, the green channel shows the best detection performance among RGB and HSL channels. Figures 15b and 15c show the BSA calibration curves obtained for the green channel and their detection ranges at various green light intensities. They showed that at 2% green light intensity, the present invention exhibited good detection performance (detection range: 0.05–0.75 mg/mL, goodness-of-fit R2: ^0.9945 , sensitivity: 182(mg/mL) ^-1 ). Also, it might detect higher BSA concentrations at greater light intensities. The obtained BCA assay results show that the present invention can be used as a reliable protein detection platform.

Claims (9)

1. A dynamic light control biological colorimetric detection method used in conjunction with a mobile phone and a tablet computer, characterized in that the smart tablet computer is used as the bottom light source of the microtiter plate in a completely dark environment, and the smart phone is used as a control terminal device to realize regulation Light source, acquisition of color contrast images, data processing analysis and result display; Described detection method specifically comprises the following steps: (1) The interaction between the smart phone and the smart tablet is realized through wireless communication, and the smart phone sends light control instructions to the smart tablet according to the type of microtiter plate and the type of the substance to be detected to control the range, color and color of the latter’s light. Brightness, forming a light source dot matrix corresponding to each well on the microtiter plate; (2) After the smart tablet computer completes the display of the specified light source, it sends a ready message to the smart phone, and the smart phone automatically takes pictures of the microtiter plate to obtain a colorimetric image; (3) Deriving the RGB and HSL channels of each hole from the acquired colorimetric image, using the adaptive threshold algorithm to convert the colorimetric image into a binary image; then determine the positions of all circles in the image through Hough circle transformation, and obtain The center information of each hole; calculate the average pixel intensity of the 11×11 center pixel array of each hole, compare the calculation result with the standard marker curve of the detected object in the database, obtain the detection result of each hole and display it on the smartphone display. 2. The method according to claim 1, further comprising step (4): the smart phone automatically uploads the detection results to the cloud server, and the cloud server performs data sorting and analysis according to preset rules. 3. method according to claim 1, it is characterized in that, the selection of described microtiter plate is manually input by the operation interface of smart mobile phone, or input after automatic recognition after taking pictures by smart mobile phone; Described detected substance kind It is manually input through the operation interface of the smartphone. 4. The method according to claim 1, wherein the microtiter plate comprises several holes arranged in an array, each hole has a transparent bottom plate and a black upper structure, and the connecting portion between each hole is also It is black; the light source dot matrix formed by the smart tablet corresponds to the transparent bottom plate of each well on the microtiter plate. 5. The method according to claim 1, wherein the database is set on a cloud server, or built into a local memory of the smart phone. 6. A dynamic light control biological colorimetric detection system used in combination with a mobile phone and a tablet computer, characterized in that it includes a fully enclosed and light-tight box structure; The tablet computer, the microtiter plate is suspended above the smart tablet computer by the bracket and keeps an appropriate distance; there is a through hole on the top of the box, and the smart phone is placed on the top of the box with the display screen upward, and can use the through hole and the rear The camera shoots the microtiter plate; The smart phone has a built-in image detection, processing and analysis module, which is used for acquiring colorimetric images, data processing and analysis and displaying results according to the method described in claim 1. 7. The system according to claim 6, characterized in that, the system also includes a cloud server capable of data interaction with smart phones, for storing calibration curves of different substances and/or analyzing data uploaded by smart phones and finishing. 8. The system according to claim 6, wherein the support is a drawable guide rail support, including four bearings arranged on the box body, and every two bearings form a group and are respectively connected by guide rails, The two guide rails are arranged in parallel; the bracket rests on the guide rails, and the microtitration plate is embedded in the mounting position of the bracket. 9. The system according to claim 6, wherein the aperture of the through hole on the top of the box is 3cm, and the distance between the rear camera of the smart phone and the microtiter plate is 30cm.

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