A cholesterol measurement system based on RGB color detection

Abstract

The present invention discloses a cholesterol measurement system based on RGB color detection, which includes a light-shielding darkroom, a microprocessor, a display component, and a detection component placed in the light-shielding darkroom. The detection component includes a support frame, on which a detection cell for placing a cuvette is provided, and a white balance light source, an RGB color sensor, and an excitation light source are further arranged thereon. The installation centerlines of the white balance light source, the RGB color sensor, and the excitation light source are on the same horizontal plane; wherein the white balance light source and the RGB color sensor are respectively arranged on both sides of the detection cell facing each other directly, and the excitation light source is arranged on the other side of the detection cell. The lights emitted by the white balance light source and the excitation light source irradiate on the cuvette, and the RGB color sensor collects the fluorescence intensity of the solution in the cuvette as an input; the microprocessor is respectively connected to the RGB color sensor and the display component. The measurement system of the present invention is simple and easy to operate, has high sensitivity and high accuracy.

Description

Technical Field

[0001] The present invention relates to a cholesterol measurement system based on RGB color detection, belonging to the technical field of biological detection. Background Art

[0002] Cholesterol is an important lipid in humans and is a major component of cell membranes, helping to maintain membrane permeability and fluidity. At normal concentrations in the human body, cholesterol plays an important role in the digestion of food. Cholesterol plays four different important roles in the human body, namely forming digestive bile acids in the intestine, producing vitamin D, forming cell membranes and some hormones. Excessive cholesterol content in human serum will form plaques in blood vessel channels, thus hindering blood circulation and causing cardiovascular diseases; while too low cholesterol content will also induce diseases such as cancer, Alzheimer's disease and cerebral hemorrhage. Therefore, cholesterol has become an important biomarker for many diseases and is also one of the most common detection items in clinical practice.

[0003] There are more than 200 methods for measuring total cholesterol, which can be divided into four categories: chemical reagent colorimetry, enzyme analysis, fluorescence method, and high performance liquid chromatography. In recent years, the fluorescence method has played an increasingly important role in the field of bioanalysis research. At present, there are relevant reports on the application of gold nanoparticles modified with β-cyclodextrin (Beta cyclodextrin, β-CD) in the fluorescence method for detecting cholesterol content in human serum.

[0004] The previous research results of the inventor team of this application (publication number: CN113801650A) show that the thiol-β-cyclodextrin-gold nanocluster solution itself has fluorescence properties (emitting green fluorescence) under ultraviolet light. After the thiol-β-cyclodextrin-gold nanocluster contacts with cholesterol, the fluorescence intensity of the thiol-β-cyclodextrin-gold nanocluster can be further enhanced, which can be used for the detection of cholesterol content in human serum, and has strong anti-interference ability and high sensitivity. When the cholesterol concentration is 10.0~100.0 μmol·L -1 , the cholesterol concentration has a linear relationship with the fluorescence enhancement degree of the thiol-β-cyclodextrin-gold nanocluster. Based on the linear relationship between the cholesterol concentration and the fluorescence enhancement degree of the thiol-β-cyclodextrin-gold nanocluster, the inventor team of this application further studied that the fluorescence color of the thiol-β-cyclodextrin-gold nanocluster solution added with cholesterol is linearly related to the cholesterol concentration, and thus established the cholesterol detection system based on RGB color detection of this application. Summary of the Invention

[0005] The technical problem to be solved by the present invention is to provide a simple and easy-to-operate cholesterol measurement system based on RGB color detection with high sensitivity and high accuracy.

[0006] To solve the above technical problems, the present invention adopts the following technical solutions:

[0007] A cholesterol measurement system based on RGB color detection, comprising a light-shielding darkroom, a microprocessor, a display component, and a detection component disposed in the light-shielding darkroom; wherein:

[0008] The light-shielding darkroom is composed of a receiving cavity and a cover plate;

[0009] The detection component includes a support frame, on which a detection cell for placing a colorimetric cuvette is provided, and a white balance light source, an RGB color sensor, and an excitation light source are further provided on the support frame. The installation center lines of the white balance light source, the RGB color sensor, and the excitation light source are on the same horizontal plane; wherein the white balance light source and the RGB color sensor are respectively disposed oppositely and directly on both sides of the detection cell, the excitation light source is disposed on the other side of the detection cell, and the light emitted by the white balance light source and the excitation light source irradiates on the colorimetric cuvette, and the RGB color sensor collects the fluorescence intensity of the solution in the colorimetric cuvette as an input;

[0010] The RGB color sensor is connected to the microprocessor, and the microprocessor is connected to the display component.

[0011] Further, the cholesterol measurement system based on RGB color detection of the present invention further includes a power supply module for supplying power to components such as the RGB color sensor and the microprocessor, which are respectively connected to the RGB color sensor and the microprocessor. Specifically, the power supply module can be a lithium battery. Still further, the cholesterol measurement system further includes a host computer for processing and storing various data (including the cholesterol concentration-hue value standard curve, the hue values of various solutions, etc.), and the host computer is respectively connected to the microprocessor and the display component.

[0012] In the technical solution of the present invention, the white balance light source is preferably a white LED light source; the excitation light source is preferably an ultraviolet lamp, and further preferably an ultraviolet lamp with a wavelength of 365 nm or 370 nm.

[0013] In the technical solution of the present invention, the RGB color sensor is an RGB color sensor that satisfies the input of fluorescence intensity and outputs R frequency, G frequency, and B frequency, and the preferred model is the TCS230 RGB color sensor.

[0014] In the technical solution of the present invention, the display component is a display screen, specifically an LCD touch display screen.

[0015] The present invention also provides a method for measuring cholesterol concentration using the above system, comprising the following steps:

[0016] 1) Place the mercapto-β-cyclodextrin-gold nanocluster solution (also simply referred to as CD-AuNCs solution in this application) in a cuvette, add a cholesterol standard solution with a certain concentration. After the reaction is completed, place the cuvette in the detection pool on the detection component in a light-shielded darkroom, and cover the cover plate;

[0017] 2) Turn on the white balance light source, adjust the white balance of the RGB color sensor and then turn off the white balance light source. Then turn on the excitation light source. Under the irradiation of the excitation light source, the RGB color sensor collects the fluorescence intensity of the solution in the cuvette to obtain the fluorescence color of the solution in the cuvette;

[0018] 3) The fluorescence color of the solution in the obtained cuvette is processed by the microprocessor, converted into the HSV color space, and the hue value is recorded to obtain the hue value corresponding to the current cholesterol standard solution concentration;

[0019] 4) According to the order of steps 1) to 3), detect multiple cholesterol standard solutions with different concentrations, and obtain multiple hue values corresponding to the cholesterol standard solution concentrations;

[0020] 5) Use the cholesterol concentration in the cholesterol standard solution and the corresponding hue value to make a relationship curve to obtain a cholesterol concentration-hue value standard curve. Input this standard curve into the microprocessor and display it through the display component;

[0021] 6) Repeat steps 1) to 3), replace the cholesterol standard solution with the serum to be tested, and obtain the hue value corresponding to the cholesterol concentration in the serum to be tested;

[0022] 7) According to the cholesterol concentration-hue value standard curve, obtain the cholesterol concentration corresponding to the hue value described in step 6) and display it on the display component. This cholesterol concentration is the cholesterol concentration in the serum to be tested.

[0023] The CD-AuNCs solution involved in the present invention is preferably prepared by the method described in the patent application with the publication number CN113801650A.

[0024] In step 1) of the above method, in the detection system, the concentration of mercapto-β-cyclodextrin-gold nanocluster in the system is preferably 1.9×10 -5 ~5.8×10 -5 mol·L -1 (calculated by Au). If the concentration is too high, it can be diluted with PBS buffer solution with pH = 6.4 as the solvent or dispersant. In this step, the reaction time of CD-AuNCs and cholesterol is usually greater than or equal to 8 min, preferably 8 min.

[0025] Compared with the prior art, the measurement system of the present invention uses a TCS230RGB color sensor to detect the fluorescence color of the solution after the reaction of CD-AuNCs and cholesterol, obtains the values of the three primary colors of RGB, and then converts them into the HSV color space through the processing of a microprocessor. The applicant has found that the hue value in the HSV color space is negatively correlated with the cholesterol concentration corresponding to this hue value and has a good linear relationship. Therefore, a cholesterol concentration-hue value standard curve can be constructed. Then, by obtaining the hue value of the solution after the reaction of the serum to be measured and the CD-AuNCs solution and combining it with the above standard curve, the cholesterol concentration in the serum to be measured can be obtained. The measurement system of the present invention is simple to operate, highly sensitive, and highly accurate. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 It is an assembly structure diagram of an embodiment of the cholesterol measurement system based on RGB color detection of the present invention.

[0027] Figure 2 is Figure 1 A schematic structural diagram of the detection component in the shown embodiment.

[0028] Figure 3 It is a cholesterol concentration-hue value standard curve fitted by using the system and method of the present invention.

[0029] The reference numerals in the figure are:

[0030] 1 Cover plate, 2 Support frame, 3 White balance light source, 4 Excitation light source, 5 Accommodation cavity, 6 Base, 7 Concave cavity, 8 Microprocessor, 9 Display component, 10 RGB color sensor, 11 Detection cell, 12 Colorimetric cuvette. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0031] In order to better explain the technical solution of the present invention, the present invention will be further described in detail below with reference to the accompanying drawings, but the embodiments of the present invention are not limited thereto.

[0032] A cholesterol measurement system based on RGB color detection according to the present invention includes a light-shielding darkroom, a microprocessor 8, a display component 9, and a detection component placed in the light-shielding darkroom. The detection process is carried out in the light-shielding darkroom. The data obtained from the detection is sent to the microprocessor 8 for processing, and the results are displayed on the display component 9. Specifically, the fluorescence color of the solution in the cuvette 12 is obtained through detection. The microprocessor 8 is used to implement the processing and recording of various data. For example, the fluorescence color transmitted by the RGB color sensor 10 is converted into the HSV color space, and the hue value therein is recorded; the cholesterol concentration-hue value standard curve is fitted and recorded. The display component 9 is used to display various data, including the cholesterol concentration-hue value standard curve, the hue value, the cholesterol concentration value in the serum to be measured, etc. The light-shielding darkroom, the microprocessor 8, and the display component 9 can be simultaneously arranged on the same base 6, as Figure 1 shown; or the light-shielding darkroom can be separately arranged from the microprocessor 8 and the display component 9, that is, the light-shielding darkroom is an independent component, and the microprocessor 8 and the display component 9 can be arranged together or separately.

[0033] In Figure 1 the shown embodiment, the light-shielding darkroom, the microprocessor 8, and the display component 9 are simultaneously arranged on a base 6. An accommodation cavity 5 and a concave cavity 7 are formed on the base 6. A cover plate 1 matching the accommodation cavity 5 is provided on the accommodation cavity 5. The accommodation cavity 5 and the cover plate 1 form the light-shielding darkroom, and the detection component is placed in the light-shielding darkroom. The microprocessor 8 and the display component 9 are arranged on the same circuit board, and they are placed in the concave cavity 7 on the base 6. The size of the display component 9 just covers the opening of the concave cavity 7, or is slightly larger than the opening of the concave cavity 7. The output end of the RGB color sensor 10 is connected to the input end of the microprocessor 8, and the human-computer interaction data control end of the microprocessor 8 is connected to the display component 9.

[0034] The structure of the detection component is as Figure 2As shown in the figure, it includes a support frame 2. A detection cell 11 for placing a colorimetric cuvette 12 (the colorimetric cuvette 12 is used to hold a cholesterol standard solution or a test serum containing cholesterol and a CD-AuNCs solution for reaction) is provided on the support frame 2. A white balance light source 3, an RGB color sensor 10, and an excitation light source 4 are also provided on the support frame 2. The white balance light source 3 is used to adjust the white balance of the RGB color sensor 10. The excitation light source 4 is used to excite the solution in the colorimetric cuvette 12 to produce a fluorescence color. The RGB color sensor 10 is used to collect the fluorescence intensity generated by the solution in the colorimetric cuvette 12 under the irradiation of the excitation light source 4, so as to obtain the fluorescence color (R, G, B values) of the solution in the colorimetric cuvette 12. The white balance light source 3, the RGB color sensor 10, and the excitation light source 4 all need to face the colorimetric cuvette 12 to obtain accurate data, and the irradiation components of the white balance light source 3 and the excitation light source 4 should be consistent with the part where the RGB color sensor 10 collects information. Therefore, the installation center lines of the white balance light source 3, the RGB color sensor 10, and the excitation light source 4 should be on the same horizontal plane. The detection optical path is an orthogonal optical path, that is, the excitation light emitted by the excitation light source 4 and the emission light emitted by the solution in the colorimetric cuvette 12 are at 90°. Specifically, the white balance light source 3 and the RGB color sensor 10 can be respectively arranged on both sides of the detection cell 11 facing each other, and the excitation light source 4 is arranged on the other side of the detection cell 11. The light emitted by the white balance light source 3 and the excitation light source 4 is directly irradiated on the colorimetric cuvette 12 in the detection cell 11. The RGB color sensor 10 collects the fluorescence intensity generated by the solution in the colorimetric cuvette 12 under the excitation light source 4 as the input, and its output is the fluorescence color of the solution in the colorimetric cuvette 12 under the irradiation of the excitation light source 4. For the RGB color sensor 10, it should satisfy that the input is the fluorescence intensity and the output is the RGB color sensor 10 of the R frequency, G frequency, and B frequency. Figure 3 is Figure 1 the module block diagram of the cholesterol measurement system based on RGB color detection shown in the figure.

[0035] Furthermore, the cholesterol measurement system based on RGB color detection of the present invention further includes a power supply module for supplying power to components such as the RGB color sensor 10 and the microprocessor 8, which are respectively connected to the RGB color sensor 10 and the microprocessor 8. Specifically, the power supply module can be a lithium battery or other DC power supplies that meet the requirements.

[0036] Even further, the cholesterol measurement system based on RGB color detection of the present invention may further include a host computer for realizing human-computer interaction and displaying various data (including the cholesterol concentration-hue value standard curve, the hue values of various solutions, etc.). The host computer is respectively connected to the microprocessor 8 and the display component 9.

[0037] In the technical solution of the present invention, the microprocessor 8 is also called a microcontroller unit (MCU) or a single-chip microcomputer, and its selection is an existing conventional selection that can implement data processing. The display component 9 is a display screen, specifically an LCD touch display screen.

[0038] In a specific embodiment, the RGB color sensor 10 preferably adopts a TCS230RGB color sensor 10, and the model of the microprocessor 8 is STM32F407.

[0039] In a specific embodiment, the white balance light source 3 is a white LED light source; the excitation light source 4 is an ultraviolet lamp, and an ultraviolet lamp with a wavelength of 365 nm or 370 nm is further adopted.

[0040] The present invention also provides a method for measuring cholesterol concentration using the above system, including the following steps:

[0041] 1) Take the CD-AuNCs solution and place it in the cuvette 12, add a cholesterol standard solution with a certain concentration. After the reaction is completed, place the cuvette 12 in the detection cell 11 on the detection component in a light-shielded darkroom, and cover the cover plate 1;

[0042] 2) Turn on the white balance light source 3, adjust the white balance of the RGB color sensor 10 and then turn off the white balance light source 3. Then turn on the excitation light source 4. Under the irradiation of the excitation light source 4, the RGB color sensor 10 collects the fluorescence intensity of the solution in the cuvette 12 to obtain the fluorescence color of the solution in the cuvette 12;

[0043] 3) The fluorescence color of the solution in the obtained cuvette 12 is processed by the microprocessor 8, converted into the HSV color space, and the hue value is recorded to obtain the hue value corresponding to the current cholesterol standard solution concentration;

[0044] 4) According to the order of steps 1) to 3), detect multiple cholesterol standard solutions with different concentrations to obtain multiple hue values corresponding to the cholesterol standard solution concentrations;

[0045] 5) Use the cholesterol concentration in the cholesterol standard solution and the corresponding hue value to make a relationship curve to obtain a cholesterol concentration - hue value standard curve. Input this standard curve into the microprocessor 8 and display it through the display component 9;

[0046] 6) Repeat steps 1) to 3), replace the cholesterol standard solution with the serum to be tested, and obtain the hue value corresponding to the cholesterol concentration in the serum to be tested;

[0047] 7) According to the cholesterol concentration - hue value standard curve, obtain the cholesterol concentration corresponding to the hue value described in step 6) and display it on the display component 9. This cholesterol concentration is the cholesterol concentration in the serum to be tested.

[0048] In the present invention, the CD-AuNCs solution involved is preferably prepared according to the method described in the patent application with the publication number CN113801650A. Specifically as follows:

[0049] Preparation of CD-AuNCs solution: Synthesized by the reduction method. The specific synthesis steps are as follows: All glassware is soaked and washed clean with freshly prepared aqua regia. Adjust the water bath temperature to 90 °C. Take 5.0 mL of HAuCl4·3H2O aqueous solution (1.0 mM, 90 °C) and stir vigorously for 10 min, then add 2.0 mL of mono(6-mercapto-6-deoxy)beta-cyclodextrin solution (20.0 mM, 90 °C). After stirring for 5 min, add 1.5 ml of sodium hydroxide aqueous solution (1.0 mol·L -1 ), then seal the bottle mouth with an aluminum film and react for 4 h. The solution is a yellow clear liquid, and a strong green fluorescence can be observed under ultraviolet light (365 nm), indicating that a green light CD-AuNCs solution is obtained. The concentration of CD-AuNCs in the solution is 5.8×10 -4 mol·L -1 .

[0050] Purification of CD-AuNCs: Put the CD-AuNCs solution into a dialysis bag with a cut-off molecular weight of 35 KDa and dialyze in distilled water for 2 days, changing the distilled water every 4 hours. The dialyzed solution is freeze-dried and stored at 4 °C.

[0051] In the method of the present invention, for the CD-AuNCs solution placed in the cuvette 12, the concentration of CD-AuNCs is preferably 1.9×10 -5 ~5.8×10 -5 mol·L -1 (calculated as Au). If the concentration is too high, it can be diluted with PBS buffer solution with pH = 6.4 as the solvent or dispersant.

[0052] In the method of the present invention, after 8 min of reaction between the cholesterol standard solution or the serum to be tested containing cholesterol and CD-AuNCs, the fluorescence intensity remains basically unchanged. Therefore, the reaction time usually needs to be greater than or equal to 8 min, preferably 8 min.

[0053] In the method of the present invention, the number of portions of the cholesterol standard solution can be determined as needed. In order to establish a quantitative relationship between the enhancement of CD-AuNCs and the cholesterol concentration, 7 cholesterol solutions with different concentrations (0, 20, 40, 60, 90, 120, 150 μmol·L -1 ) are respectively prepared within the concentration range of 10.0~150.0 μmol·L -1)。Take 50.0 μL of cholesterol at the above concentration and add it dropwise to 1.0 mL of the CD-AuNCs solution prepared by the aforementioned method. The reaction temperature is 25 °C and the reaction time is 8 min. Then, measure its fluorescence emission spectrum and fluorescence intensity (fluorescence color), and convert it into the HSV color space. The fluorescence color and the parameter values of the HSV color space are shown in Table 1 below.

[0054] Table 1:

[0055]

[0056] The experimental results show that:

[0057] (1) The addition of cholesterol can enhance the fluorescence intensity of the solution, and as the cholesterol concentration increases, the fluorescence intensity of the solution increases.

[0058] (2) The addition of cholesterol can reduce the hue value (H) of the HSV color space of the solution, and as the cholesterol concentration increases, the hue value of the solution decreases.

[0059] Using the cholesterol concentration values in the cholesterol standard solution in Table 1 and the corresponding hue values H of the HSV color space to make a correlation curve, the correlation coefficient between the two reaches 0.97, as specifically shown in Figure 3 shown. It can be seen that the system described in the present invention has good accuracy and stability, the detection results can be displayed in real time, and rapid quantitative detection of cholesterol can be achieved.

Claims

1. A cholesterol measurement system based on RGB color detection, comprising a light-shielding darkroom, a microprocessor (8), a display component (9), and a detection component placed in the light-shielding darkroom. It is characterized in that, The light-shielding darkroom is composed of a receiving cavity (5) and a cover plate (1); The detection component includes a support frame (2). A detection pool (11) for placing a cuvette (12) is provided on the support frame (2). The cuvette (12) is filled with a mercapto-β-cyclodextrin-gold nanocluster solution. A white balance light source (3), an RGB color sensor (10), and an excitation light source (4) are also provided on the support frame (2). The installation centerlines of the white balance light source (3), the RGB color sensor (10), and the excitation light source (4) are on the same horizontal plane. Among them, the white balance light source (3) and the RGB color sensor (10) are respectively arranged oppositely and directly on both sides of the detection pool (11), and the excitation light source (4) is arranged on the other side of the detection pool (11). The light emitted by the white balance light source (3) and the excitation light source (4) irradiates on the cuvette (12), and the RGB color sensor (10) collects the fluorescence intensity of the solution in the cuvette (12) as input. Among them, the white balance light source (3) is a white LED light source, and the excitation light source (4) is an ultraviolet lamp with a wavelength of 365 nm or 370 nm; The RGB color sensor (10) is connected to the microprocessor (8), and the microprocessor (8) is connected to the display component (9); the fluorescence color of the solution in the cuvette (12) is processed by the microprocessor (8), converted into the HSV color space, and the hue value therein is recorded to obtain the hue value corresponding to the cholesterol solution concentration.

2. The cholesterol measurement system based on RGB color detection according to claim 1, characterized in that, This cholesterol measurement system further includes a power supply module, and the power supply module is respectively connected to the RGB color sensor (10) and the microprocessor (8).

3. The cholesterol measurement system based on RGB color detection according to claim 1, characterized in that, This cholesterol measurement system further includes a host computer, and the host computer is respectively connected to the microprocessor (8) and the display component (9).

4. The cholesterol measurement system based on RGB color detection according to any one of claims 1 to 3, characterized in that, The model of the RGB color sensor (10) is TCS230.

5. The cholesterol measurement system based on RGB color detection according to any one of claims 1 to 3, characterized in that, The display component (9) is a display screen.

6. A method for measuring cholesterol concentration using the cholesterol measurement system based on RGB color detection according to claim 1, comprising the following steps: 1) Take the mercapto-β-cyclodextrin-gold nanocluster solution and place it in the cuvette (12), add a cholesterol standard solution with a certain concentration. After the reaction is completed, place the cuvette (12) in the detection pool (11) on the detection component in the light-shielding darkroom, and cover the cover plate (1); 2) Turn on the white balance light source (3), adjust the white balance of the RGB color sensor (10), then turn off the white balance light source (3), and then turn on the excitation light source (4). Under the irradiation of the excitation light source (4), the RGB color sensor (10) collects the fluorescence intensity of the solution in the cuvette (12) to obtain the fluorescence color of the solution in the cuvette (12); 3) The obtained fluorescence color of the solution in the cuvette (12) is processed by the microprocessor (8), converted into the HSV color space, and the hue value therein is recorded to obtain the hue value corresponding to the current cholesterol standard solution concentration; 4) Detect cholesterol standard solutions with different concentrations in the order of steps 1) to 3) to obtain hue values corresponding to the concentrations of the cholesterol standard solutions; 5) Use the cholesterol concentration in the cholesterol standard solution and the corresponding hue value to make a relationship curve to obtain a cholesterol concentration - hue value standard curve. Input this standard curve into the microprocessor (8) and display it through the display component (9); 6) Repeat steps 1) to 3), replacing the cholesterol standard solution with the serum to be tested, to obtain a hue value corresponding to the cholesterol concentration in the serum to be tested; 7) According to the cholesterol concentration - hue value standard curve, obtain the cholesterol concentration corresponding to the hue value described in step 6) and display it on the display component (9). This cholesterol concentration is the cholesterol concentration in the serum to be tested.

7. The method according to claim 6, characterized in that, In step 1), the reaction time is greater than or equal to 8 min.

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