Fluorescent quantification apparatus

Abstract

The utility model relates to fluorescent detection technical field especially relates to fluorescent quantitative analysis equipment, including base, still including light -proof door and object table, the upper end fixedly connected with instrument light -proof cover and handheld type charging ultraviolet lamp of base, the upper side of instrument light -proof cover front end is equipped with image acquisition chamber, the front end of image acquisition chamber is provided with light -proof door, the upper end of image acquisition chamber inner wall is slidably connected with object table, and the upper end of object table is provided with positioning rivet, and the front end fixedly connected with light -proof plate of object table, the inside of image acquisition chamber is provided with camera body, the utility model discloses through the use of light -proof door and object table, will positioning rivet install on object table, and the flexibility of replacement reduces the equipment cost, improves the versatility and practicality of equipment, places the uric acid color paper core piece on positioning rivet, and drops the fluorescence sample, and the object table is slid into image acquisition chamber through light -proof door, and the light -proof environment of sealed auxiliary formation of light -proof plate avoids the interference of outside light source to the image.

Description

Technical Field

[0001] This utility model relates to the field of fluorescence detection technology, and in particular to fluorescence quantitative analysis equipment. Background Technology

[0002] Fluorescence quantitative analysis equipment is a type of precision instrument used to detect and quantify fluorescence signals in samples. It achieves quantitative detection of fluorescent substances through ultraviolet excitation and image analysis. Microfluidic paper chip technology has shown broad application prospects in fields such as biological detection and medical diagnosis due to its advantages such as simple operation, low cost and automation.

[0003] Existing quantitative fluorescence analysis equipment has a relatively simple structure and cannot be adapted to paper chip carriers of different shapes. This results in insufficient sample compatibility, increases equipment costs, and the equipment has poor light shielding effect, making it susceptible to ambient light and interfering with the accuracy of image acquisition, thus affecting the precision and efficiency of quantitative fluorescence analysis.

[0004] Therefore, to address the issues of insufficient sample compatibility and poor light-shielding effect of existing fluorescence quantitative analysis equipment, which affect the accuracy of fluorescence quantitative analysis, a fluorescence quantitative analysis device can be designed. Utility Model Content

[0005] To overcome the problems of insufficient sample compatibility and poor light-shielding effect of existing fluorescence quantitative analysis equipment, which affect the accuracy of fluorescence quantitative analysis.

[0006] The technical solution of this utility model is as follows: a fluorescence quantitative analysis device, including a base; it also includes a light-shielding door and a stage. An instrument light-shielding cover and a handheld rechargeable ultraviolet lamp are fixedly connected to the upper end of the base. An image acquisition cavity is opened on the upper side of the front end of the instrument light-shielding cover. A light-shielding door is set at the front end of the image acquisition cavity. A stage is slidably connected to the upper end of the inner wall of the image acquisition cavity. A positioning rivet is set at the upper end of the stage. A light shield is fixedly connected to the front end of the stage. A camera body is set inside the image acquisition cavity.

[0007] Preferably, the camera body focal length is manually adjusted to clearly acquire the fluorescence image on the paper chip. The handheld rechargeable ultraviolet lamp is turned on, the light-shielding door is installed, the matching software is opened, the standard curve parameters, reaction time, and automatic recognition template are set, the positioning rivet is installed on the stage, the uric acid colorimetric paper chip is placed on the positioning rivet, and the fluorescent sample is added. The start detection button on the experiment page is clicked, and the stage is slid into the image acquisition cavity through the light-shielding door. The camera body automatically acquires the fluorescence image after the fluorescent substance on the chip is excited and transmits the data. The final color value and concentration are recorded in the software's historical data module.

[0008] As a preferred option, the instrument's light shield, light shield door, stage, positioning rivets, and light shield are all made of black resin material designed using 3D modeling.

[0009] Preferably, the instrument's light shield has a UV lamp inlet at the rear end, which is compatible with the front end of the handheld rechargeable UV lamp. The right end of the handheld rechargeable UV lamp is fixedly connected to a cable interface.

[0010] Preferably, the surface of the light-proof door is provided with mounting holes and an observation window, and the rear end of the light-proof door is slidably connected to a light-blocking window, which is compatible with the observation window.

[0011] Preferably, a limiting seat is fixedly connected to the upper end of the inner wall of the image acquisition cavity, and the two ends of the stage are slidably connected along the limiting seat and the mounting hole, with the light shield being larger than the mounting hole.

[0012] Preferably, the camera body is a 4K macro camera, with a lens and a fill light fixedly connected to the lower end of the camera body, and the lens and the positioning rivet are located on the same axis.

[0013] Preferably, a nut is threaded onto the right end of the camera body, and the other end of the nut extends to the outside of the instrument's light shield and is threaded onto a nut.

[0014] The beneficial effects of this utility model are:

[0015] This fluorescence quantitative analysis device, through the use of a light-shielding door and a stage, allows manual adjustment of the camera's focal length to clearly acquire fluorescence images on the paper chip, improving detection accuracy. A handheld rechargeable UV lamp provides a stable UV light source. The light-shielding door is installed, the accompanying software is opened, and the standard curve parameters, reaction time, and automatic recognition template are set. Positioning rivets are selected based on the shape of the uric acid chromogenic paper chip and installed on the stage. This flexible installation reduces equipment costs and improves the device's versatility and practicality. The uric acid chromogenic paper chip is placed on the positioning rivet, and a fluorescent sample is added. Clicking the "Start Detection" button on the experiment page allows the stage to slide into the image acquisition chamber through the light-shielding door. A light-shielding plate helps create a sealed, light-shielding environment, preventing interference from external light sources. The camera automatically acquires the fluorescence image after the fluorescent substance on the chip is excited and transmits the data. The final colorimetric value and concentration are recorded in the software's historical data module. Attached Figure Description

[0016] Figure 1 The diagram shown is a schematic representation of the overall three-dimensional structure of this utility model. Figure 1 ;

[0017] Figure 2 The diagram shown is a schematic representation of the overall three-dimensional structure of this utility model. Figure 2 ;

[0018] Figure 3 The diagram shown is a three-dimensional cross-sectional view of the present invention.

[0019] Figure 4 The diagram shown is a schematic representation of the platform structure of this utility model.

[0020] Figure 5 The diagram shown is a schematic representation of the image acquisition cavity structure of this utility model.

[0021] Explanation of reference numerals in the attached diagram: 1. Base; 2. Instrument light shield; 3. Handheld rechargeable UV lamp; 4. Image acquisition cavity; 5. Light shield door; 6. Stage; 7. Positioning rivet; 8. Light shield plate; 9. Camera body; 10. UV lamp socket; 11. Cable interface; 12. Mounting hole; 13. Observation window; 14. Light shield window; 15. Limiting seat; 16. Lens; 17. Fill light; 18. Nut; 19. Screw cap. Detailed Implementation

[0022] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0023] To make the above-mentioned objects, features and advantages of the present invention more apparent and understandable, the present invention will be further described in detail below with reference to the accompanying drawings and specific embodiments.

[0024] In the field of modern analytical testing, with the deep integration of biotechnology, materials science, and information technology, the demand for rapid, accurate, and low-cost testing is becoming increasingly urgent. Microfluidic paper-based chips, as an emerging analytical testing method, are gaining prominence in multiple fields due to their unique advantages. Using paper as a substrate, they leverage the paper's natural porous structure and capillary action to achieve automated sample transport and reaction, eliminating the need for complex external pumping systems and significantly reducing equipment costs. Simultaneously, paper-based materials possess excellent biocompatibility, adaptable to various biological samples such as blood, saliva, and urine, facilitating early disease diagnosis, and are particularly suitable for disease screening in resource-scarce areas.

[0025] In the field of disease diagnosis, microfluidic paper chips can integrate multiple detection functions, enabling rapid detection of infectious disease pathogens, tumor markers, and more, and are expected to become a powerful tool for point-of-care testing (POCT) in primary healthcare. In environmental monitoring, they can rapidly detect heavy metal ions and organic pollutants in water, as well as harmful gases in the air, meeting the needs of emergency environmental monitoring. In food safety testing, they can quickly detect pesticide residues, veterinary drug residues, and microbial toxins in food, ensuring the safety of the food supply chain. However, microfluidic paper chip technology still faces many challenges in practical applications, especially in the supporting detection equipment. Insufficient equipment in key areas such as fluorescence excitation, image acquisition, and data processing limits the further promotion and application of this technology. Innovative equipment is urgently needed to overcome these technical bottlenecks and promote the industrialization and practical application of microfluidic paper chip technology.

[0026] Microfluidic paper chip technology has become a research hotspot in disease diagnosis, environmental monitoring, and food safety testing due to its advantages such as low cost, portability, and adaptability to biological samples. However, traditional fluorescence quantitative analysis devices have significant shortcomings: their simple structure, often designed for specific detection processes, makes it difficult to accommodate the entire process of "precise fluorescence excitation—stable image acquisition—efficient quantitative analysis" when adapted to microfluidic paper chips; insufficient light-proof environment construction allows stray light to easily interfere with fluorescence signals, leading to detection errors; poor stability of ultraviolet light sources, with fluctuations in luminescence intensity and wavelength affecting fluorescence excitation and interfering with quantitative analysis; and low software adaptability, with most devices relying on general image analysis software, making it impossible to customize data processing for the fluorescence characteristics and colorimetric experimental requirements of microfluidic paper chips.

[0027] Meanwhile, in existing technologies, similar portable multi-channel fluorescent paper-based microfluidic chips based on smartphone imaging, although attempts have been made to achieve detection by combining with portable devices, still have room for improvement in the construction of light-shielding environments, light source stability, and software quantitative accuracy.

[0028] The self-developed supporting software is deeply adapted to the hardware and has three core functions: Fluorescence-to-chromaticity value conversion: Through machine learning algorithms, a precise mapping model is established between fluorescence intensity and chromaticity values, as well as color space parameters such as RGB and HSV. For different fluorescent materials, such as FITC, Cys, different paper chip materials, cellulose filter paper, and nitrocellulose membranes, the conversion parameters are automatically calibrated to convert the fluorescence signal into a quantifiable chromaticity value, solving the error problem caused by the "one-size-fits-all" conversion of traditional software.

[0029] Concentration characterization algorithm: Based on a large amount of standard sample detection data, covering different concentration gradients and different fluorescent markers, a concentration-chromaticity value correlation model is trained. It supports linear fitting, nonlinear fitting, and various algorithms such as the Michaelis-Menten model. It can intelligently select the optimal model according to the characteristics of the detected substance, such as enzyme-catalyzed fluorescence and immunofluorescence, to achieve accurate inversion of fluorescent substance concentration. Taking the detection of glucose using a microfluidic paper chip as an example, the software can quickly calculate the glucose concentration through chromaticity values, with a detection error of <5%.

[0030] Multi-scenario Adaptability and Expansion: In addition to quantitative fluorescence analysis, the software supports the acquisition and analysis of colorimetric experimental images. For colorimetric detection methods, such as pH test paper color development and heavy metal chromogenic reagent reactions, it can identify the color intensity of RGB channels and, combined with color recognition algorithms, convert color depth into quantitative results such as concentration and pH value. For example, in the pH detection of environmental water samples, after the paper chip develops color, the software can accurately identify color changes, achieving a detection accuracy of ±0.1 for pH value.

[0031] Existing feasible technologies include image acquisition devices based on the microfluidic paper chip method. These devices are mainly used to excite fluorescent substances dropped onto a paper chip and acquire fluorescence images after excitation. When used in conjunction with supporting software, fluorescence intensity can be quantified into chromaticity values, which can then be used to characterize the concentration of fluorescent substances, thereby achieving quantitative analysis.

[0032] It is suitable for acquiring fluorescence images using the microfluidic paper chip method. It can provide a completely light-proof environment and a stable ultraviolet light source. It can then be used with self-developed software for quantitative analysis of fluorescence intensity and ultimately characterize the concentration of fluorescent substances. In addition, the device is also suitable for image acquisition in all other colorimetric experiments. With the accompanying software, it can identify color intensity and achieve accurate characterization of color intensity.

[0033] This experiment synthesized nitrogen-doped carbon quantum dots (N-CQDs) via a high-temperature hydrothermal method using citric acid as the carbon source and ethylenediamine as the nitrogen source. The amino groups (-NH2) on the surface of the prepared N-CQDs can self-assemble with gold nanoparticles (AuNPs) through N-Au coordination to form CNQDs@AuNPs. After adding aspartic acid (Asp), the CNQDs@AuNPs@Asp functionalized probes were obtained by further utilizing the -NH2 coordination of aspartic acid (Asp). The specific molecular recognition between CNQDs@AuNPs@Asp and Cys affects the fluorescence signal. Using a microfluidic paper chip as the detection platform, the results were recorded and the chromaticity values ​​were analyzed using a self-made imaging device and software. The quantitative detection of Cys was achieved based on the chromaticity values ​​obtained from the fluorescence intensity.

[0034] 4 μL of CNQDs@AuNPs@Asp probes were transferred and loaded onto a paper chip. 4 μL of Cys solution with concentrations of 220, 240, 260, 280, and 300 μmol / L were added sequentially. The results were recorded by taking pictures after 6 min. A Cys standard curve was established by linear fitting with Cys concentration as the x-axis and the corresponding RGB values ​​as the y-axis.

[0035] The following conclusions were drawn: when the Cys concentration is in the range of 220-300 μmol / L, the RGB values ​​show a good linear relationship with the Cys concentration. The standard curve is y = 0.1898x - 11.523, R0. 2 =0.9988. Different concentrations of Cys solutions were then measured, and their RGB values ​​were substituted into the standard curve to obtain the Cys concentration. The relative deviations were all within ±5%.

[0036] Please see Figures 1-5 This utility model provides an embodiment of a fluorescence quantitative analysis device, including a base 1; it also includes a light-shielding door 5 and a stage 6. An instrument light-shielding cover 2 and a handheld rechargeable ultraviolet lamp 3 are fixedly connected to the upper end of the base 1. An image acquisition cavity 4 is opened on the upper side of the front end of the instrument light-shielding cover 2. A light-shielding door 5 is provided at the front end of the image acquisition cavity 4. The stage 6 is slidably connected to the upper end of the inner wall of the image acquisition cavity 4. A positioning rivet 7 is provided at the upper end of the stage 6. A light-shielding plate 8 is fixedly connected to the front end of the stage 6. A camera body 9 is installed inside the image acquisition cavity 4. In use, the focal length of the camera body 9 is manually adjusted to clearly acquire the fluorescence image on the paper chip, improving detection accuracy. The handheld rechargeable ultraviolet lamp 3 is activated to provide a stable ultraviolet light source. An external light source is installed, and a light-shielding door 5 is installed. The accompanying software is opened, and the standard curve parameters, reaction time, and automatic recognition template are set. The positioning rivet 7 is selected according to the different shapes of the uric acid colorimetric paper chip. The positioning rivet 7 is installed on the stage 6. The replacement is flexible, which reduces the equipment cost and improves the versatility and practicality of the equipment. The uric acid colorimetric paper chip is placed on the positioning rivet 7, and a fluorescent sample is added. The start detection button is clicked on the experiment page. The stage 6 is slid into the image acquisition cavity 4 through the light-shielding door 5. The light shield 8 helps to form a sealed light-shielding environment to avoid interference from external light sources on the image. The camera body 9 automatically acquires the fluorescence image after the fluorescent material on the chip is excited and transmits the data. The final color value and concentration will be recorded in the software's historical data module.

[0037] Please see Figure 1 , Figure 3 and Figure 4In this embodiment, the instrument light shield 2, light shield door 5, stage 6, positioning rivet 7, and light shield plate 8 are all made of black resin material designed by 3D modeling, forming a sealed light-shielding environment to avoid interference from external light sources on the image and ensure the accuracy and stability of fluorescence detection. The rear end of the instrument light shield 2 is provided with a UV lamp inlet 10, which is compatible with the front end of the handheld rechargeable UV lamp 3. The right end of the handheld rechargeable UV lamp 3 is fixedly connected to a cable interface 11. The handheld rechargeable UV lamp 3 is inserted into the UV lamp inlet 10 to ensure that the UV lamp is installed firmly and can work normally to excite the fluorescent material on the paper chip. The cable interface 11 is used to transmit image data collected by the camera. The surface of the light shield door 5 is provided with a mounting hole 12 and an observation window 13. The rear end of the light shield door 5 is slidably connected to a light shield window 14, which is compatible with the observation window 13. The observation window 13 facilitates quick viewing of the sample status. Sliding the light shield window 14 restores the light-shielding environment and ensures that the fluorescence image acquisition is not interfered with by external light.

[0038] Please see Figure 2 , Figure 4 and Figure 5 In this embodiment, a limiting seat 15 is fixedly connected to the upper end of the inner wall of the image acquisition cavity 4. The two ends of the stage 6 are slidably connected along the limiting seat 15 and the mounting hole 12. The light shield 8 is larger than the mounting hole 12. The stage 6 is slid into the limiting seat 15 in the image acquisition cavity 4 through the mounting hole 12 to achieve positioning and fixation. The camera body 9 is a 4K macro camera. The lower end of the camera body 9 is fixedly connected to a lens 16 and a supplementary light 17. The lens 16 and the positioning rivet 7 are located on the same axis. The lens 16 provides high-quality image data for accurate fluorescence quantitative analysis. According to the needs of establishing a standard curve, the intensity of the supplementary light 17 is selected, or the supplementary light 17 is not used, and only the ultraviolet lamp excites the fluorescent material to emit fluorescence. The right end of the camera body 9 is threadedly connected to a nut 18. The other end of the nut 18 extends to the outside of the instrument light shield 2 and is threadedly connected to a nut 19. The camera body 9 is fixed in the image acquisition cavity 4 by the nut 18 and the nut 19, and the assembly is completed.

[0039] During operation, manually adjust the focal length of the camera body 9 to clearly acquire the fluorescence image on the paper chip. Turn on the handheld rechargeable UV lamp 3, install the light shield 5, open the accompanying software, set the standard curve parameters, reaction time, and automatic recognition template, select the positioning rivet 7 according to the different shapes of the uric acid colorimetric paper chip, install the positioning rivet 7 on the stage 6, place the uric acid colorimetric paper chip on the positioning rivet 7, and add the fluorescent sample. Click the start detection button on the experiment page, slide the stage 6 into the limiting seat 15 in the image acquisition cavity 4 through the mounting hole 12, and the light shield 8 helps to form a sealed light-shielding environment. The camera body 9 automatically acquires the fluorescence image after the fluorescent substance on the chip is excited and transmits the data. The final color value and concentration will be recorded in the software's historical data module.

[0040] Through the above steps, positioning rivets 7 are selected and installed on the stage 6 according to the different shapes of uric acid colorimetric paper chips. This allows for flexible replacement, reduces equipment costs, and improves the versatility and practicality of the equipment. The light-shielding door 5 and the light-shielding plate 8 help to form a sealed light-shielding environment, avoiding interference from external light sources on the image and ensuring the accuracy and stability of fluorescence detection. This solves the problems of insufficient sample adaptability and poor light-shielding effect of existing fluorescence quantitative analysis equipment, which affect the accuracy of fluorescence quantitative analysis.

Claims

1. A fluorescence quantitative analysis device, comprising a base (1); characterized in that: It also includes a light-shielding door (5) and a stage (6). The upper end of the base (1) is fixedly connected to an instrument light shield (2) and a handheld rechargeable ultraviolet lamp (3). An image acquisition cavity (4) is opened on the upper side of the front end of the instrument light shield (2). A light-shielding door (5) is set at the front end of the image acquisition cavity (4). A stage (6) is slidably connected to the upper end of the inner wall of the image acquisition cavity (4). A positioning rivet (7) is set at the upper end of the stage (6). A light shield (8) is fixedly connected to the front end of the stage (6). A camera body (9) is set inside the image acquisition cavity (4).

2. The fluorescence quantitative analysis device according to claim 1, characterized in that: The instrument light shield (2), light shield door (5), stage (6), positioning rivet (7) and light shield plate (8) are all made of black resin material designed by 3D modeling.

3. The fluorescence quantitative analysis device according to claim 1, characterized in that: The instrument's light shield (2) has a UV lamp inlet (10) at its rear end. The UV lamp inlet (10) is compatible with the front end of the handheld rechargeable UV lamp (3). The right end of the handheld rechargeable UV lamp (3) is fixedly connected to a cable interface (11).

4. The fluorescence quantitative analysis device according to claim 1, characterized in that: The surface of the light-proof door (5) is provided with mounting holes (12) and observation windows (13). The rear end of the light-proof door (5) is slidably connected to a light-shielding window (14), and the light-shielding window (14) and the observation window (13) are compatible.

5. The fluorescence quantitative analysis device according to claim 4, characterized in that: The upper end of the inner wall of the image acquisition cavity (4) is fixedly connected to the limiting seat (15), and the two ends of the stage (6) are slidably connected along the limiting seat (15) and the mounting hole (12). The light shield (8) is larger than the mounting hole (12).

6. The fluorescence quantitative analysis device according to claim 1, characterized in that: The camera body (9) is a 4K macro camera. The lower end of the camera body (9) is fixedly connected to a lens (16) and a fill light (17). The lens (16) and the positioning rivet (7) are located on the same axis.

7. The fluorescence quantitative analysis device according to claim 1, characterized in that: The right end of the camera body (9) is threaded with a nut (18), and the other end of the nut (18) extends to the outside of the instrument light shield (2) and is threaded with a nut (19).

Images