A μl sample-based point-of-care device and method

Abstract

The application discloses a kind of based on μL sample amount instant detection device and method, belong to in vitro diagnosis technical field.The device includes detection box, display screen, fluorescence detection module, acoustic flow control module and single board computer.Fluorescence detection module uses vertical coaxial light path, include camera, optical filter and light source, for high signal-to-noise ratio image capture;Acoustic flow control module includes acoustic flow control detection chip with sharp corner structure and excitation circuit, for generating acoustic flow vortex to accelerate trace sample mixing and reaction.Single board computer controls whole process and executes image analysis.The method is loaded by ≤10 μL sample, 1kHz~100kHz variable frequency sound field is applied to process 30s~600s, combined with coaxial light path acquisition and morphological algorithm, realize multi-index rapid quantitative detection.The application solves the problems of slow reaction, low signal-to-noise ratio and complex operation of trace sample in the prior art, with the advantages of small sampling amount, fast detection speed, high precision and portable intelligent, suitable for on-site instant detection.

Description

Technical Field

[0001] This invention belongs to the field of point-of-care testing (POCT) technology, specifically relating to a point-of-care testing device and method based on μL sample size. Background Technology

[0002] With the rapid development of in vitro diagnostic (IVD) technology, point-of-care testing (POCT) has been widely used in infectious disease screening, chronic disease management, and family health monitoring due to its advantages such as speed, portability, and on-site results. Existing POCT testing equipment is mainly based on technologies such as immunochromatographic test strips, acoustic fluidic detection chips, or small biochemical analyzers.

[0003] However, in practical applications, especially for rapid detection of trace samples, existing technologies still have the following significant technical problems and shortcomings: 1. Existing microfluidic or test strip-based detection devices primarily rely on capillary action or simple diffusion to mix samples and reagents. For whole blood samples with high viscosity, cell and serum separation is difficult, resulting in a slow reaction rate between the antigen / antibody and the label, often requiring 15-30 minutes or even longer to complete the reaction. Furthermore, due to the lack of an active mixing mechanism, micro-samples are prone to uneven reactions and large local concentration gradients, directly affecting the accuracy and repeatability of the test results.

[0004] 2. Traditional detection devices typically require sample volumes of tens to hundreds of microliters to ensure sufficient coverage of the detection area and obtain a valid signal. This is painful and invasive for infants, anemic patients, or patients requiring frequent monitoring, and makes it difficult to achieve truly real-time finger-prick blood testing. Current technologies often fail to detect micro- or even sub-micro-scale samples due to insufficient sample volume, resulting in incomplete channel filling or weak signals.

[0005] 3. Existing portable fluorescence detection devices often have relatively simple optical path designs, lacking precise coaxial calibration between the light source, filters, and detector, or employing a non-coaxial layout. This causes the excitation light to be easily scattered directly into the detector, generating strong background noise that masks weak fluorescence signals. Furthermore, existing devices have weak shielding capabilities against ambient light, resulting in large reading fluctuations and poor quantitative accuracy when detecting outdoors or in strong light environments, making it difficult to achieve laboratory-level detection sensitivity.

[0006] 4. Current trace detection equipment often has limited functionality, requiring sample pretreatment to be performed independently outside the device. This process is cumbersome and prone to errors due to human intervention. Furthermore, existing equipment offers limited result display methods, lacking intuitive guidance and intelligent result interpretation, making it difficult for ordinary users to independently complete the entire process from sampling to obtaining the report.

[0007] 5. In existing replaceable chip-based devices, the mechanical fixation methods between the chip and the device are often rudimentary, lacking precise positioning and locking mechanisms. During acoustic field vibration or movement, the chip is prone to slight displacement, leading to optical path focus shift or decreased acoustic field coupling efficiency. This results in poor consistency of test results between different batches or different devices, making it difficult to meet the stringent requirements of clinical quantification.

[0008] In summary, developing a point-of-care testing (POCT) device and method that can handle μL-level micro-samples, has active acoustic flow control for accelerated mixing, possesses a high signal-to-noise ratio coaxial optical path, is easy to operate, and produces accurate results is a pressing technical challenge in the current POCT field. Summary of the Invention

[0009] The technical problem to be solved by the present invention is to provide an instant detection device and method based on μL sample volume to address the shortcomings of the prior art, thereby solving the technical problems of high cost, poor portability and low concentration quantification efficiency of existing detection equipment, and realizing rapid, accurate and on-site quantitative detection of the concentration of the analyte.

[0010] The present invention adopts the following technical solution: A real-time detection device based on μL sample size, comprising: The detection box, top cover, and screen base together constitute the outer shell of the instant detection device. The display screen, fixed on the detection box, is used for device control and display of detection results; A detection compartment, disposed on the detection housing, is used to accommodate the acoustic fluid control detection chip; A fluorescence detection module, installed inside the housing, includes a camera, a filter, and a fluorescence light source. The fluorescence light source is used to excite the sample to emit fluorescence. The filter is disposed between the fluorescence light source and the camera to filter fluorescence signals. The camera is used to capture fluorescence images after the filter has been applied. The acoustic flow control module includes an acoustic flow control detection chip and an acoustic field excitation circuit. The acoustic flow control detection chip is detachably disposed in the detection chip compartment, and the acoustic field excitation circuit is used to generate an acoustic field signal to drive the acoustic flow control detection chip to work. The image analysis and processing module includes a single-board computer, which is electrically connected to the display screen, the camera, and the sound field excitation circuit. The single-board computer is used to control the operating parameters of the sound field excitation circuit, receive and process image data transmitted from the camera, and display the detection results on the display screen.

[0011] Preferably, the optical path of the fluorescence detection module is arranged vertically and coaxially, and the geometric centers of the camera, filter, fluorescence light source and acoustic fluid control detection chip are located on the same axis; The filter is fixed between the fluorescent light source and the camera, so that the fluorescence emitted by the acoustic fluid control detection chip excited by the fluorescent light source is filtered by the filter and enters the camera to obtain a fluorescence image of a single wavelength.

[0012] Preferably, the sound field excitation circuit includes a signal generator and a power amplifier; The single-board computer is electrically connected to the signal generator and is used to control the frequency, amplitude, and pulse waveform of the excitation signal; The signal generator is electrically connected to the power amplifier, and the power amplifier is electrically connected to the acoustic flow control detection chip, for amplifying the excitation signal and transmitting it to the acoustic flow control detection chip; The sound field frequency range generated by the sound field excitation circuit is 1kHz to 100kHz.

[0013] Preferably, the sound field excitation circuit further includes a shielded twisted pair cable; The single-board computer, signal generator, and power amplifier are connected via the shielded twisted-pair cable to reduce interference during high-frequency signal transmission. The power amplifier is connected to the piezoelectric layer beneath the acoustic flow control detection chip via a shielded twisted-pair cable, and is used to output pulse signals.

[0014] Preferably, the acoustic flow control detection chip has a multi-layer composite structure, including a flow channel layer, a reflective layer, and a piezoelectric layer; The flow channel layer is formed by bonding PDMS flow channels and glass cover glass, and the flow channel layer is tightly bonded to the reflective layer through silicone. The flow channel layer is provided with multiple branch flow channels, and sharp corner structures are arranged in the flow channel layer. The sharp corner structures are used to excite acoustic flow under the action of the sound field. The acoustic flow control detection chip is configured to simultaneously detect at least five different targets using a sample volume of up to 10 μL through the multiple branch channels.

[0015] Preferably, the reflective layer is made of a flexible lens, and the flow channel layer is made of PDMS flow channels and a glass cover glass through a plasma bonding process; The three layers—the flow channel layer, the reflective layer, and the piezoelectric layer—are tightly connected by adhesive bonding.

[0016] Preferably, it also includes a fixing clip and a fixing pin; After being fixed by the fixing clip, the acoustic flow control detection chip is placed in the detection chip compartment; The fixing clamp engages with the detection chip compartment via the fixing pin to restrict the positional movement of the acoustic flow control detection chip during the detection process.

[0017] Preferably, the detection box is made of opaque black resin SLA stereolithography 3D printing material; The detection box and the top cover are sealed together by a tenon and mortise structure; A light-shielding baffle is provided between the fluorescence detection module and the acoustic flow control module to prevent light crosstalk between the modules.

[0018] Preferably, the UI interface displayed on the screen includes an auxiliary guidance module, a device status module, a historical data module, an operation execution module, and a result presentation module; The operation execution module is equipped with a start measurement trigger control, which is used to trigger the single-board computer to control the sound field excitation circuit to work, and to display the vibration processing time progress and sample response process in real time. The result presentation module is used to display the corresponding images of multiple fluorescence detection areas in different regions after the detection is completed, and to display the measured fluorescence intensity values ​​of each target and the corresponding normal reference range. Based on the comparison results of the measured values ​​and the normal range, the module outputs the anomaly judgment results and suggestions.

[0019] Another technical solution of the present invention is a real-time detection method based on μL sample size, applied to the aforementioned real-time detection device and method based on μL sample size, comprising the following steps: S1. Add no more than 10 μL of the sample to be tested to the acoustic fluid control detection chip, fix it and place it in the detection chamber; use the coaxial optical path system to make the geometric center of the fluorescent light source, filter, camera and acoustic fluid control detection chip coincide, to ensure that the excitation light is incident perpendicularly and the fluorescence signal returns along the original path to be captured by the camera. S2. The control signal generator generates an excitation signal with a frequency of 1kHz~100kHz and an adjustable pulse waveform. After being amplified by the power amplifier, the signal is applied to the piezoelectric layer of the acoustic fluid control detection chip. Acoustic vortices are excited at the sharp corner structure of the flow channel layer and the vibration is continuously processed for 30s~600s to promote the full reaction between the serum and the detection area modified with the specific fluorescent probe. S3. After the reaction is complete, the fluorescent light source is controlled to emit excitation light of a specific wavelength. After the light passes through the sample, the background stray light is filtered out by the filter, and only the characteristic fluorescence of a single wavelength is allowed to pass through. The camera captures a high signal-to-noise ratio fluorescence image under the coaxial optical path. S4. The single-board computer performs morphological detection on the captured image, automatically segments the test areas corresponding to multiple branch channels, extracts the gray values ​​of each area and converts them into fluorescence intensity; combined with the pre-stored standard fluorescence intensity-substance concentration calibration curve, the concentration values ​​of each test substance are calculated. S5. Display the fluorescence images and corresponding measured concentration values ​​of each detection point in different areas on the display screen, and compare the measured values ​​with the normal reference range. If the values ​​exceed the range, automatically generate an abnormality judgment result and health guidance suggestions.

[0020] Compared with the prior art, the present invention has at least the following beneficial effects: A point-of-care testing device based on μL sample sizes integrates optical excitation and capture with active acoustic mixing within a portable housing, automating the process from sample input to result output. In particular, the collaborative design of the acoustic flow control module and fluorescence module solves the problems of slow reaction and uneven mixing of micro-samples under static conditions. A single-board computer acts as the central control unit, uniformly scheduling hardware resources, simplifying user operation, reducing human error, and significantly improving detection efficiency. This provides a solid hardware foundation for high-precision quantitative detection of μL-level samples, ensuring stable operation in non-laboratory environments such as homes and communities, and possesses significant clinical application value. Furthermore, the coaxial structure ensures that the excitation light is incident perpendicularly on the sample, and the generated fluorescence signal returns to the camera efficiently along the original path, minimizing optical path loss. Combined with the precise placement of filters between the light source and the camera, Rayleigh scattering and background stray light of the excitation light are effectively filtered out, allowing only characteristic fluorescence of specific wavelengths to pass through. This design overcomes the light crosstalk problem common in portable devices, enabling clear imaging even under weak fluorescence signals, significantly improving detection sensitivity and quantitative accuracy, and is particularly suitable for the early screening of low-concentration biomarkers.

[0021] Furthermore, the power amplifier linearly amplifies the weak signal, driving the piezoelectric layer to generate a high-intensity sound field, ensuring that energy is effectively transferred to the microchannels. The electronic control architecture avoids the limitations of fixed-frequency sound sources, enabling optimization of mixing efficiency for specific biomarkers, accelerating antigen-antibody binding reactions, and reducing detection time from tens of minutes to minutes, while ensuring the uniformity and repeatability of the reaction process.

[0022] Furthermore, acoustic flow control technology involves high-frequency, high-voltage signals, which are highly susceptible to electromagnetic radiation interference in sensitive image acquisition circuits. Shielded twisted-pair cabling effectively suppresses external noise intrusion and internal signal crosstalk during signal transmission, ensuring the purity of the excitation waveform. This not only protects the stable operation of the single-board computer and camera, preventing image noise or artifacts, but also guarantees the stability of the sound field output, avoiding a decrease in mixing efficiency due to signal fluctuations. This design enhances the reliability of the equipment in complex electromagnetic environments and ensures the consistency of detection data.

[0023] Furthermore, the sharp-angled structure generates strong acoustic vortices under the influence of the acoustic field, actively breaking through laminar flow limitations, rapidly separating blood cells, and promoting the mixing of serum and reagents, thus solving the problem of slow diffusion of micro-samples. The multi-branched flow channel design allows for the parallel processing of at least five different targets in a single test, significantly increasing throughput. Sample volumes of up to 10 μL greatly reduce patient discomfort during blood collection, making it particularly suitable for infants and patients with anemia. This microstructural design integrates complex laboratory pretreatment steps within the chip, which is key to achieving truly portable, high-throughput POCT.

[0024] Furthermore, the glass cover glass possesses excellent hydrophilicity and light transmittance. As a component of the flow channel layer, it effectively promotes rapid liquid flow to the detection area and enhances the collection efficiency of fluorescence signals while reducing background absorption. The reflective layer utilizes a flexible lens with adhesive backing, whose adhesive surface is directly bonded to the piezoelectric layer, achieving a tight interlayer bond. This avoids acoustic wave loss caused by interface mismatch in traditional multilayer bonding processes, ensuring efficient transmission of acoustic energy generated by the piezoelectric layer to the flow channel layer. The glass substrate is bonded to the reflective layer with silicone. The silicone layer provides robust encapsulation while buffering vibrations and adapting to differences in thermal stress between materials. This composite encapsulation method offers high reliability: the tight bonding between the piezoelectric layer and the reflective layer ensures stable energy transmission under high-pressure acoustic fields, while the silicone bonding between the reflective layer and the flow channel layer effectively prevents delamination and leakage. The overall structure maintains durability under repeated vibrations, extending the lifespan of consumables. Simultaneously, the silicone bonding process has lower requirements for surface flatness, reducing the risk of cost fluctuations in mass production.

[0025] Furthermore, the precise positioning of the chip in the X, Y, and Z dimensions is achieved through the cooperation of the fixing pin and the detection chip compartment, ensuring that the detection area accurately repositions itself at the optical path focal point and acoustic antinode position after each chip replacement. The preload provided by the fixing clamp prevents minute displacement or resonance noise of the chip during acoustic field vibration, avoiding optical path defocusing or decreased acoustic coupling efficiency due to positional misalignment. This reliable mechanical connection significantly reduces intra-batch and inter-batch variability, improves the overall measurement repeatability of the equipment, and meets the stringent requirements of clinical quantitative testing.

[0026] Furthermore, the black SLA material absorbs external stray light and internal reflected light at the source, while the tenon and mortise structure ensures the airtightness and light tightness of the casing, eliminating interference from ambient light leakage on weak fluorescence signals. Light-shielding baffles between modules further isolate the fluorescence excitation light from potential light pollution from the acoustic flow control circuitry. This comprehensive light-shielding design allows the device to operate in bright environments without relying on a large darkroom, greatly expanding its application scenarios.

[0027] Furthermore, the auxiliary guidance module reduces the professional requirements for operators through graphic demonstrations; the real-time progress bar allows users to clearly understand the reaction process, alleviating anxiety while waiting. Most importantly, the results presentation module not only displays the original images and values ​​but also automatically compares them to normal reference ranges, outputting anomaly assessments and health recommendations. This integrated output of "detection + diagnostic advice" compensates for the lack of experience among primary healthcare personnel, helping users quickly understand test results and take appropriate measures, significantly improving the user experience and practical application value of POCT equipment.

[0028] A real-time detection method based on μL sample size is proposed. Optical path calibration ensures the consistency of signal acquisition benchmark; frequency conversion acoustic field processing actively intervenes in reaction dynamics, transforming the natural diffusion process into forced convection mixing, significantly shortening the detection time; narrowband acquisition strategy combined with hardware filtering further purifies the signal; morphological segmentation algorithm automatically removes background interference and accurately extracts the grayscale value of the region of interest. This method not only solves the problems of difficulty, slowness, and inaccuracy in detecting micro-samples, but also eliminates human interference through standardized operating procedures, ensuring high sensitivity, high specificity, and high repeatability of the detection results.

[0029] In summary, this invention achieves rapid, accurate, and simultaneous multi-indicator detection of μL-level samples through integrated acoustic-optical-mechatronic technology. It addresses the slow response time issue by utilizing acoustic-fluidic active mixing, ensures a high signal-to-noise ratio through coaxial optical paths and anti-interference design, and simplifies operation through intelligent algorithms and user-friendly UI interaction. The overall solution significantly reduces blood collection volume and testing time, improves the sensitivity, repeatability, and user experience of POCT equipment, and possesses extremely high clinical translational value.

[0030] The technical solution of the present invention will be further described in detail below with reference to the accompanying drawings and embodiments. Attached Figure Description

[0031] To more clearly illustrate the technical solutions in the embodiments of the present invention, the accompanying drawings used in the following description of the relative embodiments will be briefly introduced. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0032] Figure 1 This is an overall diagram of the detection device of the present invention. Figure 2 Exploded view of the detection device of the present invention Figure 3 This is an overall diagram of the detection compartment of the present invention. Figure 4 This is an unfolded diagram of the detection compartment of the present invention. Figure 5 Exploded view of the detection piece of this invention Figure 6 This is the start UI interface of the detection device of the present invention. Figure 7 This is the UI interface for displaying the detection results of the present invention.

[0033] The components include: 1. Detection box; 2. Display screen; 3. Top cover; 4. Screen base; 5. Single-board computer; 6. Signal generator; 7. Switching power supply; 8. Power amplifier; 9. Camera; 10. Filter; 11. Fluorescent light source; 12. Detection chip compartment; 15. Acoustic flow control detection chip; 16. Fixing clip; 17. Fixing pin; 18. Flow channel layer; 19. Reflective layer; 20. Piezoelectric layer. Detailed Implementation

[0034] 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, not all, of the embodiments of the present invention. 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.

[0035] In the description of this invention, it should be understood that the terms "center," "longitudinal," "lateral," "upper," "lower," "front," "rear," "left," "right," "vertical," "horizontal," "top," "bottom," "inner," "outer," "one side," "one end," and "one side," etc., indicating orientation or positional relationships based on the orientation or positional relationships shown in the accompanying drawings, are only for the convenience of describing the invention and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation, and therefore should not be construed as a limitation of the invention. Furthermore, the terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance or implicitly specifying the number of indicated technical features. Thus, a feature defined with "first" or "second" may explicitly or implicitly include one or more of that feature. In the description of this invention, unless otherwise stated, "a plurality of" means two or more.

[0036] In the description of this invention, it should be noted that, unless otherwise explicitly specified and limited, the terms "installation," "connection," and "linking" should be interpreted broadly. For example, they can refer to a fixed connection, a detachable connection, or an integral connection; they can refer to a mechanical connection or an electrical connection; they can refer to a direct connection or an indirect connection through an intermediate medium; and they can refer to the internal connection of two components. Those skilled in the art can understand the specific meaning of the above terms in this invention based on the specific circumstances.

[0037] It should be understood that, when used in this specification and the appended claims, the terms "comprising" and "including" indicate the presence of the described features, integrals, steps, operations, elements and / or components, but do not exclude the presence or addition of one or more other features, integrals, steps, operations, elements, components and / or collections thereof.

[0038] It should also be understood that the terminology used in this specification is for the purpose of describing particular embodiments only and is not intended to limit the invention. As used in this specification and the appended claims, the singular forms “a,” “an,” and “the” are intended to include the plural forms unless the context clearly indicates otherwise.

[0039] It should also be further understood that the term "and / or" as used in this specification and the appended claims refers to any combination of one or more of the associated listed items and all possible combinations, and includes such combinations.

[0040] The accompanying drawings illustrate various structural schematic diagrams according to embodiments disclosed in this invention. These drawings are not to scale, and some details have been enlarged for clarity, and some details may have been omitted. The shapes of the various regions and layers shown in the drawings, as well as their relative sizes and positional relationships, are merely exemplary and may deviate from reality due to manufacturing tolerances or technical limitations. Furthermore, those skilled in the art can design regions / layers with different shapes, sizes, and relative positions as needed.

[0041] This invention provides a point-of-care testing device and method based on μL sample volume. Utilizing acoustic fluid control technology, it shortens the sample processing flow and accelerates the reaction time between the sample and the detection reagent. It offers advantages such as low detection cost, high sensitivity, short detection time, easy integration, and small device size. It is suitable for various point-of-care testing scenarios, including early medical diagnosis in hospitals and homes, as well as field applications, and has significant practical application value.

[0042] Please see Figure 1The present invention discloses an instantaneous detection device based on μL sample size, comprising a detection box 1, a display screen 2, a top cover 3, a screen base 4, a single-board computer 5, a signal generator 6, a switching power supply 7, a power amplifier 8, a camera 9, a filter 10, a fluorescent light source 11, a detection chamber 12, an acoustic flow control detection chip 15, a fixing clip 16, a fixing pin 17, a flow channel layer 18, a reflective layer 19, and a piezoelectric layer 20.

[0043] The display screen 2 is fixed on the detection box 1 and located above the device. It is used to display the test results, analysis data and operation interface. The detection box 1 can be made of opaque black resin SLA stereolithography 3D printing material, which reduces product cost and equipment weight, and provides the light-proof environment required for fluorescence detection. The detection chip compartment 12 is located next to the display screen 2 and uses a tenon and mortise structure to realize the switch, which facilitates the insertion and removal of the acoustic flow control detection chip 15. The display screen 2 provides a visual display of the sample test results.

[0044] The detection box 1 is made by 3D printing and uses black ABS or PLA material to balance structural stability and lightweight requirements. The top cover of the main body of the detection box 1 is sealed with a tenon and mortise structure to ensure the airtightness of the detection environment. A light-shielding baffle is set between the fluorescence detection module and the acoustic flow control module to effectively avoid light crosstalk between modules and provide a stable light-shielding environment for fluorescence detection.

[0045] Display screen 2 is used for device control and test result display. Its UI interface is configured with an auxiliary guidance module, a device status module, a historical data module, an operation execution module, and a result presentation module. The auxiliary guidance module provides illustrated demonstrations of the operation steps for the built-in detection device, allowing users to access and review them at any time during operation, thus lowering the barrier to entry for operation. The device status module is used to display the operating parameters (such as sound field frequency, excitation signal amplitude, etc.) and hardware conditions (such as voltage and current of each component) of the detection device in real time, so as to facilitate users to monitor the operating status of the device. The historical data module is used to store and retrieve historical measurement records. It allows users to query the results data of the corresponding test batches according to the time dimension, which facilitates data traceability and analysis. The operation execution module is used to set the start measurement trigger control. After triggering, the current measurement process can be displayed in real time, including the vibration treatment time progress and the sample reaction process, so that users can intuitively understand the detection progress. The results presentation module is used to display the corresponding images of the five fluorescence detection areas in the results interface after the detection is completed. It also displays the measured fluorescence intensity values ​​of each target and the corresponding normal reference range. Based on the comparison between the measured values ​​and the normal range, it outputs the anomaly judgment result and matches the corresponding suggestion information, including health guidance or suggestions for further testing.

[0046] Please see Figure 2 The camera 9, filter 10, fluorescence light source 11, and detection plate chamber 12 of the fluorescence detection module are arranged in a vertical optical path. The optical path design is simple and safe, and facilitates light shielding and the capture of fluorescence images by the camera 9. The single-board computer 5, signal generator 6, and power amplifier 8 are mounted at the front right end of the device. This layout is conducive to signal transmission and reduces external interference to high-frequency signals. At the same time, the fluorescence detection module and the acoustic flow control module are separated by a light shield to avoid light crosstalk. The light source excites the detection plate to emit fluorescence, and a single fluorescence image for analysis is obtained through the filter 10. After obtaining the sample fluorescence intensity, the single-board computer 5 directly visualizes the detection results according to the pre-calibrated curve.

[0047] All components of the fluorescence detection module are fixed within the detection device housing using an interference fit, ensuring that the geometric centers of the lens, fluorescence light source, and acoustic fluid control chip are on the same axis, thus guaranteeing optical path stability. During operation, the fluorescence light source excites the analyte on the acoustic fluid control chip 15 to react with the reagent and emit fluorescence. The fluorescence is filtered by a light filter to obtain a single fluorescence image for analysis, avoiding stray light interference with the detection results.

[0048] The acoustic flow control module includes a sound field excitation circuit and an acoustic flow control detection chip 15. The sound field excitation circuit comprises a single-board computer 5, a signal generator 6, and a power amplifier 8. These components are connected via shielded twisted-pair cables to reduce interference during high-frequency signal transmission. The single-board computer 5 controls the frequency, amplitude, pulse waveform, and other parameters of the excitation signal. The sound field frequency ranges from 1kHz to 100kHz, meeting the requirements of acoustic flow control technology. The sound field excitation circuits are interconnected via shielded twisted-pair cables.

[0049] Please see Figure 3 and Figure 4 After the acoustic flow control detection chip 15 is fixed by the fixing clip 16, it is placed in the detection chip compartment 12. This design facilitates the addition of sample reagents to the acoustic flow control detection chip 15 and ensures the stability of the chip position during the imaging process. The power amplifier 8 is connected to the bottom of the acoustic flow control detection chip 15 through a shielded twisted pair cable to output pulse signals, which are converted into the sound field required for operation through the piezoelectric layer 20.

[0050] The acoustic flow control detection chip 15 has multiple branch channels, enabling the detection of at least five different targets using a sample of up to 10 μL. The number of targets detected depends on the number of branch channels on the acoustic flow control detection chip 15.

[0051] Please see Figure 5 The acoustic flow control detection chip 15 consists of a flow channel layer 18, a reflective layer 19, and a piezoelectric layer 20. The three layers are tightly connected by plasma bonding and adhesive bonding processes, resulting in a strong connection and good sealing. The flow channel layer 18 has sharp-angled structures arranged inside, which are used to excite acoustic flow under the action of an acoustic field, accelerating the reaction between the sample and the reagent. The reflective layer 19 is a flexible lens, bonded between the piezoelectric layer 20 and the flow channel layer 18, which can enhance the fluorescence reflection effect and improve the detection signal intensity. Optionally, the bonding between the layers of the acoustic flow control detection chip 15 can be selected by ion bonding or adhesive bonding.

[0052] Optionally, depending on the number and type of the substance being detected, different acoustic fluid control detection chips can be used to adapt to the detection requirements.

[0053] A real-time detection method based on μL sample size includes the following steps: S1. Add the sample to be tested into the acoustic flow control detection chip 15, fix it with the fixing clip 16, and then place it into the detection chip compartment 12; S2. Start the program in the operation interface of the display screen 2, and use the single-board computer 5, signal generator 6 and power amplifier 8 to generate vibrations for 30s~600s to process the acoustic flow control detection chip 15. S3. After the sample reaction in the acoustic fluid control detection chip 15 is completed, the camera 9 captures the fluorescence image after filtering by the filter. After the camera captures a fluorescence image, it transmits the image data to a single-board computer 5. The single-board computer 5 first performs morphological detection on the image, segmenting the region to be tested using morphological algorithms. Then, it calculates the fluorescence intensity of the region based on the image's grayscale values. Combined with a pre-calibrated standard fluorescence intensity-substance concentration curve, it completes the quantitative analysis of the concentration of the analyte and displays the detection results and related analysis content on the display screen 2. Optionally, the single-board computer 5 allows the integration of more complex machine learning algorithms to further improve the accuracy and timeliness of the detection results.

[0054] S4, the single-board computer 5, based on the obtained image data, performs image processing and recognition analysis to obtain the fluorescence intensity of each marker on the acoustic fluid control detection chip 15; S5. The single-board computer 5 visualizes the obtained fluorescence intensity data on the UI interface of the display screen 2.

[0055] Please see Figure 6The start screen includes a configuration module for auxiliary guidance, a device status module, a historical data module, an operation execution module, and a result presentation module.

[0056] Please see Figure 7 The results display interface shows the corresponding images of the five fluorescence detection areas in separate regions. It also displays the measured fluorescence intensity values ​​of each target and the corresponding normal reference range. Based on the comparison between the measured values ​​and the normal range, the results presentation module outputs the anomaly judgment result and matches the corresponding suggestion information, which includes health guidance or suggestions for further testing.

[0057] This invention features an integrated design that keeps the device weight under 2kg, allowing it to be carried by a single person to the field (such as patient wards, homes, water quality monitoring points, and food testing sites) without relying on large laboratory environments. The entire detection process is automated, requiring no manual intervention in sample processing and data calculation. The detection time for a single sample is reduced to less than 10 minutes, with minimal concentration quantification error, meeting the demands for rapid and accurate detection. Core components utilize common devices such as ultraviolet lamps and small cameras, resulting in a manufacturing cost that is only 1 / 10 of traditional mass spectrometry equipment, facilitating large-scale production and widespread adoption. By changing the filters to different wavelengths, it can adapt to the detection needs of various substances, covering multiple fields such as environmental monitoring, food testing, and medical diagnosis.

[0058] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, 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, not all, of the embodiments of the present invention. The components of the embodiments of the present invention described and shown in the accompanying drawings can generally be arranged and designed in various different configurations. Therefore, the following detailed description of the embodiments of the present invention provided in the accompanying drawings is not intended to limit the scope of the claimed invention, but merely to illustrate selected embodiments of the invention. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without inventive effort are within the scope of protection of the present invention.

[0059] Example 1 This embodiment uses whole blood samples from clinical patients with acute myocardial infarction as the test samples to detect the concentrations of five diagnostic markers for acute myocardial infarction: CK-MB, cTnI, Myo, CRP, and NT-proBNP. The specific detection procedure is as follows: Sample preparation and loading: Take 10 μl of patient whole blood sample and slowly drip it into the sample inlet of the acoustic flow control detection chip 15. The sample will automatically fill into each branch channel under capillary action. Use the fixing clip 16 to fix the acoustic flow control detection chip 15 after loading the sample firmly to avoid chip displacement during the detection process. Then put it into the detection chip compartment 12 and close the compartment door with the tenon and mortise structure.

[0060] Sound field processing and reaction acceleration: In the operation interface of the display screen 2, after confirming that the operation steps are correct by checking the auxiliary guidance module, click the "Start Measurement" trigger control of the operation execution module to start the detection program; the single-board computer 5 issues a control command, the signal generator 6 generates an excitation signal with set parameters (frequency, amplitude, pulse waveform), which is amplified by the power amplifier 8 and transmitted to the piezoelectric layer 20 of the acoustic flow control detection chip 15 through a shielded twisted pair cable to generate a sound field with a frequency of 1kHz~100kHz; the sound field excites acoustic flow at the sharp corner structure of the flow channel layer 18, forming eddies to capture blood cells, causing the serum to flow to the detection area modified with specific fluorescent probes in each branch flow channel, and the five markers specifically bind to the corresponding fluorescent probes respectively; during this process, the operation execution module displays the vibration processing time progress in real time. In this embodiment, the vibration processing time is set to 10 minutes to ensure that the reaction is fully carried out.

[0061] Fluorescence image capture: After the sample reaction in the acoustic fluid control detection chip 15 is completed, the single-board computer 5 automatically triggers the fluorescence detection module to work. The fluorescence light source 11 emits excitation light, which illuminates the detection area to excite fluorescence. After the fluorescence is filtered by the filter 10, a single wavelength fluorescence signal is obtained. The camera 9 starts shooting, capturing the fluorescence image of each detection area, and transmitting the image data to the single-board computer 5 in real time.

[0062] Image analysis and concentration calculation: After receiving fluorescence image data, the single-board computer 5 first segments 5 test areas using morphological algorithms, and then calculates the fluorescence intensity based on the image grayscale value of each area; combined with the pre-calibrated fluorescence intensity-marker content standard curve, the concentration values ​​of each marker are calculated through the built-in algorithm; the device status module displays the data analysis process in real time to ensure the stability of the calculation process.

[0063] Results Display and Output: After data analysis is completed, the results presentation module on screen 2 displays the corresponding images of the five fluorescence detection areas in separate regions. It also displays the measured fluorescence intensity values ​​of five biomarkers—CK-MB, cTnI, Myo, CRP, and NT-proBNP—along with their corresponding normal reference ranges. Based on the comparison between the measured values ​​and the normal ranges, the system automatically outputs anomaly judgment results. If the concentration of a biomarker exceeds the normal range, it will match the corresponding health guidance or suggestions for further testing. Users can store the test results through the historical data module for easy reference in subsequent follow-up visits and re-examinations.

[0064] Comparison of Implementation Results In this embodiment, the detection device of the present invention reduces the actual operation time required for traditional ELISA testing from 2 hours to 15 minutes in the determination of whole blood samples from patients with acute myocardial infarction. The liquid handling steps are simplified from 5 transfer steps to a single step, significantly improving detection efficiency. Furthermore, the detection results of the present invention for the five biomarkers show good consistency with those of traditional ELISA testing, with small concentration quantification errors, meeting the accuracy requirements of clinical testing. Specific comparison results are shown in Table 1.

[0065] Example 2 This embodiment uses five environmental monitoring indicators (lead, mercury, cadmium, chromium, and arsenic) in drinking water as the test samples. The trace concentration is detected using the real-time detection device based on μL sample volume of this invention. The specific detection process is as follows: Sample preparation and loading: Take 8 μL of drinking water sample to be tested and slowly add it dropwise to the sample inlet of the acoustic flow control detection chip 15 adapted for heavy metal detection. The sample will automatically fill the five branch channels of the chip under capillary action. Use the fixing clip 16 to fix the chip after loading the sample firmly to prevent the chip from shifting during the detection process. Then put it into the detection chip compartment 12 and close the compartment door with the tenon and mortise structure to ensure that the chip is in the optimal position for the optical path and sound field.

[0066] Sound field processing and reaction acceleration: In the operation interface of the display screen 2, after confirming that the operation steps for heavy metal detection are correct by referring to the auxiliary guidance module, click the "Start Measurement" trigger control of the operation execution module to start the detection program; the single-board computer 5 issues a control command, the signal generator 6 generates a 50kHz sine wave excitation signal, which is amplified by the power amplifier 8 and transmitted to the piezoelectric layer 20 of the acoustic flow control detection chip 15 through a shielded twisted pair cable to generate a stable sound field; the sound field excites acoustic flow vortices at the sharp corner structure of the flow channel layer 18, and the vibration processing is carried out for 60s, which promotes the full binding of the drinking water sample with the heavy metal-specific fluorescent probe modified in the flow channel. The operation execution module displays the vibration processing time progress in real time.

[0067] Fluorescence image capture: After the sample and probe reaction is completed, the single-board computer 5 automatically triggers the fluorescence detection module to work. The fluorescence light source 11 emits excitation light of a specific wavelength, which illuminates the detection area of ​​the chip to excite fluorescence. After the background stray light is filtered out by the filter 10, only the characteristic fluorescence of a single wavelength is allowed to pass through. The camera 9 captures high signal-to-noise ratio fluorescence images of each detection area under the coaxial optical path and transmits the image data to the single-board computer 5 in real time.

[0068] Image analysis and concentration calculation: After receiving fluorescence image data, the single-board computer 5 automatically segments the test areas corresponding to the five branch channels through morphological algorithms, extracts the gray values ​​of each area and converts them into fluorescence intensity; combined with the pre-stored standard calibration curve of fluorescence intensity-concentration of heavy metal ions, the actual concentration values ​​of five heavy metal ions, namely lead, mercury, cadmium, chromium and arsenic, are calculated by the built-in algorithm. The device status module displays the data analysis process in real time to ensure the stability of the calculation process.

[0069] Results Display and Output: After data analysis is completed, the results presentation module on screen 2 displays the corresponding images of the five fluorescence detection areas in separate regions. It also displays the measured fluorescence intensity values, measured concentration values, and normal reference ranges in the national drinking water standards for the five heavy metal ions. Based on the comparison results between the measured concentrations and the reference ranges, the system automatically outputs anomaly judgment results. If the concentration of a certain heavy metal ion exceeds the standard, the system immediately matches the corresponding water quality treatment recommendations. The test results can be stored through the historical data module, which facilitates subsequent data traceability and trend analysis of water quality monitoring.

[0070] The detection process in this embodiment is fully automated, taking only 8 minutes from sample loading to result output. Compared with traditional atomic absorption spectrometry, it does not require professional laboratory equipment and operators, and can be completed on-site at drinking water sources, homes, etc. Moreover, the relative error between the detection results and the atomic absorption spectrometry results is within 5%, meeting the accuracy requirements of rapid on-site environmental detection. This fully demonstrates the versatility and practicality of the device in the field of environmental monitoring.

[0071] The detection device of this invention requires no professional operators or large laboratory equipment. It can be carried to wards, homes and other field scenarios for use. It has low detection cost and strong versatility. It is not only suitable for the detection of acute myocardial infarction markers, but can also be adapted to the detection of trace substance concentrations in environmental monitoring, food testing and other fields by changing the filter and detection chip. It has important practical application value.

[0072] In summary, this invention provides a point-of-care testing device and method based on μL sample volume, requiring only a maximum of 10μL of sample to simultaneously detect at least five indicators, greatly reducing patient discomfort and making it particularly suitable for infants and children and for micro-volume blood collection scenarios. Acoustic flow control technology shortens reaction time to the minute level and improves mixing efficiency several times over. The vertical coaxial optical path and narrow-band filter design significantly improve the signal-to-noise ratio. Combined with morphological image analysis algorithms, it achieves high-precision quantification of low-concentration biomarkers, with detection sensitivity reaching laboratory levels. The integrated design eliminates reliance on large peripherals, and the fully enclosed light-shielding structure adapts to various environments. Intelligent UI guidance and automatic interpretation functions enable non-professionals to independently complete high-quality testing, promoting the widespread application of POCT in primary care and home settings.

[0073] The above content is only for illustrating the technical concept of the present invention and should not be construed as limiting the scope of protection of the present invention. Any modifications made to the technical solution based on the technical concept proposed in this invention shall fall within the scope of protection of the claims of this invention.

Claims

1. A real-time detection device based on μL sample size, characterized in that, include: The detection box (1), top cover (3) and screen base (4) together constitute the outer shell of the instant detection device; The display screen (2) is fixed on the detection box (1) and is used for device control and display of detection results; The detection compartment (12) is disposed on the detection box (1) and is used to accommodate the acoustic flow control detection chip (15). The fluorescence detection module, installed inside the housing, includes a camera (9), a filter (10), and a fluorescence light source (11). The fluorescence light source (11) is used to excite the sample to emit fluorescence. The filter (10) is placed between the fluorescence light source (11) and the camera (9) to filter fluorescence signals. The camera (9) is used to capture the fluorescence image after passing through the filter (10). The acoustic flow control module includes an acoustic flow control detection chip (15) and an acoustic field excitation circuit. The acoustic flow control detection chip (15) is detachably disposed in the detection chip compartment (12). The acoustic field excitation circuit is used to generate an acoustic field signal to drive the acoustic flow control detection chip (15) to work. The image analysis and processing module includes a single-board computer (5), which is electrically connected to the display screen (2), the camera (9) and the sound field excitation circuit, respectively. It is used to control the working parameters of the sound field excitation circuit, receive image data transmitted by the camera (9) and process and analyze it, and display the detection results on the display screen (2).

2. The real-time detection device based on μL sample size according to claim 1, characterized in that, The optical path of the fluorescence detection module is arranged vertically and coaxially. The geometric centers of the camera (9), filter (10), fluorescence light source (11) and acoustic flow control detection chip (15) are located on the same axis. The filter (10) is fixed between the fluorescent light source (11) and the camera (9), so that the fluorescence emitted by the acoustic fluid control detection chip (15) excited by the fluorescent light source (11) is filtered by the filter (10) and enters the camera (9) to obtain a fluorescence image of a single wavelength.

3. The real-time detection device based on μL sample size according to claim 1, characterized in that, The sound field excitation circuit includes a signal generator (6) and a power amplifier (8). The single-board computer (5) is electrically connected to the signal generator (6) and is used to control the frequency, amplitude and pulse waveform of the excitation signal; The signal generator (6) is electrically connected to the power amplifier (8), and the power amplifier (8) is electrically connected to the acoustic flow control detection chip (15) to amplify the excitation signal and transmit it to the acoustic flow control detection chip (15). The sound field frequency range generated by the sound field excitation circuit is 1kHz to 100kHz.

4. The real-time detection device based on μL sample size according to claim 3, characterized in that, The sound field excitation circuit also includes a shielded twisted pair cable (14). The single-board computer (5), signal generator (6) and power amplifier (8) are connected by the shielded twisted pair cable (14) to reduce interference during high-frequency signal transmission; The power amplifier (8) is connected to the piezoelectric layer (20) below the acoustic flow control detection chip (15) via a shielded twisted pair cable (14) for outputting pulse signals.

5. The real-time detection device based on μL sample size according to claim 1, characterized in that, The acoustic flow control detection chip (15) has a multi-layer composite structure, including a flow channel layer (18), a reflective layer (19), and a piezoelectric layer (20). The reflective layer (19) is bonded between the flow channel layer (18) and the piezoelectric layer (20); The flow channel layer (18) is provided with multiple branch flow channels, and the flow channel layer (18) is arranged with sharp corner structures, which are used to excite acoustic flow under the action of the sound field; The acoustic flow control detection chip (15) is configured to simultaneously detect at least five different targets using the multiple branch channels with a sample volume of up to 10 μL.

6. The real-time detection device based on μL sample size according to claim 5, characterized in that, The reflective layer (19) is made of a flexible lens, and the flow channel layer (18) is made of PDMS flow channel and glass cover glass through plasma bonding process; The three layers, namely the flow channel layer (18), the reflective layer (19) and the piezoelectric layer (20), are tightly connected by adhesive bonding.

7. The real-time detection device based on μL sample size according to claim 1, characterized in that, It also includes a retaining clip (16) and a retaining pin (17); The acoustic flow control detection chip (15) is fixed by the fixing clip (16) and placed in the detection chip compartment (12); The fixing clip (16) cooperates with the detection chip compartment (12) through the fixing pin (17) to restrict the position movement of the acoustic flow control detection chip (15) during the detection process.

8. The real-time detection device based on μL sample size according to claim 1, characterized in that, The detection box (1) is made of opaque black resin SLA stereolithography 3D printing material; The detection box (1) and the top cover (3) are sealed together by a tenon and mortise structure; A light-shielding baffle is provided between the fluorescence detection module and the acoustic flow control module to prevent light crosstalk between the modules.

9. The real-time detection device based on μL sample size according to claim 1, characterized in that, The UI interface displayed on the display screen (2) includes an auxiliary guidance module, a device status module, a historical data module, an operation execution module, and a result presentation module; The operation execution module is equipped with a start measurement trigger control, which is used to trigger the single-board computer (5) to control the sound field excitation circuit to work, and to present the vibration processing time progress and sample reaction process in real time; The result presentation module is used to display the corresponding images of multiple fluorescence detection areas in different regions after the detection is completed, and to display the measured fluorescence intensity values ​​of each target and the corresponding normal reference range. Based on the comparison results of the measured values ​​and the normal range, the module outputs the anomaly judgment results and suggestions.

10. A real-time detection method based on μL sample size, applied to the real-time detection apparatus and method based on μL sample size as described in any one of claims 1 to 9, characterized in that, Includes the following steps: S1. Add no more than 10 μL of the sample to be tested to the acoustic fluid control detection chip (15), fix it and put it into the detection chamber (12); use the coaxial optical path system to make the geometric center of the fluorescent light source (11), filter (10), camera (9) coincide with the acoustic fluid control detection chip (15) to ensure that the excitation light is incident perpendicularly and the fluorescence signal returns along the original path to be captured by the camera (9); S2. The control signal generator (6) generates an excitation signal with a frequency of 1kHz~100kHz and an adjustable pulse waveform. After being amplified by the power amplifier (8), the signal is applied to the piezoelectric layer (20) of the acoustic flow control detection chip (15). Acoustic flow vortices are excited at the sharp corner structure of the flow channel layer (18), and the vibration is continuously processed for 30s~600s to promote the serum to fully react with the detection area modified with the specific fluorescent probe. S3. After the reaction is completed, control the fluorescent light source (11) to emit excitation light of a specific wavelength. After the light passes through the sample, it is filtered by the filter (10) to remove background stray light and only allow the characteristic fluorescence of a single wavelength to pass through. The camera (9) captures a high signal-to-noise ratio fluorescence image under the coaxial optical path. S4. The single-board computer (5) performs morphological detection on the captured image, automatically segments the test areas corresponding to multiple branch channels, extracts the gray values ​​of each area and converts them into fluorescence intensity; and calculates the concentration values ​​of each test substance by combining the pre-stored standard fluorescence intensity-substance concentration calibration curve. S5. Display the fluorescence images of each detection point and the corresponding measured concentration values ​​in different areas on the display screen (2), and compare the measured values ​​with the normal reference range. If the values ​​exceed the range, automatically generate abnormal judgment results and health guidance suggestions.

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