A method, apparatus, equipment and medium for flow-directed hybridization analysis

Abstract

This invention relates to the field of bioengineering technology and discloses a flow-through hybridization analysis method, apparatus, device, and medium. The method includes inkjet-spraying a paper-based chip to generate a circular spot array; wherein each circular spot is composed of several microdroplets; calculating the space-filling coefficient of the microdroplets; if the space-filling coefficient is less than a set threshold, optimizing the space-filling coefficient until it is not less than the threshold, resulting in a paper-based chip with uniformly distributed microdroplets; performing a flow-through hybridization reaction on the paper-based chip with uniformly distributed microdroplets to obtain a paper-based chip carrying a hybridization signal; and performing qualitative and quantitative detection and analysis on the paper-based chip carrying the hybridization signal to obtain the analysis results. This invention improves the detection accuracy of flow-through hybridization analysis and makes the signal distribution of the circular spots more regular, enhancing the readability of the results.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of bioengineering, and more particularly, to a flow-through hybridization analysis method, device, equipment and medium. BACKGROUND

[0002] In the field of biomedical detection, the flow-through hybridization analysis platform is a key tool for important links such as gene analysis and disease diagnosis, and its performance directly affects the reliability of the detection results. With the development of precision medical technology, higher requirements are put forward for the spotting accuracy and hybridization reaction effect of the flow-through hybridization analysis platform, and how to realize uniform distribution and accurate quantification of probes has become a research focus in this field.

[0003] At present, the traditional flow-through hybridization analysis platform mainly adopts two methods of contact spotting and non-contact spotting. The contact spotting directly applies the probe solution to the surface of the chip, but this method causes the probe to concentrate and overlap in the center of the spotting area, gradually sparse outward and present uneven distribution. The non-contact spotting improves the probe distribution problem by combining droplets into a circle, but due to the lack of a reasonable droplet distribution algorithm, the droplet arrangement is often too compact, and the gap is too small, which is prone to adhesion. The above two spotting methods, the former affects the accuracy and readability of the hybridization reaction results, and the latter causes uneven hybridization color development, neither of which can realize accurate quantification and digital detection, and both have the defects of low detection accuracy and low result readability. SUMMARY

[0004] To overcome the defects of low detection accuracy and low result readability of the existing flow-through hybridization analysis platform, the present application proposes the following technical solutions:

[0005] In a first aspect, the present application proposes a flow-through hybridization analysis method based on, comprising:

[0006] Inkjet spotting is performed on the paper-based chip to generate a circular spot array; wherein each circular spot is composed of a plurality of droplets;

[0007] The spatial filling coefficient of the droplets is calculated, and if the spatial filling coefficient is less than a set threshold, the spatial filling coefficient is optimized until the spatial filling coefficient is not less than the threshold, and a paper-based chip with uniform droplet distribution is obtained;

[0008] The paper-based chip with uniform droplet distribution is subjected to flow-through hybridization reaction to obtain a paper-based chip carrying hybridization signals;

[0009] The paper-based chip carrying hybridization signals is subjected to qualitative and quantitative detection analysis to obtain an analysis result.

[0010] As a preferred technical solution, inkjet spotting is performed on the paper-based chip to generate a circular spot array, comprising:

[0011] determining the number of microdroplets to be generated, generating a floating point array of corresponding length;

[0012] adding an offset value to each element of the floating point array to obtain a pre-processed floating point array;

[0013] based on the golden angle rule, calculating the polar angle of each microdroplet using the pre-processed floating point array, and adjusting the polar radius of each microdroplet through a square root function to obtain the polar coordinates of each microdroplet;

[0014] converting the polar coordinates of each microdroplet into Cartesian coordinates to obtain the distribution coordinates of the microdroplets on the paper-based chip;

[0015] The inkjet spotter spots the paper-based chip according to the distribution coordinates to form an array of circular spots.

[0016] As a preferred technical solution, the spatial filling coefficient of the microdroplets is calculated, including:

[0017] According to the distribution coordinates of the microdroplets on the paper-based chip, a Voronoi diagram is constructed;

[0018] Obtaining the Voronoi partition information corresponding to each microdroplet, and screening the Voronoi partitions in the limited area;

[0019] Calculating the standard deviation of the area of each limited area Voronoi partition;

[0020] According to the total area of the circular spots and the number of microdroplets, the ideal average area is calculated;

[0021] Based on the ratio of the ideal average area to the statistical standard deviation, the spatial filling coefficient of the microdroplets is determined.

[0022] As a preferred technical solution, the spatial filling coefficient is optimized until the spatial filling coefficient is not less than a threshold value, to obtain a paper-based chip with uniform microdroplet distribution, including:

[0023] Calculating the centroid coordinates of the Voronoi partitions in the limited area as new microdroplet distribution coordinates;

[0024] According to the new microdroplet distribution coordinates, iteratively calculating the spatial filling coefficient of the microdroplets until the spatial filling coefficient is not less than a set threshold value, and terminating the iteration to obtain a paper-based chip with uniform microdroplet distribution.

[0025] As a preferred technical solution, the paper-based chip carrying hybridization signals is subjected to qualitative detection analysis, including:

[0026] Scanning the paper-based chip carrying hybridization signals to obtain an image containing circular spots;

[0027] The trained machine learning model is used to identify the positive and negative of the circular spot in the image.

[0028] As a preferred technical solution, the paper-based chip carrying hybrid signals is subjected to quantitative detection analysis, comprising:

[0029] The microdroplets in the circular spot in the image are identified, and the number and gray intensity of the microdroplets are counted;

[0030] Based on the number and gray intensity of the microdroplets, combined with the microdroplet distribution characteristics or standard reference data, the quantitative detection value of the target biomolecule is calculated and output.

[0031] As a preferred technical solution, before identifying the microdroplets in the circular spot in the image and counting the number and gray intensity of the microdroplets, the method further comprises correcting the direction of the image, comprising:

[0032] The center partition polygon of the circular spot in the image is extracted, and the centroid of the center partition polygon is calculated;

[0033] The vertex coordinates of the center partition polygon are converted into coordinates relative to the centroid;

[0034] Based on the coordinates relative to the centroid, the moment of inertia of the center partition polygon is calculated;

[0035] According to the moment of inertia, the actual principal axis direction angle of the center partition is calculated;

[0036] According to the actual principal axis direction angle, the image is rotated and corrected.

[0037] In a second aspect, the present application also provides a hybrid analysis device based on flow guiding, which is applied to the hybrid analysis method based on flow guiding according to any one of the first aspect, comprising:

[0038] The spotting module is used for inkjet spotting on the paper-based chip to generate an array of circular spots; wherein each circular spot is composed of a plurality of microdroplets;

[0039] The calculation module is used for calculating the spatial filling coefficient of the microdroplets, and if the spatial filling coefficient is less than a set threshold, the spatial filling coefficient is optimized until the spatial filling coefficient is not less than the threshold, so as to obtain the paper-based chip with uniform microdroplet distribution;

[0040] The hybrid module is used for carrying out flow hybrid reaction on the paper-based chip with uniform microdroplet distribution to obtain the paper-based chip carrying hybrid signals;

[0041] The analysis module is used for carrying out qualitative and quantitative detection analysis on the paper-based chip carrying hybrid signals to obtain the analysis result.

[0042] In a third aspect, the present application also provides an electronic device, comprising a memory, a processor, and a computer program stored in the memory and executable on the processor, wherein the processor implements the operations performed by the method for hybridization analysis by flow guiding according to any one of the first aspect.

[0043] In a fourth aspect, the present application also provides a computer readable storage medium, which stores a program, and the program is executed by a processor to perform the operations performed by the method for hybridization analysis by flow guiding according to any one of the first aspect.

[0044] The present application has at least the following beneficial effects:

[0045] The present application forms a circular spot array composed of microdroplets by inkjet spotting on a paper-based chip, and realizes uniform distribution of microdroplets in the circular spot by calculating and optimizing the space filling coefficient; the uniform distribution constructs a consistent reaction environment for the hybridization reaction, guarantees the stability and repeatability of the hybridization signal, carries out qualitative and quantitative detection analysis based on the stable hybridization signal, reduces the signal difference interference between microdroplets, thereby improving the detection accuracy of the hybridization analysis by flow guiding, and at the same time makes the signal distribution of the circular spot more regular, and enhances the readability of the results. BRIEF DESCRIPTION OF DRAWINGS

[0046] Figure 1 The flowchart of the method for hybridization analysis by flow guiding provided by the embodiments of the present application.

[0047] Figure 2 (a) is the distribution diagram of 5000 microdroplets generated by the Fibonacci spiral method provided by the embodiments of the present application.

[0048] Figure 2 (b) is the distribution diagram of 10000 microdroplets generated by the Fibonacci spiral method provided by the embodiments of the present application.

[0049] Figure 3 (a) is the macroscopic schematic diagram of the initial distribution of microdroplets provided by the embodiments of the present application.

[0050] Figure 3 (b) is the macroscopic schematic diagram of the distribution of microdroplets after optimization by the Lloyd algorithm provided by the embodiments of the present application.

[0051] Figure 4 (a) is the microscopic schematic diagram of the initial distribution of microdroplets provided by the embodiments of the present application.

[0052] Figure 4 (b) is the microscopic schematic diagram of the distribution of microdroplets after optimization by the Lloyd algorithm provided by the embodiments of the present application.

[0053] Figure 5 A schematic diagram of an architecture of a hybridization analysis device based on flow guiding provided by an embodiment of the present application.

[0054] Figure 6 A structural schematic diagram of an electronic device provided by an embodiment of the present application. DETAILED DESCRIPTION

[0055] Other advantages and effects of the present application can be easily understood by those skilled in the art from the content disclosed in the present specification. The present application can also be implemented or applied by means of other different specific embodiments, and each detail in the present specification can be modified or changed based on different views and applications without departing from the spirit of the present application. It should be understood that the preferred embodiments are only for illustrating the present application, but not for limiting the protection scope of the present application.

[0056] It should be noted that the diagrams provided in the following embodiments only schematically illustrate the basic concept of the present application, and thus the diagrams only show the components related to the present application, but are not drawn according to the number, shape and size of the components in actual implementation, and the type, number and ratio of each component in actual implementation can be arbitrarily changed, and the layout type of the components can also be more complex.

[0057] In the following description, a large number of details are discussed to provide a more thorough explanation of the embodiments of the present application, however, it is obvious for those skilled in the art that the embodiments of the present application can be implemented without these specific details, and in other embodiments, the well-known structures and devices are shown in the form of block diagrams instead of details, to avoid making the embodiments of the present application difficult to understand.

[0058] Embodiment 1

[0059] The present embodiment proposes a flow guiding hybridization analysis method, as shown in Figure 1 Figure 1 A flowchart of a flow guiding hybridization analysis method provided by the present embodiment, the method comprises the following steps:

[0060] S1: Inkjet spotting is performed on a paper-based chip to generate an array of circular spots; wherein each circular spot is composed of a plurality of droplets;

[0061] S2: The spatial filling coefficient of the droplets is calculated, if the spatial filling coefficient is less than a set threshold, the spatial filling coefficient is optimized until the spatial filling coefficient is not less than the threshold, to obtain a paper-based chip with uniform droplet distribution;

[0062] S3: The paper-based chip with uniform droplet distribution is subjected to flow guiding hybridization reaction to obtain a paper-based chip carrying hybridization signals; ​

[0063] S4: Qualitative and quantitative detection analysis is performed on the paper-based chip carrying the hybridization signal to obtain an analysis result.

[0064] In the specific implementation process, first, a circular spot array is generated on the paper-based chip by an inkjet spotting instrument according to a preset number of droplets in combination with a golden angle spiral algorithm, so that the droplets are preliminarily arranged; then, a spatial filling coefficient of the droplets is calculated, if the coefficient is less than a set threshold, Lloyd algorithm is used for iterative optimization, a Voronoi diagram of the droplet distribution is constructed, the center of each partition is calculated and used as a new droplet coordinate, the Voronoi diagram is repeatedly constructed and the coordinate is repeatedly updated, until the spatial filling coefficient is not less than the threshold, so as to ensure that the droplets are uniformly dispersed and the spacing is reasonable in the circular spot; subsequently, the optimized paper-based chip is placed in a flow-through hybridization reaction device, a reaction solution containing target biomolecules is introduced, and the reaction solution uniformly flows through the droplets by the flow-through structure of the chip, so as to complete the hybridization reaction of the probe and the target; finally, the paper-based chip carrying the hybridization signal is scanned and imaged, the center partition polygon of the circular spot is extracted, the center of gravity is calculated, the inertia moment is derived to determine the main axis direction and correct the image, then the machine learning model is used to identify the positive and negative of the droplets, the number and gray intensity of the droplets are counted, the quantitative analysis is realized by combining the droplet distribution characteristics or standard product data, and the accuracy and readability of the flow-through hybridization detection result are improved through the standardization of the distribution optimization and the image correction.

[0065] Embodiment 2

[0066] This embodiment is improved on the basis of the flow-through hybridization analysis method proposed in Embodiment 1.

[0067] Optionally, the paper-based chip is subjected to inkjet spotting to generate a circular spot array, comprising:

[0068] The number of droplets to be generated is determined, and a floating point array with a corresponding length is generated;

[0069] An offset value is added to each element of the floating point array to obtain a preprocessed floating point array;

[0070] Based on the golden angle rule, the polar angle of each droplet is calculated by using the preprocessed floating point array, and the polar radius of each droplet is adjusted by a square root function to obtain the polar coordinates of each droplet;

[0071] The polar coordinates of each droplet are converted into Cartesian coordinates to obtain the distribution coordinates of the droplets on the paper-based chip;

[0072] The paper-based chip is spotted by the inkjet spotting instrument according to the distribution coordinates to form a circular spot array.

[0073] In an example, as Figure 2 (a) and Figure 2(b) as shown, the microdroplet distribution is generated by Fibonacci spiral method: determine the number of microdroplets to be generated N (such as N > 2000), generate an initial floating point array, superimpose an offset value of 0.5 on each element in the array to obtain a preprocessed floating point array Indices = [0, 1, 2,..., N-1] + 0.5, and eliminate the central point symmetry overlap.

[0074] Set the maximum radius of the circular spot as R, and calculate the polar angle of each microdroplet based on the golden angle rule Wherein is the golden angle radian proportion factor.

[0075] Calculate the polar radius p of each microdroplet by the square root function to compensate for the nonlinear expansion of the polar coordinate area:

[0076]

[0077] Convert the polar coordinates to Cartesian coordinates:

[0078] X i =r i ·cos(θ i )

[0079] y i =r i ·sin(θ i )

[0080] Control the inkjet spotter to have a resolution ≥1200 dpi, a precision of 0.1-1.0 nL of spotting volume, and spot on the paper-based chip according to the Cartesian coordinates to form a circular spot array with a total area of 3 mm 2 , so that the microdroplets are arranged according to the rule of no intersection.

[0081] In this embodiment, the microdroplet distribution is generated by the Fibonacci spiral method, and the golden angle distribution and polar radius adjustment are used to ensure that the microdroplets are uniformly distributed inside the large circle. This method not only avoids the problem of uneven distribution of probes, but also ensures the appropriate spacing between each microdroplet, thereby effectively preventing the occurrence of adhesion phenomenon.

[0082] Optionally, the spatial filling coefficient of the microdroplets is calculated, including:

[0083] According to the distribution coordinates of the microdroplets on the paper-based chip, a Voronoi diagram is constructed;

[0084] Obtain the Voronoi partition information corresponding to each microdroplet, and screen out the Voronoi partitions in the limited area;

[0085] Calculate the standard deviation of the area of each limited area Voronoi partition;

[0086] Calculate the ideal average area based on the total area of ​​the circular spots and the number of droplets;

[0087] The space-filling coefficient of a droplet is determined based on the ratio of the ideal average area to the statistical standard deviation.

[0088] In one example, a Voronoi diagram is constructed based on the droplet distribution coordinates, and the partition information corresponding to each droplet is extracted. A finite number of regions is selected from the partition information that do not contain -1 and have a valid number of vertices {len(region)}>0. For each finite region, the vertex coordinates are obtained, and then processed using the formula... Calculate the area of ​​the partition, where (xi,yi) represents the vertex coordinates of the polygon.

[0089] Then, the standard deviation of the area of ​​the finite region is calculated; the ideal average area is obtained by dividing the total area of ​​the circular spots by the number of droplets; the ideal average area is compared with the standard deviation to determine the space filling factor.

[0090] Optionally, the space fill factor is optimized until it is not less than a threshold, resulting in a paper-based chip with uniform droplet distribution, including:

[0091] The centroid coordinates of the Voronoi partitions of the finite region are calculated as the new droplet distribution coordinates;

[0092] The space-filling coefficient of the microdroplets is calculated iteratively based on the new microdroplet distribution coordinates. The iteration is terminated when the space-filling coefficient is not less than a set threshold, resulting in a paper-based chip with uniform microdroplet distribution.

[0093] In one example, the number of iterations is set (e.g., 10 times), with the initial distribution coordinates of the droplets as input; during the cyclic iteration, a Voronoi diagram is constructed, and valid vertices are selected ( A finite region (where len(region)>0) is defined. For each finite region, the centroid of the vertex coordinates is calculated. If a clipping radius is set, when the distance of the centroid from the center exceeds the radius, the centroid is scaled proportionally to the boundary. All centroids are used as the coordinates of the new droplet distribution, and the Voronoi diagram and centroid update operation are repeated. After each iteration, the space filling coefficient is calculated until the coefficient is not less than the set threshold, at which point the iteration is terminated.

[0094] In practical calculations, the density of the fill is quantified by comparing the difference between the area of ​​each Voronoi cell and the ideal average area. The space fill factor can be calculated using the following formula:

[0095]

[0096] In the formula, IdealAverageArea represents the ideal average area of each point, which is equal to the total area divided by the number of points. StandardDeviationofVoronoiAreas represents the standard deviation of the area of the Voronoi cell of each point. The larger the space filling coefficient, the more uniform the distribution of the point set, and the better the filling effect. If the standard deviation is large, some areas are too dense or sparse, resulting in poor filling effect.

[0097] In this embodiment, as shown in Figure 3 (a) and Figure 3 (b) shown, the space filling coefficient is introduced to evaluate the uniformity of the droplet distribution, and the Lloyd algorithm is used to optimize the space filling coefficient of the distribution that does not meet the set threshold. This step can significantly improve the uniformity of the droplet distribution, so that each droplet can get sufficient hybridization reaction conditions, thereby improving the accuracy and repeatability of the detection results. Figure 4 Figure 4 Optionally, the paper-based chip carrying the hybridization signal is subjected to qualitative detection analysis, including:

[0098] The paper-based chip carrying the hybridization signal is scanned to obtain an image containing circular spots;

[0099] The trained machine learning model is used to identify the positive and negative of the circular spots in the image.

[0100] In an example, the paper-based chip is scanned to obtain a large image containing circular spots and generate a thumbnail, the center partition features of the circular spots in the large image are extracted, a training data set containing one hundred thousand labeled positive spots is constructed, a machine learning model is trained, the thumbnail is preprocessed, the trained model is used to identify the positive and negative of the circular spots, the position and probability of the positive spots are output, and the transparency and gray intensity of the positive spots in the thumbnail are calculated by computer vision.

[0101] Optionally, the paper-based chip carrying the hybridization signal is subjected to quantitative detection analysis, including:

[0102] The number and gray intensity of the droplets in the circular spots in the image are identified and counted;

[0103] Based on the number and gray intensity of the droplets, combined with the droplet distribution characteristics or standard reference data, the quantitative detection value of the target biomolecule is calculated and output.

[0104] Optionally, before identifying the droplets in the circular spots in the image and counting the number and gray intensity of the droplets, the method further includes correcting the direction of the image, including:

[0105] Optionally, before identifying the droplets in the circular spots in the image and counting the number and gray intensity of the droplets, the method further includes correcting the direction of the image, including:

[0106] ​extracting a center-partition polygon of the circular spot in the image, and calculating a centroid of the center-partition polygon;

[0107] converting vertex coordinates of the center-partition polygon into coordinates relative to the centroid;

[0108] calculating moments of inertia of the center-partition polygon based on the coordinates relative to the centroid;

[0109] calculating an actual principal axis direction angle of the center-partition based on the moments of inertia;

[0110] performing a rotation correction on the image according to the actual principal axis direction angle.

[0111] In an example, a center-partition polygon of a circular spot in an image is extracted, and a centroid (C x , C y ) is calculated. Vertex coordinates of the center-partition polygon are converted into coordinates relative to the centroid x i ′ =x i -C x and y i ′ =y i -C y ; based on the relative coordinates, moments of inertia I xx along the x-axis, I yy along the y-axis, and I xy of the center-partition polygon are calculated:

[0112]

[0113] An actual principal axis direction angle of the center-partition is calculated through the moments of inertia:

[0114]

[0115] The center-partition polygon corresponding to the microdroplet distribution is retrieved, and the centroid, the relative coordinates, and the moments of inertia are repeatedly calculated to obtain a design principal axis direction angle θ0.

[0116] A deviation Δθ=θ-θ0 between the actual direction angle and the design direction angle is calculated, and the image is rotated according to the deviation.

[0117] In the example, through recording of partition information of each point, especially the center-partition features, the direction of the chip can be quickly identified, and the chip can be rotated as necessary. This process simplifies subsequent image processing steps, reduces unnecessary complexity and computational load, and greatly improves the speed and efficiency of data analysis.

[0118] Example 3

[0119] As Figure 5As shown, the embodiment proposes a device for flow-through hybridization analysis, which is applied to the method for flow-through hybridization analysis as described in the above embodiment, and includes a spotting module 100, a calculation module 200, a hybridization module 300 and an analysis module 400.

[0120] The spotting module 100 is configured to perform inkjet spotting on the paper-based chip to generate a circular spot array; each circular spot is composed of a plurality of droplets; the calculation module 200 is configured to calculate the spatial filling coefficient of the droplets, and if the spatial filling coefficient is less than a set threshold, the spatial filling coefficient is optimized until the spatial filling coefficient is not less than the threshold, thereby obtaining a paper-based chip with uniform droplet distribution; the hybridization module 300 is configured to perform flow-through hybridization reaction on the paper-based chip with uniform droplet distribution, thereby obtaining a paper-based chip carrying hybridization signals; and the analysis module 400 is configured to perform qualitative and quantitative detection analysis on the paper-based chip carrying hybridization signals, thereby obtaining an analysis result.

[0121] It should be noted that the above explanation of the method for flow-through hybridization analysis is also applicable to the device for flow-through hybridization analysis, and thus will not be repeated here.

[0122] Embodiment 4

[0123] Figure 6 The electronic device 500 provided in the embodiment is shown in a structural schematic diagram. The electronic device 500 includes a memory 501, a processor 502, and a computer program stored in the memory 501 and executable on the processor 502.

[0124] The processor 502 executes the program to implement the method for flow-through hybridization analysis provided in the above embodiment.

[0125] Further, the electronic device 500 further includes a communication interface 503 for communication between the memory 501 and the processor 502.

[0126] The memory 501 can include a high-speed RAM (Random Access Memory) memory, and can also include a non-volatile memory, such as at least one disk memory.

[0127] If the memory 501, the processor 502 and the communication interface 503 are implemented independently, the communication interface 503, the memory 501 and the processor 502 can be connected with each other through a bus and complete communication between each other. The bus can be an ISA (Industry Standard Architecture) bus, a PCI (Peripheral Component) bus or an EISA (Extended Industry Standard Architecture) bus, etc. The bus can be divided into an address bus, a data bus, a control bus, etc. For the convenience of representation, Figure 6 Only one thick line is used in the figure to represent the bus, but it does not mean that there is only one bus or only one type of bus.

[0128] Optionally, in a specific implementation, if the memory 501, the processor 502 and the communication interface 503 are integrated on a chip, the memory 501, the processor 502 and the communication interface 503 can complete communication between each other through an internal interface.

[0129] The processor 502 can be a CPU (Central Processing Unit) or an ASIC (Application Specific Integrated Circuit) or one or more integrated circuits configured to implement embodiments of the present application.

[0130] The embodiment of the present application also provides a computer readable storage medium, which stores a computer program, and the program is executed by a processor to implement the above-mentioned method for analyzing based on flow-through hybridization.

[0131] In the description of the present specification, the description of the terms "one embodiment", "some embodiments", "an example", "a specific example" or "some examples" means that the specific features, structures, materials or characteristics described in connection with the embodiment or example are included in at least one embodiment or example of the present application. In the present specification, the illustrative description of the above terms is not necessarily for the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described can be combined in any one or N embodiments or examples in a suitable manner. In addition, the skilled in the art can combine and combine the different embodiments or examples described in the present specification and the features of the different embodiments or examples without contradiction.

[0132] Furthermore, the terms "first", "second", or the like, are used only to describe the purpose of distinguishing between different features, and are not intended to imply or suggest relative importance of, or a hierarchy between, the indicated features. Thus, a feature defined with "first" or "second" can implicitly or explicitly include at least one of the other feature. The meaning of "N" is at least two, such as two, three, etc., unless explicitly specified otherwise.

[0133] Any process or method described in a flowchart or otherwise described herein can be understood as representing code modules, segments, or portions of code that include one or more executable instructions for implementing specific logic functions (or steps) of the process, and the various embodiments of the present application can include additional or fewer steps or processes in comparison to those shown in the figures. It should also be understood that the sequences of processes depicted in the figures, and described herein, can not be restricted to the order in which the processes are illustrated, and that many variations can be possible.

[0134] It should be understood that various parts of the present application can be implemented in hardware, software, firmware or a combination thereof. In the above embodiments, the various steps or methods can be implemented in software or firmware that is stored in memory and executed by a suitable instruction execution system. As such, in some embodiments, the memory can include computer readable media in the form of a storage medium or memory media for holding data and / or instructions accessible by a processor or computing platform. Non-limiting examples of computer readable media include random access memory (RAM), read only memory (ROM), programmable read only memory (PROM), erasable programmable read only memory (EPROM), electrically erasable programmable read only memory (EEPROM), flash memory, compact disc read only memory (CD-ROM), digital versatile discs (DVDs), magnetic disks, and tapes.

[0135] It should be understood that any number of processes can be implemented using the processes described above, and that modifications can be made by persons of ordinary skill in the art to adapt the processes for use with other processes or systems without departing from the scope of the present application. Those of ordinary skill in the art can implement the processes described above, and / or portions thereof, in software programs or computer programs. The software programs or computer programs can be stored in any appropriate non-transitory computer-readable data storage media including a storage media that is external to a computer or computing platform or that is internal to a computer or computing platform. Examples of internal computer-readable storage media include common RAM, ROM, flash memory, etc. Examples of external computer-readable storage media include optical, magnetic, and electronic storage media, including, but not limited to, disk drives, magnetic disks, magnetic tape, CD-ROMs, DVDs, and semiconductor memory devices.

[0136] Obviously, the above-described embodiments of the present application are only examples for clearly illustrating the present application, and are not intended to limit the embodiments of the present application. Based on the above description, other different forms of changes or variations can be made by those of ordinary skill in the art. Here, it is not necessary or possible to exhaust all the embodiments. Any modifications, equivalent replacements, and improvements, etc. made within the spirit and principle of the present application shall be included in the protection scope of the claims of the present application.

Claims

1. A method of fluid-directed hybridization analysis, characterized by, The method comprises the following steps: inkjet spotting on a paper-based chip to generate a circular spot array, wherein each circular spot is composed of a plurality of droplets, including: determining the number of droplets to be generated, generating a floating point array of corresponding length; adding an offset value to each element of the floating point array to obtain a preprocessed floating point array; based on the golden angle rule, using the preprocessed floating point array to calculate the polar angle of each droplet, and adjusting the polar radius of each droplet through a square root function to obtain the polar coordinates of each droplet; convert the polar coordinates of each droplet into Cartesian coordinates to obtain the distribution coordinates of the droplets on the paper-based chip; the inkjet spotter spots the paper-based chip according to the distribution coordinates to form a circular spot array; construct a Voronoi diagram according to the distribution coordinates of the droplets on the paper-based chip; obtain the Voronoi partition information corresponding to each droplet, and select the Voronoi partition in the limited area; calculate the standard deviation of the area of each limited area Voronoi partition; calculate the ideal average area according to the total area of the circular spot and the number of droplets; determine the space filling coefficient of the droplet based on the ratio of the ideal average area to the statistical standard deviation; if the space filling coefficient is less than the set threshold, optimize the space filling coefficient until the space filling coefficient is not less than the threshold, and obtain a paper-based chip with uniform droplet distribution; perform a flow-through hybridization reaction on the paper-based chip with uniform droplet distribution to obtain a paper-based chip carrying hybridization signals; perform qualitative and quantitative detection analysis on the paper-based chip carrying hybridization signals to obtain analysis results.

2. The flow-through hybridization assay method according to claim 1, wherein Optimize the space filling coefficient until the space filling coefficient is not less than the threshold to obtain a paper-based chip with uniform droplet distribution, including: calculate the centroid coordinates of the Voronoi partition in the limited area as the new droplet distribution coordinates; iteratively calculate the space filling coefficient of the droplet according to the new droplet distribution coordinates until the space filling coefficient is not less than the set threshold, and terminate the iteration to obtain a paper-based chip with uniform droplet distribution.

3. The flow-through hybridization assay method according to claim 1, wherein The qualitative detection analysis of the paper-based chip carrying hybridization signals includes: scanning the paper-based chip carrying hybridization signals to obtain an image containing circular spots; using a trained machine learning model to identify the positive and negative of the circular spots in the image.

4. The flow-through hybridization assay method according to claim 3, wherein The quantitative detection analysis of the paper-based chip carrying hybridization signals includes: identify the droplets in the circular spots in the image, and count the number and gray intensity of the droplets; based on the number and gray intensity of the droplets, combined with the droplet distribution characteristics or standard reference data, calculate and output the quantitative detection value of the target biomolecule.

5. The flow-through hybridization assay method according to claim 4, wherein Before identifying the droplets in the circular spots in the image, counting the number and gray intensity of the droplets, the method further includes correcting the direction of the image, including: extract the center partition polygon of the circular spot in the image, and calculate the centroid of the center partition polygon; convert the vertex coordinates of the center partition polygon to coordinates relative to the centroid; based on the coordinates relative to the centroid, calculate the moment of inertia of the center partition polygon; calculate the actual principal axis direction angle of the center partition according to the moment of inertia; rotate the image according to the actual principal axis direction angle.

6. A flow-through hybridization assay device for use in the flow-through hybridization assay method according to any one of claims 1 to 5, characterized by ​ A spotting module is configured to perform inkjet spotting on the paper-based chip to generate a circular spot array, wherein each circular spot is composed of a plurality of droplets; A computing module is configured to calculate a spatial filling coefficient of the droplets, and if the spatial filling coefficient is less than a set threshold, the spatial filling coefficient is optimized until the spatial filling coefficient is not less than the threshold, thereby obtaining a paper-based chip with uniform droplet distribution; A hybridization module is configured to perform a flow-through hybridization reaction on the paper-based chip with uniform droplet distribution, thereby obtaining a paper-based chip carrying a hybridization signal; An analysis module is configured to perform qualitative and quantitative detection analysis on the paper-based chip carrying the hybridization signal, thereby obtaining an analysis result.

7. An electronic device, comprising: The electronic device includes a memory, a processor, and a computer program stored on the memory and running on the processor, wherein the processor executes the program to implement the operations performed by the flow-through hybridization analysis method according to any one of claims 1 to 5.

8. A computer-readable storage medium, characterized in that, The computer readable storage medium stores a program, and the program is executed by the processor to perform the operations performed by the flow-through hybridization analysis method according to any one of claims 1 to 5.

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