A method for determining the luminescence intensity of an electrochemiluminescence array sensor image

Abstract

The application discloses a method and device for determining the light-emitting intensity of an electrochemiluminescence array sensor image, and relates to the technical field of image processing. A terminal imaging image of an electrochemiluminescence array sensor is acquired, and a background image in the terminal imaging image is removed to obtain a sample image; the sample image is rotated by a preset rotation angle with the center of the sample image as a rotation point, so that the light-emitting area of the sample image is parallel or perpendicular along the horizontal direction; according to the rotated sample image, coordinate data of the light-emitting area is extracted; according to the coordinate data of the light-emitting area, the gray value of the light-emitting area is determined, and the total gray value of all light-emitting areas is determined as the light-emitting intensity of the terminal imaging image. The method can quickly and accurately count the light-emitting intensity of the electrochemiluminescence array sensor image.

Description

Technical Field

[0001] This invention relates to the field of image processing technology, and in particular to a method and apparatus for determining the luminescence intensity of an image from an electrochemiluminescence array sensor. Background Technology

[0002] Currently, disease biomarkers (such as specific proteins, nucleic acids, metabolites, and cytokines) serve as important clinical indicators for disease diagnosis, treatment, and prognosis. High-throughput, high-sensitivity, and rapid detection is a core task in clinical diagnosis and monitoring. Therefore, early, accurate, rapid, and affordable disease diagnosis is crucial for improving patient treatment and prognosis, as well as enhancing public health and hygiene management.

[0003] In recent years, luminescent array biosensors have become a cutting-edge solution for addressing the challenges of high-sensitivity multi-channel detection. The array design allows for the simultaneous detection of multiple biomarkers on a single test strip / chip, significantly improving throughput and detection efficiency. The luminescent signals (chemiluminescent spots, fluorescent dot arrays, or ECL spatial distribution) generated by the array biosensor need to be captured as digital images by an optical imaging system (typically using low-cost, miniaturized CMOS or mobile phone image sensors). Accurate quantification of the luminescence intensity is then fundamental to obtaining biomarker concentrations. Currently, the extraction of light intensity from images from luminescent immunoassay array sensors mainly relies on image processing software like ImageJ, involving a series of complex settings and operations for statistical analysis. This method is time-consuming and generally inaccurate, sometimes resulting in missed entries or incorrect calculations. Accurate extraction of the luminescent region from an imaging image typically takes 2-10 minutes and requires specialized training, severely limiting the research and practical application of luminescent array sensors.

[0004] Therefore, there is an urgent need for a method that can quickly and accurately count the luminescence intensity of images from electrochemiluminescence array sensors. Summary of the Invention

[0005] Therefore, it is necessary to provide a method and apparatus for determining the luminescence intensity of an electrochemiluminescence array sensor image to address the above-mentioned technical problems. This method can quickly and accurately count the luminescence intensity of an electrochemiluminescence array sensor image.

[0006] The present invention adopts the following technical solution:

[0007] This invention provides a method for determining the luminescence intensity of an image from an electrochemiluminescence array sensor, comprising:

[0008] Acquire the terminal imaging image of the electrochemiluminescence array sensor, and remove the background image from the terminal imaging image to obtain the sample image;

[0009] Using the center of the sample image as the rotation point, rotate the sample image by a preset rotation angle so that the luminous area of ​​the sample image is parallel or perpendicular to the horizontal direction;

[0010] Based on the rotated sample image, extract the coordinate data of the luminescent region;

[0011] Based on the coordinate data of the luminous area, the gray value of the luminous area is determined, and the total gray value of all luminous areas is determined as the luminous intensity of the terminal imaging image.

[0012] Optionally, the sample image is rotated by a preset rotation angle with the center of the sample image as the rotation point, including:

[0013] Determine the center of the rotated sample image based on the center of the sample image and the rotation angle;

[0014] Based on the center of the sample image and the center of the rotated sample image, the new pixels generated by the rotation are assigned a value of 0 to obtain the rotated sample image.

[0015] Optionally, based on the rotated sample image, the coordinate data of the luminescent region is extracted, including:

[0016] Based on the rotated sample image, the luminous area of ​​the rotated sample image is determined through the human-computer interaction interface;

[0017] Determine if the selected luminous area is correct. If the luminous area is selected correctly, extract the coordinate data of the luminous area.

[0018] Optionally, determining whether the light-emitting area has been selected correctly includes:

[0019] Obtain the grayscale values ​​of all pixels on the boundary line of the luminous region, the average grayscale value of the non-luminous region, and three times the standard deviation of the non-luminous region;

[0020] If the gray values ​​of all pixels on the boundary line of the luminous region are less than the sum of the average gray value of the non-luminous region and three times the standard deviation, then the luminous region is determined to be correctly selected.

[0021] Otherwise, the luminous area has been selected incorrectly.

[0022] Optionally, in the event of an incorrect selection of the luminescent region, the method further includes:

[0023] Reselect the luminous region of the rotated sample image until the luminous region is correctly selected.

[0024] Optionally, the rotation angle is determined in advance by adjusting historical terminal imaging images of the electrochemiluminescence array sensor.

[0025] Optionally, the direction of rotation of the sample image is determined by the sign of the rotation angle.

[0026] This invention provides a device for determining the luminescence intensity of an image from an electrochemiluminescence array sensor, comprising:

[0027] The acquisition module is used to acquire the terminal imaging image of the electrochemiluminescence array sensor and remove the background image from the terminal imaging image to obtain the sample image;

[0028] The rotation module is used to rotate the sample image by a preset rotation angle with the center of the sample image as the rotation point, so that the luminous area of ​​the sample image is parallel or perpendicular to the horizontal direction.

[0029] The extraction module is used to extract the coordinate data of the luminescent region based on the rotated sample image;

[0030] The determination module is used to determine the grayscale value of the luminous area based on the coordinate data of the luminous area, and to determine the total grayscale value of all luminous areas as the luminous intensity of the terminal imaging image.

[0031] The present invention provides a computer-readable storage medium storing a computer program that, when executed by a processor, implements the above-described method for determining the luminescence intensity of an electrochemiluminescence array sensor image.

[0032] The present invention provides a computer device, including a memory, a processor, and a computer program stored in the memory and executable on the processor. When the processor executes the program, it implements the above-described method for determining the luminescence intensity of an electrochemiluminescence array sensor image.

[0033] The above-mentioned at least one technical solution adopted in this invention can achieve the following beneficial effects:

[0034] In this invention, the background of the terminal imaging image is first removed to reduce the interference of background noise on the detection of the luminous region. Furthermore, since the original luminous region is tilted, subsequent selection of the luminous region is easily affected by interference from other regions, making accurate selection of the luminous region's position very difficult. Therefore, this invention rotates the sample image obtained after removing the background image to ensure that the luminous region of the image is parallel to the horizontal or vertical direction, improving the accuracy of luminous region selection and thus resulting in higher accuracy of the luminous intensity of the terminal imaging image. Moreover, this invention uses background separation, rotation, and coordinate extraction to determine the luminous intensity of the image, simplifying the calculation process, reducing redundant operations, and improving the calculation speed of the image's luminous intensity. Attached Figure Description

[0035] The accompanying drawings, which are included to provide a further understanding of the invention and form part of this invention, illustrate exemplary embodiments of the invention and are used to explain the invention, but do not constitute an undue limitation of the invention. In the drawings:

[0036] Figure 1 A schematic flowchart of a method for determining the luminescence intensity of an image from an electrochemiluminescence array sensor provided by the present invention;

[0037] Figure 2 This invention provides an image processing schematic diagram;

[0038] Figure 3 A visual schematic diagram of a Matlab command window provided by the present invention;

[0039] Figure 4 A flowchart illustrating the implementation of a method for determining the luminescence intensity of an image from an electrochemiluminescence array sensor, provided by this invention.

[0040] Figure 5 This is a schematic diagram of a computer device for determining the luminescence intensity of an image from an electrochemiluminescence array sensor, as provided by the present invention. Detailed Implementation

[0041] To make the objectives, technical solutions, and advantages of this invention clearer, the technical solutions of this invention will be clearly and completely described below in conjunction with specific embodiments and corresponding drawings. Obviously, the described embodiments are only a part of the embodiments of this invention, and not all of them. Based on the embodiments of this invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this invention.

[0042] Early, accurate, rapid, and affordable disease diagnosis is crucial for improving patient treatment and prognosis, as well as public health and hygiene management. However, achieving this goal faces significant challenges: ①. The need for extremely low concentration detection: Many key biomarkers (such as early cancer markers, cardiovascular risk factors, and infectious disease pathogen antigens / antibodies) are present in bodily fluids (blood, saliva, urine, etc.) in the early stages of disease, but their concentrations are often at ultra-trace levels, ranging from picomolars (pM) to femtomolars (fM) or even lower. ②. The need for high-throughput detection: The diagnostic value of a single biomarker is usually limited; combining multiple biomarkers (multiplexing) can significantly improve diagnostic accuracy (specificity and sensitivity) and disease classification capabilities (such as differentiating viral subtypes and cancer stages). ③. The need for speed and convenience: In point-of-care testing (POCT) and resource-constrained scenarios (such as home surveillance and epidemiological field investigations), there is an urgent need for miniaturized, automated, and user-friendly devices to achieve "sample in - result out".

[0043] Traditional central laboratory tests (such as enzyme-linked immunosorbent assays (ELISA) and polymerase chain reaction (PCR)) offer high sensitivity, but are often limited by complex procedures, time-consuming processes, expensive equipment, and reliance on specialized personnel, making them difficult to meet the aforementioned widespread needs. Optical biosensors, especially those based on luminescence principles, demonstrate great potential in high-sensitivity and high-throughput detection due to their advantages such as high sensitivity, resistance to electromagnetic interference, ease of miniaturization, and visual readout. Examples include: Chemiluminescence (CL): Excites reporter molecules to emit light through chemical reactions (such as luminol and acridinium esters), requiring no external excitation source, exhibiting extremely low background, and high sensitivity. Electrochemiluminescence (ECL): Generates excited-state molecules and emits light through electrochemical reactions on the electrode surface (such as the terpyridine-ruthenium system), combining the control advantages of electrochemistry with the high sensitivity of optical detection, enabling spatiotemporal resolution. Fluorescence (FL): Generates emission light under specific wavelength light excitation (such as quantum dots, fluorescent dyes, and upconversion nanoparticles). Requires an excitation source and filtering system, and also offers relatively high sensitivity.

[0044] In recent years, light-emitting array biosensors have become a cutting-edge solution for addressing the challenges of high-sensitivity multi-channel detection. However, the extraction of light intensity from images obtained using light-emitting array biosensors also suffers from low efficiency and low accuracy.

[0045] Based on this, the present invention provides a method and apparatus for determining the luminescence intensity of an image from an electrochemiluminescence array sensor. The method analyzes and processes mobile phone images from the luminescence array sensor using a Matlab coding platform. Through the graphical user interface module functions in Matlab, the image can be autonomously selected. The image is then rotated, the luminescence region is determined, and the luminescence intensity is automatically extracted. The extraction of the luminescence intensity region is judged through visualization. Finally, the accurately extracted luminescence intensity is saved to an Excel spreadsheet of type "*.xls".

[0046] The technical solutions provided by the various embodiments of the present invention will be described in detail below with reference to the accompanying drawings.

[0047] Figure 1 This is a schematic flowchart of a method for determining the luminescence intensity of an image from an electrochemiluminescence array sensor according to the present invention, which specifically includes the following steps:

[0048] S101: Acquire the terminal imaging image of the electrochemiluminescence array sensor, and remove the background image from the terminal imaging image to obtain the sample image.

[0049] First, the "uigetfile" function in the MATLAB graphical user interface module can be used to visualize and select the terminal imaging image. Enter the command "Main_Function" in the MATLAB command window or open the "Main_Function.m" script file and click "Run" to start the program. The visualization interface will then appear. Select both the background image "bg-line1" and the sample image "line1" to read the terminal imaging image.

[0050] After the background image and sample image are read into the program, the "substractbackground.m" script file is used to generate a numerical matrix by subtracting the sample image and background image after importing them into the program, thereby subtracting the background image from the terminal imaging image.

[0051] S102, with the center of the sample image as the rotation point, rotate the sample image by a preset rotation angle so that the luminous area of ​​the sample image is parallel or perpendicular to the horizontal direction.

[0052] In one embodiment, rotating the sample image by a preset rotation angle with the center of the sample image as the rotation point includes: determining the center of the rotated sample image based on the center of the sample image and the rotation angle; and assigning the new pixel points generated by the rotation to 0 based on the center of the sample image and the center of the rotated sample image to obtain the rotated sample image.

[0053] The rotation angle is determined in advance by adjusting the historical terminal imaging images of the electrochemiluminescence array sensor; the direction of rotation of the sample image is determined by the sign of the rotation angle, with a positive value indicating clockwise rotation and a negative value indicating counterclockwise rotation.

[0054] Specifically, the sample image obtained by removing the background is as follows: Figure 2 As shown in Figure (a), the tilt of the original image's luminous region makes it easy to be interfered with by other regions when selecting the luminous region, making accurate selection of the luminous region's position very difficult. Therefore, the "raotateimge.m" script file is used to calculate the center of the image. Based on this, the center of the new image after rotation and the position of the corresponding points are calculated according to the rotation angle, and then the rotation is performed as follows. Figure 2 As shown in Figure (b), Figure 2 Figure (b) shows a schematic diagram of the selection of the luminous region. During image rotation, some pixels may not correspond properly before and after rotation, resulting in extra pixels. Traditional Matlab image rotation processing mostly relies on the "imrotate" function, which interpolates these extra pixels, leading to differences in light intensity before and after rotation. This causes distortion of the actual luminous intensity, resulting in image enlargement, etc. (The rotated sample image is shown in Figure b). Figure 2As shown in Figure (c) of the original image, this invention proposes a novel image rotation method: The center of the numerical matrix of the sample image is found through calculation. Then, the center of the numerical matrix of the new image after rotation is calculated according to the required rotation angle. Based on the matrix points corresponding to the two centers, any extra pixels that cannot correspond to the new pixels are uniformly assigned a value of 0. This ensures that the intensity of the luminous region remains unchanged before and after rotation. Furthermore, based on the comparison of bright-field and dark-field imaging images, the position of the luminous region is recorded, and the luminous region after rotation is associated and encoded, enabling it to correspond with the original image and the detection image, thus quickly locating each detected hole. Finally, the luminous region of each detection hole is manually determined on the rotated image, and the key coordinates of the luminous region are marked (e.g., ...). Figure 2 The four vertices of the red box in Figure (b) are recorded in the "Main_Function.m" script file. Since the detection strategy and the size and layout of the luminous area in the photograph taken by the same imaging instrument are basically the same, with only slight changes in the position of the luminous area on the image, only minor adjustments are needed in subsequent image processing. Figure 2 The coordinates of the red box in Figure (b) are used to accurately select the luminous area, which takes about 5-20 seconds, greatly improving image processing efficiency and saving analysis time.

[0055] S103, extract the coordinate data of the luminous region based on the rotated sample image.

[0056] In one embodiment, extracting coordinate data of the luminous region based on the rotated sample image includes: determining the luminous region of the rotated sample image through a human-computer interaction interface; determining whether the luminous region is selected correctly; and extracting the coordinate data of the luminous region if the luminous region is selected correctly.

[0057] Optionally, determining whether the luminous region is selected correctly includes: obtaining the grayscale values ​​of all pixels on the boundary line of the luminous region, the average grayscale value of the non-luminous region, and three times the standard deviation of the non-luminous region; if the grayscale values ​​of all pixels on the boundary line of the luminous region are less than the sum of the average grayscale value of the non-luminous region and three times the standard deviation, then the luminous region is determined to be selected correctly; otherwise, the luminous region is selected incorrectly.

[0058] In cases where the luminous region is selected incorrectly, the luminous region of the rotated sample image is reselected until the correct luminous region is selected.

[0059] Specifically, the accuracy of the selection of the luminous area is generally determined by visual observation. Figure 2Whether the red box in Figure (b) can completely enclose the luminous area depends heavily on the analyst's subjective judgment, potentially leading to inaccurate analysis. Therefore, to determine if the red box completely covers all luminous areas, the script file "croparea.m" calculates the grayscale value of all pixels along the red box line, comparing it to the average and standard deviation of the grayscale values ​​in the non-luminous area. Only when the average grayscale value of all pixels along the red box line in every luminous area is less than the average of the non-luminous area plus three times the standard deviation is the selected area considered accurate; otherwise, an error dialog box will pop up. Figure 3 As shown, if a portion of the illuminated area is found to be outside the red frame during calculation, a message will appear: "Selection area mismatch". The coordinates of the red frame need to be readjusted by changing the coordinate values ​​of the red frame.

[0060] S104. Based on the coordinate data of the luminous area, determine the gray value of the luminous area, and determine the total gray value of all luminous areas as the luminous intensity of the terminal imaging image.

[0061] After the light intensity of the luminous region is accurately extracted, the extracted values ​​can be visualized and written to an Excel file using the "xlswrite" function in Matlab.

[0062] In one embodiment, such as Figure 4 As shown, Figure 4 This is a flowchart illustrating the implementation of the method for determining the luminescence intensity of an electrochemiluminescence array sensor image provided by the present invention. The present invention mainly analyzes and processes mobile phone imaging images of the luminescence array sensor based on the Matlab coding platform. Through the graphical user interface module functions in Matlab, the imaging image can be autonomously selected, and further the imaging image is rotated, the luminescence region is determined, the luminescence intensity is automatically extracted, and the extraction of the luminescence intensity region of the imaging image is judged through visualization. Finally, the accurately extracted luminescence intensity is saved to an Excel spreadsheet of type "*.xls".

[0063] The Matlab scripts used to extract luminescence intensity from imaging images mainly consist of four files: "Main_Function.m", "substractbackground.m", "raotateimge.m", and "croparea.m". The "Main_Function.m" script is the main program, which analyzes and processes the imaging image by coding and calling the other three scripts. The "substractbackground.m" script function reads the background and sample images into the program to generate matrix data, then subtracts the background matrix data from the sample matrix data to obtain a new matrix data, thus achieving background subtraction. The "raotateimge.m" script function finds the center of the matrix for background subtraction, then rotates the matrix data by a certain angle so that the tilted luminescence area is parallel or perpendicular to the image direction. The rotation angle is determined through debugging during the initial run, and the direction of rotation can be determined by the sign (positive or negative) and is set in the "Main_Function.m" script file, thereby achieving the rotation of the imaging image. The "croparea.m" script function uses the manually determined coordinates of the luminous areas from the rotated image, extracts the matrix data after removing the background, and calculates the total grayscale intensity of each selected area, which is the total luminous intensity. To start this program, simply enter the command "Main_Function" in the Matlab command window or open the "Main_Function.m" script file and click "Run".

[0064] When applying the method for determining the luminescence intensity of an electrochemiluminescence array sensor image provided by this invention, it is not necessary to rely on... Figure 1 The steps shown are executed in sequence. The specific execution order of each step can be determined as needed, and this invention does not impose any restrictions on it.

[0065] The above describes a method for determining the luminescence intensity of an electrochemiluminescence array sensor image according to one or more embodiments of the present invention. Based on the same idea, the present invention also provides a corresponding apparatus for determining the luminescence intensity of an electrochemiluminescence array sensor image, the apparatus comprising:

[0066] The acquisition module is used to acquire the terminal imaging image of the electrochemiluminescence array sensor and remove the background image from the terminal imaging image to obtain the sample image;

[0067] The rotation module is used to rotate the sample image by a preset rotation angle with the center of the sample image as the rotation point, so that the luminous area of ​​the sample image is parallel or perpendicular to the horizontal direction.

[0068] The extraction module is used to extract the coordinate data of the luminescent region based on the rotated sample image;

[0069] The determination module is used to determine the grayscale value of the luminous area based on the coordinate data of the luminous area, and to determine the total grayscale value of all luminous areas as the luminous intensity of the terminal imaging image.

[0070] Specific limitations regarding the device for determining the luminescence intensity of an electrochemiluminescence array sensor image can be found in the limitations of the method for determining the luminescence intensity of an electrochemiluminescence array sensor image described above, and will not be repeated here. Each module in the aforementioned device for determining the luminescence intensity of an electrochemiluminescence array sensor image can be implemented entirely or partially through software, hardware, or a combination thereof. These modules can be embedded in or independent of the processor in a computer device, or stored in the memory of a computer device as software, so that the processor can call and execute the corresponding operations of each module.

[0071] The present invention also provides a computer-readable storage medium storing a computer program that can be used to execute the above-described... Figure 1 A method for determining the luminescence intensity of an electrochemiluminescence array sensor image is provided.

[0072] The present invention also provides Figure 5 The schematic diagram of the computer device shown is as follows: Figure 5 As shown, at the hardware level, this computer device includes a processor, internal bus, network interface, memory, and non-volatile memory, and may also include other hardware required for business operations. The processor reads the corresponding computer program from the non-volatile memory into memory and then executes it to achieve the above. Figure 1 A method for determining the luminescence intensity of an electrochemiluminescence array sensor image is provided.

[0073] Those skilled in the art will understand that all or part of the processes in the methods of the above embodiments can be implemented by a computer program instructing related hardware. The computer program can be stored in a non-volatile computer-readable storage medium, and when executed, it can include the processes of the embodiments of the methods described above. Any references to memory, storage, databases, or other media used in the embodiments provided by this invention can include at least one of non-volatile and volatile memory. Non-volatile memory can include read-only memory (ROM), magnetic tape, floppy disk, flash memory, or optical storage, etc. Volatile memory can include random access memory (RAM) or external cache memory. By way of illustration and not limitation, RAM can be in various forms, such as static random access memory (SRAM) or dynamic random access memory (DRAM), etc.

[0074] The technical features of the above embodiments can be combined in any way. For the sake of brevity, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this invention.

Claims

1. A method for determining the luminescence intensity of an image from an electrochemiluminescence array sensor, characterized in that, include: Acquire the terminal imaging image of the electrochemiluminescence array sensor, and remove the background image from the terminal imaging image to obtain the sample image; Using the center of the sample image as the rotation point, rotate the sample image by a preset rotation angle so that the luminous area of ​​the sample image is parallel or perpendicular to the horizontal direction; Based on the rotated sample image, the luminous region of the rotated sample image is determined through the human-computer interaction interface; it is determined whether the luminous region is selected correctly, and if the luminous region is selected correctly, the coordinate data of the luminous region is extracted; Based on the coordinate data of the luminous areas, the grayscale values ​​of the luminous areas are determined, and the total grayscale values ​​of all luminous areas are used to determine the luminous intensity of the terminal imaging image; The process of determining whether the luminous region has been selected correctly includes: obtaining the gray values ​​of all pixels on the boundary line of the luminous region, the average gray value of the non-luminous region, and three times the standard deviation of the non-luminous region; if the gray values ​​of all pixels on the boundary line of the luminous region are less than the sum of the average gray value of the non-luminous region and three times the standard deviation, then the luminous region has been selected correctly. Otherwise, if the luminous region is selected incorrectly, the luminous region of the rotated sample image will be reselected until the luminous region is selected correctly.

2. The method according to claim 1, characterized in that, Rotate the sample image by a preset rotation angle, using the center of the sample image as the rotation point, including: Determine the center of the rotated sample image based on the center of the sample image and the rotation angle; Based on the center of the sample image and the center of the rotated sample image, the new pixels generated by the rotation are assigned a value of 0 to obtain the rotated sample image.

3. The method according to claim 1, characterized in that, The rotation angle was determined in advance by adjusting historical terminal imaging images of the electrochemiluminescence array sensor.

4. The method according to claim 1, characterized in that, The direction of rotation of the sample image is determined by the sign of the rotation angle.

5. A device for determining the luminescence intensity of an image from an electrochemiluminescence array sensor, characterized in that, The apparatus is used to implement the method according to any one of claims 1-4, the apparatus comprising: The acquisition module is used to acquire the terminal imaging image of the electrochemiluminescence array sensor and remove the background image from the terminal imaging image to obtain the sample image; The rotation module is used to rotate the sample image by a preset rotation angle with the center of the sample image as the rotation point, so that the luminous area of ​​the sample image is parallel or perpendicular to the horizontal direction. The extraction module is used to extract the coordinate data of the luminescent region based on the rotated sample image; The determination module is used to determine the grayscale value of the luminous area based on the coordinate data of the luminous area, and to determine the total grayscale value of all luminous areas as the luminous intensity of the terminal imaging image.

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