WO2020037572A1 - Procédé et dispositif de détection d'un point lumineux sur une image, et procédé et dispositif d'enregistrement d'image - Google Patents

Procédé et dispositif de détection d'un point lumineux sur une image, et procédé et dispositif d'enregistrement d'image Download PDF

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WO2020037572A1
WO2020037572A1 PCT/CN2018/101817 CN2018101817W WO2020037572A1 WO 2020037572 A1 WO2020037572 A1 WO 2020037572A1 CN 2018101817 W CN2018101817 W CN 2018101817W WO 2020037572 A1 WO2020037572 A1 WO 2020037572A1
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image
bright spot
candidate
registration
bright
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PCT/CN2018/101817
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English (en)
Chinese (zh)
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李林森
徐伟彬
金欢
姜泽飞
周志良
颜钦
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深圳市真迈生物科技有限公司
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Priority to PCT/CN2018/101817 priority Critical patent/WO2020037572A1/fr
Publication of WO2020037572A1 publication Critical patent/WO2020037572A1/fr

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    • G—PHYSICS
    • G06—COMPUTING; CALCULATING OR COUNTING
    • G06T—IMAGE DATA PROCESSING OR GENERATION, IN GENERAL
    • G06T7/00—Image analysis

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  • the invention relates to the field of image processing, in particular to a method for detecting bright spots on an image, a device for detecting bright spots on an image, a computer program product containing instructions for detecting bright spots on an image, and an image Registration method, an image registration device, and a computer program product containing instructions for image registration.
  • image analysis is a very important part. It depends on the detection and recognition of bright spots on the image and the detection and recognition of bright spots into bases / nucleosides. Acid sequence to achieve nucleic acid sequence determination. The accuracy of detection and localization of bright spots on the image directly determines the accuracy of gene sequencing.
  • nucleic acid sequence determination In the application of nucleic acid sequence determination, how to detect bright spots on an image simply and quickly and / or effectively and use the bright spot information for image processing needs to be developed and improved.
  • the embodiments of the present invention aim to solve at least one of the technical problems in the related art or provide at least one optional practical solution.
  • a method for detecting bright spots on an image is provided.
  • the so-called image is collected from a field where a base extension reaction occurs, and there are multiple bands on the field where the base extension reaction occurs.
  • the method includes: using a k1 * k2 matrix to detect bright spots on the image, and determining whether the central pixel value of the matrix is not.
  • a matrix that is smaller than the first preset value, that the non-central pixel value of the matrix is not smaller than the second preset value, and that the central pixel value of the matrix is not smaller than any pixel value that is not the center of the matrix corresponds to a candidate bright spot, k1 and k2 Both are odd numbers greater than 1.
  • the k1 * k2 matrix contains k1 * k2 pixels.
  • the so-called first preset value and / or the second preset value are related to the average pixel value of the image; and the so-called candidate brightness is determined. Whether the spot is bright.
  • an image registration method which includes using bright spots on an image for registration, and the light spots on the image are determined using the bright spot detection method in the embodiment of the present invention.
  • the registration method includes: performing a first registration on a to-be-registered image based on a reference image, the reference image and the to-be-registered image corresponding to the same field of view, including determining a position on the to-be-registered image.
  • a first offset of a predetermined region and a corresponding predetermined region on the reference image, all bright spots on the image to be registered are moved based on the first offset, to obtain a first registration to be registered Image; performing second registration on the image to be registered after the first registration based on the reference image, including merging the image to be registered after the first registration and the reference image, obtaining a merged image, calculating The offsets of all overlapping bright spots in a predetermined area on the merged image are determined to determine a second offset, and two or more bright spots with a distance less than a predetermined pixel are one of the overlapping bright spots, based on the second offset
  • the shift amount moves all bright spots on the image to be registered after the first registration to achieve registration of the image to be registered.
  • a computer-readable storage medium for storing a program for execution by a computer, and executing the program includes performing bright spot detection and / or image configuration on an image in any of the foregoing embodiments. Quasi-method.
  • the computer-readable storage medium may include: a read-only memory, a random access memory, a magnetic disk, or an optical disk.
  • a computer program product includes instructions for implementing detection of bright spots on an image.
  • the instructions When the computer executes the program, the instructions cause the computer to execute the bright spots in the embodiment of the present invention. Some or all steps of the detection method.
  • a computer program product includes instructions for implementing image registration.
  • the instruction causes the computer to execute a part of the image registration method in the embodiment of the present invention or All steps.
  • the so-called “bright spot” or “bright spot” refers to a light emitting point on an image, and one light emitting point occupies at least one pixel point.
  • the so-called “pixel” is the same as “pixel”.
  • the image comes from a sequencing platform that uses optical imaging principles for sequence determination.
  • the so-called sequencing platform includes, but is not limited to, BGI, Illumina / Solexa, ThermoFisher / Life Technologies / ABI, SOLiD, Roche 454, and PacBio
  • the method, device and system / computer program product for detecting bright spots on an image provided by any one of the above embodiments of the present invention, it is possible to quickly and effectively detect bright spots on an image, especially for a sequence collected from a nucleic acid sequence.
  • the image of the reaction was measured.
  • the method has no special restrictions on the detection image, ie, the original input data, and is applicable to the processing and analysis of images generated by any platform that uses the principle of optical detection for nucleic acid sequence determination, including image quality assessment for focus and focus, including alkali Based on the image processing and analysis of recognition, it has the characteristics of high accuracy and efficiency, and can obtain more information representing the sequence from the image.
  • second-generation sequencing generally contains nucleic acid templates.
  • Signal amplification (such as amplification).
  • a nucleic acid template exists in the form of a cluster containing at least hundreds or thousands of copies. That is, the signal of the nucleic acid template is a large number of signal sets of the nucleic acid template molecule.
  • the signal reflected on the image is strong and / or has specific morphological characteristics. It can also be said that the signal is significantly different from the non-target signal, and it is relatively easy to identify and locate. Therefore, the detection of bright spots on general second-generation sequencing images does not require special image processing and does not require a comprehensive and highly accurate identification of the bright spots of the corresponding sequence information. A large number of bright spots of the corresponding sequence can be obtained. Spot signals are then identified and converted into bright spot signals as sequence information.
  • the sequencing chip used is random, that is, the probes on the sequencing chip are randomly arranged, and the images obtained by taking pictures are random ( random) image, which is not easy to process and analyze; moreover, in general single-molecule sequencing, because the method does not include a nucleic acid template, the nucleic acid template exists as a single molecule or a few molecules, which is reflected in the image and is weak and easy to be disturbed / submerged
  • the accurate identification of the bright spots corresponding to the nucleic acid molecule and the amount of bright spots identified directly determine the throughput and the amount of effective data.
  • single-molecule sequencing has high requirements for image processing and bright spot positioning. It is hoped that all effective bright spots on the image can be identified and accurately located so that as much accurate data as possible can be obtained.
  • single molecule is meant one or a few molecules, such as no more than 10 molecules.
  • the method, device and / or corresponding computer product for detecting bright spots on an image according to the embodiments of the present invention are suitable for detecting bright spots on a sequenced image, especially for random images and signal recognition with high accuracy requirements, especially Advantages.
  • the image registration method, device, and / or terminal product according to the embodiments of the present invention include registration / correction of an image based on bright spots on the image, and can achieve high-precision correction with a smaller amount of data, which is particularly applicable.
  • image information required for high accuracy / high precision such as correction of images from sequencing platforms.
  • FIG. 1 is a schematic flowchart of a method for detecting bright spots on an image in a specific embodiment of the present invention.
  • FIG. 2 is a schematic diagram of a matrix and connected pixels corresponding to candidate bright spots in an image in an embodiment of the present invention.
  • FIG. 3 is a schematic flowchart of an image registration method in a specific embodiment of the present invention.
  • FIG. 4 is a schematic diagram of a correction process and a correction result in a specific embodiment of the present invention.
  • FIG. 5 is a schematic structural diagram of a device for detecting bright spots on an image in a specific embodiment of the present invention.
  • FIG. 6 is a schematic structural diagram of an image registration device in a specific embodiment of the present invention.
  • the sequencing in the embodiment of the present invention is also referred to as sequence determination, and refers to nucleic acid sequence determination, including DNA sequencing and / or RNA sequencing, including long-sequence sequencing and / or short-sequence sequencing.
  • Sequencing can be performed through a sequencing platform.
  • the sequencing platform can be selected but not limited to Hisq / Miseq / Nextseq sequencing platform from Illumina, IonTorrent platform from Thermo Fisher / Life Technologies, BGISEQ platform and single molecule sequencing platform from BGI; sequencing method You can choose single-end sequencing or double-end sequencing; the obtained sequencing results / data are the read out fragments, which are called reads. The length of the read segment is called the read length.
  • a method for detecting bright spots on an image is from a field where a base extension reaction occurs, and there are multiple bands on the field where the base extension reaction occurs.
  • the method includes: S10 uses a k1 * k2 matrix to detect bright spots on the image, and determines that the center pixel value of the matrix is not less than the first A matrix with a preset value, a non-central pixel value of the matrix not less than the second preset value, and a central pixel value of the matrix not less than any non-center pixel value of the matrix corresponds to a candidate bright spot, k1 and k2 are both odd numbers greater than 1.
  • K1 * k2 matrix contains k1 * k2 pixels; and S20 determines whether the candidate bright spot is a bright spot.
  • the above method for detecting bright spots on an image can quickly and effectively detect bright spots on an image, especially an image collected from a nucleic acid sequence measurement response.
  • the method has no special restrictions on the detection images, ie, the original input data, and is applicable to the processing and analysis of images generated by any platform that uses the principle of optical detection for nucleic acid sequence determination, including but not limited to second- and third-generation sequencing. Efficient feature, can get more representative sequence information from the image. It is especially advantageous for signal recognition with random images and high accuracy requirements.
  • the image comes from a nucleic acid sequence determination reaction.
  • the nucleic acid molecule is provided with an optically detectable label, such as a fluorescent label.
  • the fluorescent molecule can be excited to emit fluorescence under laser irradiation at a specific wavelength, and the image is acquired by an imaging system.
  • the acquired images include light spots / bright spots that may correspond to the location of the fluorescent molecules. Understandably, when in the focal position, the size of the bright spot corresponding to the position of the fluorescent molecule in the collected image is small and the brightness is high; when it is in the non-focus position, the collected image The size of the bright spot corresponding to the position of the fluorescent molecule is larger and the brightness is lower.
  • the so-called single molecule is a few molecules, for example, the number of molecules is not more than 10, for example, one, two, three, four, five, six, eight or ten.
  • S10 may use a k1 * k2 matrix to perform ergodic recognition and detection of bright spots on an image.
  • the pixel values are the same as the grayscale values.
  • k1 * k2 matrix, k1 and k2 may be equal or unequal.
  • the relevant parameters of the imaging system are: the objective lens is 60 times, the size of the electronic sensor is 6.5 ⁇ m, and the image formed by the microscope and then passed through the electronic sensor, the minimum size that can be seen is 0.1 ⁇ m.
  • It can be a 16-bit grayscale or color image of 512 * 512, 1024 * 1024, or 2048 * 2048.
  • the inventor has performed a large number of image processing statistics, taking the first preset value as 1.4 times the average pixel value of the image, and taking the second preset value as 1.1 times the average pixel value of the image, which can eliminate interference, Spot detection results obtained from optical detection marks.
  • the size, similarity and / or intensity of the ideal bright spot can be used to further screen and judge candidate bright spots.
  • the size of the candidate bright spots on the comparison image is quantitatively reflected by using the size of the connected domain corresponding to the candidate bright spots, so as to filter and determine whether the candidate bright spots are the desired bright spots.
  • the average pixel value of the image is used as a reference, and two or more adjacent pixels that are not less than the average pixel value are called connected pixels / connectivity, as shown in FIG. 2, Bold and enlarged represents the center of the matrix corresponding to the bright spot, and the thick line frame represents the 3 * 3 matrix corresponding to the candidate bright spot.
  • the so-called third preset value may be determined according to the information of the size of the connected domain corresponding to all candidate bright spots on the image. For example, by calculating the size of the connected domain corresponding to each candidate bright spot on the graph, taking the average value of the size of the connected domain of the bright spots represents a characteristic of the image as a third preset value; for example, each candidate in the image may be The size of the connected domain corresponding to the bright spot is sorted from small to large, and the size of the connected domain at the 50th, 60th, 70th, 80th, or 90th quantile is taken as the third preset value. In this way, the bright spot information can be effectively obtained, which is beneficial for subsequent recognition of the nucleic acid sequence.
  • statistically set parameters are used to quantitatively reflect the intensity characteristics of the comparison candidate bright spots, thereby filtering the candidate bright spots.
  • the so-called fourth preset value may be determined according to the information of the magnitudes of the scores of all candidate bright spots on the image. For example, when the number of candidate bright spots on the image is greater than a certain number, which meets the statistical requirements, for example, the number of candidate bright spots on the image is greater than 30, the score values of all candidate bright spots on the image can be calculated and Ascending order, the fourth preset value can be set to the 50th, 60th, 70th, 80th, or 90th quantile Score value, so that less than 50th, 60th, 70th, 80th, or 90th can be excluded.
  • the candidate bright spots of the quantile Score value are helpful for effectively obtaining the target bright spots and accurate subsequent recognition of the base sequence.
  • the basis for performing this processing or the screening setting is that, generally, it is considered that the bright spots that have a large difference in intensity and pixel value between the center and the edge and are converged are bright spots corresponding to the location of the molecule to be detected.
  • the number of candidate bright spots on the image is greater than 50, greater than 100, or greater than 1,000.
  • candidate bright spots are screened based on morphology and intensity / brightness.
  • a connected pixel that is larger than the average pixel value in a k1 * k2 matrix as a connected domain corresponding to a candidate bright spot.
  • the center pixel value of the matrix corresponding to the spot, EV represents the sum of the non-center pixel values of the matrix corresponding to the bright spot; the candidate bright spot whose size of the corresponding connected domain is larger than the third preset value and the score is greater than the fourth preset value is determined For a bright spot.
  • the so-called third preset value and / or fourth preset value may be considered and set with reference to the foregoing specific implementation manner.
  • the image is a pre-processed image
  • the pre-processing includes removing the background and / or the Mexican hat transform.
  • the manner or tool method used to implement the background removal and / or Mexican hat transformation there is no limitation on the manner or tool method used to implement the background removal and / or Mexican hat transformation.
  • de-backgrounding an image includes performing an open operation to estimate the background of the image, and then subtracting the background from the original image to obtain the de-backgrounded image. The open operation is used to eliminate small objects, separate objects at slim points, and smooth the boundaries of larger objects without changing the image area significantly, which is beneficial for subsequent processing and analysis of the image.
  • a window / matrix of a predetermined size is set to perform Gaussian filtering on the image before filtering, and two-dimensional Laplacian sharpening is performed on the Gaussian filtered image.
  • Mexican hat filtering is implemented in two steps.
  • the image pre-processing includes removing the background, and estimating the background of the image based on the mean gray value of at least a part of the region of the image. In this way, it is easier and more effective to retain and highlight the target information on the image.
  • the image is a binarized image.
  • reference may be made to the binarization method described in CN107945150A and CN107918931A.
  • a result obtained by using a Mexican hat filter of an image based on a pixel matrix of a predetermined size is divided by a result of an image opening operation to obtain a set of noise corresponding to the pixel matrix of a predetermined size, and a threshold is set based on the set of noise.
  • the pixel matrix of a predetermined size is 15 * 15. This is good for binarization threshold calculation and realization of bright spot detection.
  • the image is a binary image, and it is determined that the candidate bright spot whose size of the corresponding connected pixel is not less than a fifth preset value is one bright spot.
  • the connected pixels are also called connected pixels.
  • the fifth preset value is not less than two-thirds of the matrix. In this way, the bright spot information corresponding to the nucleic acid molecule and convenient for subsequent rapid analysis can be effectively obtained.
  • the method further includes: if it is determined that the candidate bright spot is a bright spot, calculating the intensity value of the sub-pixel center coordinate and / or the sub-pixel center coordinate of the bright spot, and if it is determined that the candidate bright spot is not a bright spot; , Discard candidate bright spots.
  • calculating the sub-pixel center coordinates of the bright spot and / or the intensity value of the sub-pixel center coordinates includes: using a quadratic function interpolation to calculate the sub-pixel center coordinates of the bright spot, and / or using a quadratic spline interpolation to calculate the sub-pixel center coordinates. The intensity value of the pixel center coordinates. In this way, the method using a quadratic function and / or a quadratic spline can further improve the accuracy of judging the bright spots of the image and locating the bright spots.
  • An embodiment of the present invention further provides an image registration method, which includes using bright spots on an image to achieve registration.
  • the bright spots on the image are obtained by using the method for detecting bright spots on an image in any of the foregoing specific implementation manners.
  • the description of the advantages and technical features of the method for detecting bright spots on an image in any of the foregoing specific implementation manners is also applicable to the image registration method in this embodiment manner, and details are not described herein again.
  • the so-called image registration method method includes: S100 performs first registration based on a reference image to be registered, and the reference image and the image to be registered correspond to the same field of view, including, determining A first offset of a predetermined region on the image to be registered and a corresponding predetermined region on the reference image, all bright spots on the image to be registered are moved based on the first offset to obtain a first registration to be registered Image; S200 performs second registration on the image to be registered after the first registration based on the reference image, including merging the image to be registered and the reference image after the first registration, obtaining a merged image, and calculating a reservation on the merged image The offset of all overlapping bright spots in the area to determine a second offset.
  • Two or more bright spots with a distance less than a predetermined pixel are one overlapping bright spot.
  • All bright spots on the image to be registered to achieve registration of the image to be registered.
  • coarse registration coarse correction
  • fine registration fine correction
  • the predetermined area on the so-called image may be the entire image or at least a part of the image.
  • the predetermined area on the image is a part of the image, such as a 512 * 512 area in the center of the image.
  • the so-called image center is the center of the field of view.
  • the intersection between the optical axis of the imaging system and the imaging plane can be referred to as the image center point, and the area centered on the center point can be regarded as the image center area.
  • the image to be registered comes from a nucleic acid sequencing platform
  • the platform includes an imaging system and a nucleic acid sample carrying system
  • the nucleic acid molecule to be tested with an optical detection label is fixed in a reactor, which is also called
  • the chip is mounted on a movable table, and the moving table drives the chip to realize image acquisition of the nucleic acid molecules to be tested located at different positions (different fields of view) of the chip.
  • there is a limit on the accuracy of the movement of the optical system and / or the mobile stage For example, there is a deviation between the specified movement to a certain position and the position reached by the actual movement of the mechanical structure, especially in application scenarios that require high accuracy.
  • the so-called reference image is obtained by construction.
  • the construction of the so-called reference image includes: acquiring a first image and a second image, the first image and the second image corresponding to the same field of view as the image to be registered; performing coarse registration of the second image based on the first image, including determining the second image An offset between the image and the first image, and the second image is moved based on the offset to obtain a second image after the coarse registration; the first image and the second image after the coarse registration are combined to obtain a reference image.
  • the use of multiple images to construct a reference image facilitates the reference image to obtain complete bright spot information of the corresponding nucleic acid molecule and facilitates correction of the bright spot-based image.
  • the reference image and the image to be registered are binarized images. In this way, it is beneficial to reduce the amount of calculation and quickly correct the deviation.
  • a two-dimensional discrete Fourier transform is used to determine the first offset and / or the second image and the first image. In this way, the first offset amount and / or the second image and the offset amount of the first image can be determined quickly and accurately.
  • the image to be corrected and the reference image are both binary images, that is, each pixel in the image is not a or b, for example, a is 1, b is 0, and a pixel labeled 1 is brighter than a pixel labeled b. , Or high intensity; during nucleic acid sequencing, defining a nucleic acid molecule to be extended by one base or one base is called a cycle, and the reference image is constructed using the images cyclic1-cycle4 of cycles 1-4, The first image and the second image are selected from any one, two, or three of the images cycle1-cycle4. In one example, the first image is the image cycle1, and the image cycle2-4 is the second image.
  • the images cycle2-4 are sequentially coarsely registered to obtain the coarsely registered images cycle2-4, respectively; the merged images
  • a reference image was obtained from cycle1 and the coarsely registered image cycle2-4.
  • the so-called merged image is an overlapping bright spot in the merged image. It is mainly based on the size of the bright spot of the corresponding nucleic acid molecule and the resolution of the imaging system. In one example, two bright spots with a distance of no more than 1.5 pixels on the two images are set as coincident bright spots.
  • the composite image center area of 4 cycles is used as the reference image, which is helpful to make the reference image have a sufficient amount of bright spots and facilitate subsequent registration, and secondly, to detect and locate the bright spots in the central area of the image.
  • the speckle information is relatively more accurate and facilitates accurate registration.
  • the following steps are performed to correct the image: 1) Rough correction is performed on the cycle5 image of a certain field of vision collected from the fifth round of response.
  • Cycle5 is a binary image, and the center of the image is taken as 512 * 512.
  • two bright spots with a distance of no more than 1.5 pixels on the two images are set as coincident bright spots; 3)
  • a field-of-view image (fov) with offsets (x0, y0) of different cycles is obtained.
  • -(x1, y1), for a bright spot (peak) can be expressed as: curCyclePoints + (x0, y0)-(x1, y1)
  • curCyclePoints represents the original coordinates of the bright spot, that is, the coordinates in the image before correction.
  • the correction result obtained by the above image correction has higher accuracy, and the correction accuracy is less than or equal to 0.1 pixels.
  • Figure 4 shows the correction process and results.
  • image C is corrected based on image A.
  • the circles in image A and image C indicate bright spots.
  • Bright spots with the same digital mark are coincident bright spots.
  • Image C-> A indicates The correction result, that is, the image C is aligned to the image A.
  • a "computer-readable storage medium” may be any device that can contain, store, communicate, propagate, or transmit a program for use by or in connection with an instruction execution system, apparatus, or device. .
  • computer-readable storage media include the following: electrical connections (electronic devices) with one or more wirings, portable computer disk cartridges (magnetic devices), random access memory (RAM) , Read-only memory (ROM), erasable and editable read-only memory (EPROM or flash memory), fiber optic devices, and portable optical disk read-only memory (CDROM).
  • the computer-readable storage medium may even be paper or other suitable media on which the program can be printed, because, for example, by optically scanning the paper or other media and then editing, interpreting, or otherwise Processing is performed in a suitable manner to obtain the program electronically and then store it in a computer memory.
  • an embodiment of the present invention provides a device 1000 for detecting bright spots on an image.
  • the device is configured to implement the method for detecting bright spots on an image in any of the foregoing specific implementation manners.
  • a field of view where a base extension reaction occurs.
  • the device includes: candidate bright spots A judging module 1010, which is a candidate bright spot judging module for detecting a bright spot on an image by using a k1 * k2 matrix.
  • a matrix with a preset value and the center pixel value of the matrix is not less than any non-center pixel value of the matrix corresponds to a candidate bright spot
  • k1 and k2 are odd numbers greater than 1
  • the k1 * k2 matrix contains k1 * k2 pixels
  • the first The preset value and / or the second preset value are related to an average pixel value of the image
  • a bright spot determination template 1020 is used to determine whether the candidate bright spot from the candidate bright spot determination module is a bright spot.
  • A represents the connected size of the row with the center of the matrix corresponding to the candidate bright spot
  • B represents the connected size of the column with the center of the matrix corresponding to the candidate bright spot, and defines connected pixels that are larger than the average pixel value as a connected domain
  • / Or calculate the score of a candidate bright spot Score ((k1 * k2-1) CV-EV) / ((CV + EV) / (k1 * k2)), determine the candidate whose score is greater than the fourth preset value
  • the bright spot is a bright spot
  • CV represents the central pixel value of the matrix corresponding to the candidate bright spot
  • EV represents the sum of the non-center pixel values of the matrix corresponding to the bright spot.
  • the bright spot determination template 1020 is used to determine that the candidate bright spot whose size of the corresponding connected pixel is not less than a fifth preset value is one bright spot.
  • the apparatus 1000 further includes an image pre-processing module 1030.
  • the image pre-processing module 1030 is configured to perform background removal and / or Mexican hat transformation on the image. Further, in the candidate bright spot determination module 1010, candidate bright spot detection is performed on an image from the image preprocessing module 1030.
  • background removal is performed based on a gray average value of at least a part of an image.
  • the image is a binarized image.
  • the apparatus 1000 for detecting bright spots on an image further includes a bright spot coordinate determining module 1040, where the coordinate determining module is configured to: if the candidate bright spots are determined to be bright spots in the bright spot determining module, calculate The brightness value of the sub-pixel center coordinate and / or the intensity value of the sub-pixel center coordinate. If it is determined in the bright-spot determination module that the candidate bright spot is not a bright spot, the candidate bright spot is discarded.
  • An embodiment of the present invention further provides an image registration device 2000.
  • the device uses bright spots on an image for registration, and includes the device 1000 for detecting bright spots on an image in any of the foregoing embodiments.
  • the above description of the device for detecting bright spots on an image in any embodiment of the present invention and the advantages and technical features of the image registration method are also applicable to the image registration apparatus 1000 according to the embodiment of the present invention, and details are not described herein again.
  • the image registration device 2000 includes: a first registration module 2010, configured to perform first registration of a to-be-registered image based on a reference image, the reference image, and the The registration image corresponds to the same field of view, including determining a first offset of a predetermined region on the image to be registered and a corresponding predetermined region on the reference image, and moving all bright spots on the image to be registered based on the first offset, Obtaining a first to-be-registered image after registration; a second registration module 2020 for performing second registration on the first-to-be-registered image from the first registration module based on a reference image, including: Combine the to-be-registered image and the reference image after the first registration to obtain a merged image, and calculate the offsets of all overlapping bright spots in a predetermined area on the merged image to determine the second offset.
  • the distance is less than two pixels of the predetermined pixel.
  • One or more bright spots are a coincident bright spot
  • the image registration device 2000 further includes a reference image construction module 2030.
  • the reference image construction module 2030 is configured to construct a reference image, including: acquiring a first image and a second image, and a first image and a second image. Corresponds to the same field of view as the image to be registered; performing coarse registration on the second image based on the first image, including determining an offset between the second image and the first image, and moving the second image based on the offset to obtain a coarse registration The second image after the first image is merged with the second image after the coarse registration to obtain the reference image. Furthermore, the first registration module 2010 performs the first registration according to the reference image 2030 from the reference image construction module.
  • the reference image and the image to be registered are binarized images.
  • a two-dimensional discrete Fourier transform is used to determine the first offset and / or the second image and the first image.
  • Each functional unit / module in each of the foregoing embodiments may be integrated into one processing module, or each unit may exist separately physically, or two or more units may be integrated into one module.
  • the above integrated modules may be implemented in the form of hardware or software functional modules. If the integrated module is implemented in the form of a software functional module and sold or used as an independent product, it may also be stored in a computer-readable storage medium.
  • an embodiment of the present invention also provides a computer program product, the program product includes instructions for implementing detection of bright spots on an image, and the instructions, when the computer executes the program, cause the computer to execute the detection image in any one of the embodiments of the present invention described above Bright spot method.
  • the description of the advantages and technical features of the method for detecting bright spots on an image in any of the foregoing embodiments is also applicable to the computer program product of this embodiment of the present invention, and details are not described herein again.
  • An embodiment of the present invention also provides another computer program product, which includes instructions for implementing image registration.
  • the instruction causes the computer to execute the image registration method in any one of the embodiments of the present invention.
  • the description of the advantages and technical features of the image registration method in any embodiment is also applicable to the computer program product of this embodiment of the present invention, and details are not described herein again.
  • An embodiment of the present invention provides a system for detecting bright spots on an image, including a data input device for inputting data; a data output module for outputting data; a storage device for storing data including a computer-readable program; and
  • the processor is configured to execute a computer-readable program, and the execution of the computer-readable program includes all or part of steps of a method for detecting a bright spot on an image in any of the embodiments of the present invention described above.
  • An embodiment of the present invention provides an image registration system including a data input device for inputting data; a data output module for outputting data; a storage device for storing data including a computer-readable program; and a processor for To execute a computer-readable program, executing the computer-readable program includes all or part of the steps for implementing the image registration method in any one of the embodiments of the present invention described above.
  • controller in addition to implementing the controller / processor in a pure computer-readable program code manner, the controller can be controlled by logic gates, switches, ASICs, and editable logic by logically changing the method steps. Controller and embedded microcontroller to achieve the same function. Therefore, such a controller / processor can be considered as a hardware component, and a device included therein for implementing various functions can also be considered as a structure within the hardware component. Or even, the means for implementing various functions can be regarded as a structure that can be both a software module implementing the method and a hardware component.

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  • Theoretical Computer Science (AREA)
  • Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
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Abstract

Procédé et dispositif (1000) pour détecter un point lumineux sur une image, et procédé et dispositif d'enregistrement d'image (2000). L'image ainsi appelée provient d'un champ de vision où se produit une réaction d'extension de base, et dans le champ de vision où se produit la réaction d'extension de base, il y a une pluralité de molécules d'acide nucléique ayant des marques optiquement détectables, au moins certaines des molécules d'acide nucléique apparaissant sous la forme de points lumineux sur l'image. Le procédé de détection d'un point lumineux sur une image consiste à : détecter un point lumineux sur une image à l'aide d'une matrice k1*k2, et déterminer qu'une matrice, dont une valeur de pixel central n'est pas inférieure à une première valeur prédéfinie, l'une quelconque des valeurs de pixel non central n'étant pas inférieure à une seconde valeur prédéfinie, et la valeur de pixel central n'étant pas inférieure à l'une quelconque des valeurs de pixel non central, correspond à un point lumineux candidat (S10), k1 et k2 étant tous deux des nombres impairs supérieurs à 1, et la matrice k1*k2 comprenant k1*k2 points de pixel ; et déterminer si le point lumineux candidat est un point lumineux (S20). Ledit procédé permet la détection rapide, efficace et précise d'un point lumineux sur une image, en particulier pour des images acquises à partir d'une réaction de mesure de séquence d'acides nucléiques.
PCT/CN2018/101817 2018-08-22 2018-08-22 Procédé et dispositif de détection d'un point lumineux sur une image, et procédé et dispositif d'enregistrement d'image WO2020037572A1 (fr)

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