Heavy ion track positioning method suitable for tiny seeds

Abstract

The invention relates to the technical field of particle track microscopic analysis, in particular to a heavy ion track positioning method suitable for tiny seeds. The method mainly comprises the following steps of: scanning biological stacks by using a scanner, processing a scanning image by using drawing software to obtain an image of a single seed hole, acquiring position information of seeds in the seed holes, scanning a CR39 detection sheet by using a VHX microscope, acquiring passive detection information of each seed hole, and superposing the passive detection information with the image processed by the drawing software to obtain a final detection result. The radiation detection condition at the arabidopsis thaliana seeds is analyzed, and data conversion is further carried out to obtain various radiation information of individuals aiming at the arabidopsis thaliana seeds. The method is applicable to batch, rapid and high-precision processing of returned detection piece data. And in the aspect of the batch degree, the whole process is batch processing, and batch pictures and data can be automatically output by utilizing related software, so that the manual participation degree is reduced.

Description

Technical Field

[0001] The present invention relates to the technical field of particle track microanalysis, and in particular to a heavy ion track positioning method suitable for tiny seeds. Background Art

[0002] The space environment contains numerous particles, and solid-state nuclear track detectors are often used to analyze radiation parameters of these particles. The CR39 is a type of solid-state nuclear track detector designed to capture the tracks of radiation particles. After space flight, the detector sheet is recovered and etched, enabling the labeling and measurement of heavy ion tracks under an optical microscope. After calibration, software is used to automatically acquire heavy ion radiation nuclear track data. By analyzing the nuclear track parameters of the projected area of ​​Arabidopsis seeds on the CR39, radiation detection analysis can be performed on individual Arabidopsis seeds.

[0003] As interest in deep space exploration grows, the flight times and frequency of probes used in space missions are increasing. Earlier solutions, with their long durations and low accuracy, are no longer suitable for current practical applications. Earlier solutions also had significant shortcomings in analyzing heavy ions at the individual level of Arabidopsis seeds: the small size of Arabidopsis seeds made it impossible to accurately locate specific regions within the seeds. These three issues may impact future research. Summary of the Invention

[0004] In response to the aforementioned technical problems, a heavy ion track location method suitable for tiny seeds is provided. The present invention primarily uses a scanner to scan a biological stack, then processes the scanned image using mapping software to obtain an image of a single seed well and acquire the positional information of the seeds within the seed well. A VHX microscope is then used to scan a CR39 probe sheet, acquiring passive detection information for each seed well and overlaying it with the image processed by the mapping software. Radiation detection at the Arabidopsis seeds is analyzed, and further data conversion is performed to obtain various radiation information specific to each individual Arabidopsis seed.

[0005] The technical means adopted in the present invention are as follows: A heavy ion track positioning method suitable for tiny seeds, comprising: Obtain the projection data of the seed stack on the detection sheet and draw a pattern diagram of the seed hole position projection on the detection sheet; Under the microscope, focus twice on the same plane to obtain the passive microscopic detection image of the radiation corresponding to the detection piece at the transparent mask, and obtain the radiation detection information of the seed hole position in each model image; Obtain a physical image of the seeds through a scanner, and use a slicing tool to cut the physical image of the seeds into individual seed images; The object recognition function of the mapping software is used to identify the seed position in the single seed image to obtain the seed hole template image; The passive microscopic detection image of radiation corresponding to the detection piece at the transparent mask is superimposed on the seed template image to obtain the passive detection situation of radiation corresponding to the seed position; The number of heavy ion hits, radiation absorbed dose and LET value of each seed were calculated.

[0006] Furthermore, the acquisition of projection data of the seed stack on the detection sheet and the drawing of a pattern diagram of the seed hole positions projected on the detection sheet specifically include: The seeds are stacked in the seed holes, and the detection sheet is placed on the stack to obtain the projection of the stack on the detection sheet. A drawing is drawn based on the projection to obtain a frame of the same size as the detection sheet. The pattern diagram of the seed hole position projection on the detection sheet is drawn according to the stacking model data through image processing.

[0007] Furthermore, the microscope focuses twice on the same plane to obtain radiation passive microscopic detection images of the detection piece corresponding to the transparent mask, and obtains radiation detection information of the seed hole position in each model image, specifically including: Under a microscope, the method of focusing twice on the same plane was used to obtain a passive microscopic detection image of radiation from the detection piece corresponding to the transparent mask. The printed mask film and the detection piece were overlapped and pasted with 3M tape and then photographed under a microscope. The automatic movement distance of the stage was set according to the model image data using the electric stage and film parameters, and the radiation detection information in the seed holes in each model image was obtained by photographing.

[0008] Furthermore, the method of obtaining the seed physical image by a scanner and cutting the seed physical image into individual seed images by a slicing tool specifically includes: Use a scanner to scan the seed stack to obtain a clear seed physical image, add horizontal and vertical reference lines to the seed physical image so that a single hole can be located within a single reference line grid, use a slicing tool to slice based on the reference lines, and cut the entire seed image into individual seed images; the reference lines are drawn along the outer edges of the seed hole positions in the seed physical image to separate the seed hole positions from the intervals between the two holes, and the images are cut into small square pieces. The center of the square can coincide with the center of the circular hole to find the center of the circular hole.

[0009] Furthermore, the method of using the object recognition function to identify the seed position in the single seed image to obtain the seed hole template image specifically includes: The object recognition function is used to identify the positions of seeds inside the seed holes. After batch processing of the images, the seeds inside the seed holes are removed and the image background is converted to white to obtain template images of transparent seed holes at each seed position.

[0010] Furthermore, the passive microscopic detection image of radiation corresponding to the detection piece at the transparent mask is superimposed on the seed template image to obtain the passive detection situation of radiation corresponding to the seed position, specifically including: Focusing twice on the same plane, the radiation passive microscopic detection images corresponding to the detection piece at the transparent mask are superimposed with the template images of the corresponding transparent seed holes at each seed to obtain the radiation passive detection images corresponding to the seed positions.

[0011] Compared with the prior art, the present invention has the following advantages: The heavy ion track location method for tiny seeds, provided by this invention, is dedicated to batch processing of returned probe data, rapidly, and with high precision. The entire process is batch processed, and software can be used to automatically output batches of images and data, reducing manual intervention.

[0012] The heavy ion track location method for tiny seeds provided by this invention is highly efficient, allowing data collection time for a single CR-39 probe to be kept to just five man-hours. This method collects approximately 5,000 heavy ion tracks per probe. Compared to the biological stacking method used by the recoverable Shijian-10 satellite, which processed 10 CR-39 probes and collected fewer than 1,000 data points (individual heavy ion tracks), this method took 606 days, an average of approximately 60 days per probe, and involved three people (approximately 1,080 man-hours per probe, with each person using six man-hours per day). This method, compared to existing methods, increases the processing speed of a single probe by approximately 210 times and the number of track detections per man-hour by approximately 60 times.

[0013] The heavy ion track positioning method for tiny seeds provided by the present invention is accurate in that it uses mapping software to batch process and obtain the range of Arabidopsis seeds and obtain templates, which solves the problem that the range of Arabidopsis seeds cannot be accurately obtained in batches due to the small size of Arabidopsis seeds. The range of single Arabidopsis seeds can be obtained quickly, accurately and more simply.

[0014] Based on the above reasons, the present invention can be widely promoted in fields such as particle track microanalysis technology. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative labor.

[0016] Figure 1This is a flow chart of the heavy ion track positioning method applicable to tiny seeds in the present invention. DETAILED DESCRIPTION

[0017] It should be noted that, in the absence of conflict, the embodiments and features of the embodiments of the present invention can be combined with each other. The present invention will be described in detail below with reference to the accompanying drawings and in combination with the embodiments.

[0018] In order to make the purpose, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. The following description of at least one exemplary embodiment is actually only illustrative and is in no way intended to limit the present invention and its application or use. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of the present invention.

[0019] It should be noted that the terms used herein are only for describing specific embodiments and are not intended to limit the exemplary embodiments according to the present invention. As used herein, unless the context clearly indicates otherwise, the singular form is intended to include the plural form. In addition, it should be understood that when the terms "comprise" and / or "include" are used in this specification, they indicate the presence of features, steps, operations, devices, components and / or combinations thereof.

[0020] Unless otherwise specified, the relative arrangement of the parts and steps, numerical expressions and numerical values ​​set forth in these embodiments do not limit the scope of the present invention. At the same time, it should be clear that, for ease of description, the sizes of the various parts shown in the accompanying drawings are not drawn according to actual proportional relationships. The technology, methods and equipment known to those of ordinary skill in the relevant art may not be discussed in detail, but in appropriate cases, the technology, methods and equipment should be considered as a part of the specification. In all examples shown and discussed here, any specific value should be interpreted as being merely exemplary, rather than as a limitation. Therefore, other examples of the exemplary embodiments can have different values. It should be noted that similar numbers and letters represent similar items in the following drawings, and therefore, once an item is defined in an accompanying drawing, it does not need to be further discussed in subsequent drawings.

[0021] like Figure 1 As shown, the present invention provides a heavy ion track positioning method suitable for tiny seeds, characterized by comprising: Obtain projection data of the seed stack on the detector sheet and use SolidWorks to draw a diagram of the seed hole positions projected onto the detector sheet. In specific implementation, as a preferred embodiment of the present invention, the seeds are stacked in the seed holes and the detector sheet is placed on the stack to obtain a projection of the stack on the detector sheet. Based on the projection, a drawing is drawn to create a frame the same size as the detector sheet. Using image processing software (Adobe Illustrator), a diagram of the seed hole positions projected onto the detector sheet is drawn based on the stack model data and a transparent mask is printed. This step requires 0.5 man-hours and is performed only once; re-measurement is required when stacking or replacing the detector sheet.

[0022] Under a microscope, focus twice on the same plane to obtain passive microscopic radiation detection images of the probe sheet corresponding to the transparent mask, thereby obtaining radiation detection information for each seed hole within the model diagram. In a specific implementation, as a preferred embodiment of the present invention, the method of focusing twice on the same plane is used to obtain passive microscopic radiation detection images of the probe sheet corresponding to the transparent mask. The printed mask film and the probe sheet are then superimposed and affixed with 3M tape and photographed under a microscope. The motorized stage is used to automatically move the stage according to the model diagram data and the film parameters to obtain radiation detection information for each seed hole within the model diagram. This step requires four man-hours per sheet of 550 seeds.

[0023] Obtain the seed physical map by a scanner, and utilize a slicing tool to cut the seed physical map into a single seed picture; During specific implementation, as a preferred embodiment of the present invention, utilize a scanner to scan the seed stack, obtain a clear seed physical map, import software Adobe Photoshop to add horizontal and vertical reference lines to the seed physical map so that a single hole can be in a single reference line grid, utilize a slicing tool based on reference line slices, and cut the overall seed map into a single seed picture; Reference lines are drawn along the outer edge of the seed hole position in the seed physical map, separating the seed hole position from the interval of the two holes, and cutting into square small pieces of pictures, so that the square center can overlap with the circular hole center to find the circular hole center. This step requires 0.5 man-hours.

[0024] The object recognition function of Photoshop graphics software is used to identify the seed positions in a single seed image to obtain a seed hole template image. In a specific implementation, as a preferred embodiment of the present invention, the object recognition function is used to identify the positions of seeds within the seed holes. After batch processing of the images, the seeds within the seed holes are removed and the image background is converted to white to obtain template images of transparent seed holes at each seed location. This step is a batch automatic recognition process and does not require human time.

[0025] The passive microscopic radiation detection images corresponding to the transparent mask probe obtained twice by focusing on the same plane are superimposed with the seed template image to obtain the passive radiation detection situation corresponding to the seed position. In specific implementation, as a preferred embodiment of the present invention, the passive microscopic radiation detection images corresponding to the transparent mask probe obtained twice by focusing on the same plane are superimposed with the template image of the transparent seed hole at each seed position to obtain the passive radiation detection situation image corresponding to the seed position. This step is an automatic superposition process and is not performed manually.

[0026] A convolutional neural network model is used to capture particles in the overlay image, calculating the number of heavy ion hits, radiation absorbed dose, and particle LET value for each seed. This method is implemented using the "Two-Stage Automated Radiation Particle Track Acquisition Method and System for Solid-State Nuclear Track Detectors" (Application No. 2024105074274). The radiation particle dataset is preprocessed, manually marked major and minor axis points are corrected, and the heavy ion track data is normalized. The yolov7 model is used as a detector to extract image features and obtain the bounding rectangles of multiple radiation particles. An image of the roughly located rectangular frame is extracted, and ROIAglin is used to normalize rectangular frames of different sizes to the same representation space, amplifying the radiation particle features within the rectangular frame. A measuring device is used to extract characteristic information of the radiation particles and predict their tracks. The predicted relative center point, major and minor axis, and angles are restored to their absolute center point, major and minor axis, and angles relative to the image, achieving a heavy ion track localization method suitable for tiny seeds.

[0027] Example like Figure 1 As shown, in this embodiment, a scanner is used to scan the biological stack, and the scanned image is processed using mapping software to obtain an image of a single seed well and obtain the position information of the seeds in the seed well. A VHX microscope is used to scan the CR39 detection piece to obtain passive detection information of each seed well and overlay it with the image processed by the mapping software. The radiation detection situation at the Arabidopsis seeds is analyzed, and further data conversion is performed to obtain various radiation information for individual Arabidopsis seeds.

[0028] Arabidopsis seeds are small and numerous, and manual delineation of the seed range is time-consuming. Therefore, a program was developed using the batch processing and object selection functions in Adobe Photoshop software to batch process images to obtain templates. The resulting seed templates were then superimposed with subsequent radiation detection information from specific areas of the probe film to obtain radiation information for individual Arabidopsis seeds.

[0029] Heavy ions hitting the CR39 detector sheet leave tracks on the sheet. After etching, small holes can be observed on the surface of the sheet under a microscope. By calculating the area of ​​the small holes, various parameters of each heavy ion can be obtained. After the biological stack returns, the detector sheet is attached to a transparent mask that has been pre-drawn and printed based on the projection data of the biological stack on the detector sheet using 3M tape. Then, a VHX microscope is used to focus twice on the same plane to obtain a passive microscopic detection image of the radiation from the detector sheet corresponding to the transparent mask, and to photograph the passive detection situation at each seed hole position. Focusing twice on the same plane to obtain a passive microscopic detection image of the radiation from the detector sheet corresponding to the transparent mask means first focusing on the mask surface, locating the seed hole according to the seed hole information marked on the mask, and then refocusing on the CR39 detector sheet without changing the position to achieve the purpose of photographing the detection situation at the seed hole position.

[0030] The microscope used in this method is a VHX microscope, which is equipped with a motorized stage adapted to the microscope. The distance of each lateral or longitudinal movement can be set by the user to achieve semi-automatic shooting. The distances of lateral and longitudinal movement are pre-set during shooting, and the distance parameters are consistent with the projection data of the organism superimposed on the detection piece. The microscope shoots the side where the detection piece and the seed are attached, with a shooting magnification of 200 times, and the focal length is adjusted to make the field of view clear. Adjust the light intensity to avoid the field of view being too bright or too dark. After superimposing the acquired image and the template one by one, use the automated scanning and reading software customized for this data format to read and obtain the measurement data of the number of heavy ion tracks and the major and minor axes.

[0031] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the above embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the above embodiments, or replace some or all of the technical features therein with equivalents. However, these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A heavy ion track positioning method suitable for tiny seeds, characterized in that: include: Obtain the projection data of the seed stack on the detection sheet and draw a pattern diagram of the seed hole position projection on the detection sheet; Under the microscope, focus twice on the same plane to obtain the passive microscopic detection image of the radiation corresponding to the detection piece at the transparent mask, and obtain the radiation detection information of the seed hole position in each model image; Obtain a physical image of the seeds through a scanner, and use a slicing tool to cut the physical image of the seeds into individual seed images; The object recognition function of the mapping software is used to identify the seed position in the single seed image to obtain the seed hole template image; The passive microscopic detection image of radiation corresponding to the detection piece at the transparent mask is superimposed on the seed template image to obtain the passive detection situation of radiation corresponding to the seed position; The number of heavy ion hits, radiation absorbed dose and LET value of each seed were calculated.

2. The heavy ion track positioning method for tiny seeds according to claim 1, characterized in that: The step of obtaining projection data of the seed stack on the detection sheet and drawing a pattern diagram of the seed hole positions projected on the detection sheet specifically includes: The seeds are stacked in the seed holes, and the detection sheet is placed on the stack to obtain the projection of the stack on the detection sheet. A drawing is drawn based on the projection to obtain a frame of the same size as the detection sheet. The pattern diagram of the seed hole position projection on the detection sheet is drawn according to the stacking model data through image processing.

3. The heavy ion track positioning method for tiny seeds according to claim 1, characterized in that: The microscope focuses on the same plane twice to obtain the passive microscopic detection images of radiation corresponding to the detection piece at the transparent mask, and obtains the radiation detection information of the seed hole position in each model image, specifically including: Under a microscope, the method of focusing twice on the same plane was used to obtain a passive microscopic detection image of radiation from the detection piece corresponding to the transparent mask. The printed mask film and the detection piece were overlapped and pasted with 3M tape and then photographed under a microscope. The automatic movement distance of the stage was set according to the model image data using the electric stage and film parameters, and the radiation detection information in the seed holes in each model image was obtained by photographing.

4. The heavy ion track positioning method for tiny seeds according to claim 1, characterized in that: The method of obtaining a seed physical image by a scanner and cutting the seed physical image into individual seed images by a slicing tool specifically includes: Use a scanner to scan the seed stack to obtain a clear seed physical image, add horizontal and vertical reference lines to the seed physical image so that a single hole can be located within a single reference line grid, use a slicing tool to slice based on the reference lines, and cut the entire seed image into individual seed images; the reference lines are drawn along the outer edges of the seed hole positions in the seed physical image to separate the seed hole positions from the intervals between the two holes, and the images are cut into small square pieces. The center of the square can coincide with the center of the circular hole to find the center of the circular hole.

5. The heavy ion track positioning method for tiny seeds according to claim 1, characterized in that: The method of using the object recognition function to identify the seed position in a single seed image to obtain a seed hole template image specifically includes: The object recognition function is used to identify the positions of seeds inside the seed holes. After batch processing of the images, the seeds inside the seed holes are removed and the image background is converted to white to obtain template images of transparent seed holes at each seed position.

6. The heavy ion track positioning method for tiny seeds according to claim 1, characterized in that: The passive microscopic detection image of radiation corresponding to the transparent mask detection piece is superimposed on the seed template image to obtain the passive detection situation of radiation corresponding to the seed position, specifically including: Focusing twice on the same plane, the radiation passive microscopic detection images corresponding to the detection piece at the transparent mask are superimposed with the template images of the corresponding transparent seed holes at each seed to obtain the radiation passive detection images corresponding to the seed positions.

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