A method and system for predicting organ weight of dwarf clams based on image grayscale value

Abstract

The present invention discloses a method and system for predicting the weight of dwarf clam organs based on image grayscale values. This method belongs to the field of organ measurement technology for small model organisms. The method comprises the following steps: capturing images of dwarf clam organs and extracting grayscale values; fitting a standard curve based on organ weights; and using the slope of the curve as a criterion for evaluating the growth stage of the dwarf clam organs; and predicting the weight of the dwarf clam organs by collecting the grayscale values ​​of the organs to be measured based on the standard curve. The method proposed in the present invention has the advantages of being simple, rapid, requiring no complex equipment, and being highly accurate. It can be widely applied to measuring the weight of organs in small organisms.

Description

Technical Field

[0001] The present invention relates to the technical field of small-scale model organism organ measurement, and in particular to a method and system for predicting the weight of dwarf clam organs based on image grayscale values. Background Art

[0002] Dwarf Clam ( Mulinia lateralis ) belongs to the family Cercoidea, order Bivalvia, phylum Mollusca. It is naturally distributed along the Atlantic coast of North America and the Caribbean, with a lifespan of approximately two years. Due to its rapid sexual maturity (2-3 months) and ease of laboratory propagation, the dwarf clam is considered an ideal model for shellfish genetic research. Our research group has established a standardized laboratory propagation system for dwarf clams, enabling five consecutive generations per year, providing material support for dwarf clam research.

[0003] Accurately measuring the weight of dwarf clams' organs is crucial for studying their growth, resistance, and reproduction. However, due to their small size, the weight of their organs is small and their shapes are irregular, making existing methods difficult to quickly and accurately measure. Currently, for large objects, volume is estimated by building three-dimensional models from captured images. For tobacco stems, which have regular shapes and minimal deformation, weight can be measured using pixel area. However, these methods are suitable for large organisms or objects with regular shapes and minimal deformation. The irregular shapes and uneven thickness of dwarf clam organs preclude similar methods. For small and medium-sized organisms, computer vision systems can measure the weight of individual crucian carp, but they cannot accurately measure organs. In fruit flies, organ size can be measured using microscopic imaging after specific antibodies are used to label organ regions. In New Zealand white rabbits, tissue density is estimated by obtaining grayscale images using a CT scanner. In Escherichia coli, the volume and dry weight of individual bacterial cells are measured using electron microscopy and integrated optical density image analysis. However, the above methods and equipment are not suitable for measuring the tiny organs of dwarf clams due to limitations such as high equipment requirements, cumbersome operations, complex analysis, and the need for specific antibodies.

[0004] Therefore, a method is needed that is simple to operate, does not require complicated equipment, measures accurately, is inexpensive, and can quickly measure the weight of dwarf clam organs. Summary of the Invention

[0005] In order to solve the above problems, the purpose of the present invention is to provide a method for measuring the weight of dwarf clam organs based on grayscale values, thereby solving the technical problem that the weight of small biological organs is difficult to measure.

[0006] In order to achieve the above technical objectives, the present application provides a method for predicting the weight of dwarf clam organs based on image grayscale values, comprising the following steps:

[0007] Images of dwarf clam organs were collected and grayscale values ​​were extracted. A standard curve was fitted based on the total weight and total grayscale value of each group of dwarf clam organs, and the slope of the curve was used as the standard for evaluating the growth stage of the dwarf clam organs.

[0008] Based on the standard curve, the weight of the dwarf clam organ to be tested is predicted by collecting the gray value of the dwarf clam organ to be tested.

[0009] Preferably, in the process of collecting images of the dwarf clam organs, the dwarf clam digestive gland, gonad, adductor muscle, gill filaments, mantle and foot are taken as the dwarf clam organs for image collection.

[0010] Preferably, in the process of collecting images of the dwarf clam organs, the images are collected by a stereo microscope and a light transmittance tester, using only white light.

[0011] Preferably, in the process of extracting the grayscale value, the software scale is calibrated according to the scale of the captured image, the organ image is converted to 8-bit format, and the image color is flipped; the organ outline is selected, and the total grayscale value of the area is measured.

[0012] Preferably, in the process of obtaining the grayscale value, the grayscale value of the background image is removed according to the grayscale value of the photographed dwarf clam organ image to obtain the corrected organ grayscale value, and the organ grayscale values ​​used in the standard curve fitting and weight prediction are all the corrected organ grayscale values.

[0013] Preferably, in the process of converting the organ image into 8-bit format, the software scale is calibrated according to the scale of the captured image, the scale is aligned using line segments, and the organ image is converted into 8-bit format by the average method.

[0014] Preferably, in the process of fitting the standard curve, the standard curve is represented as a linear fit through the origin:

[0015] y=bx,

[0016] Where x is the grayscale value of the organ image, y is the organ weight, and b is the slope. The slope b is different for dwarf clam organs at different growth stages.

[0017] The present invention discloses a dwarf clam organ weight prediction system based on image grayscale value, comprising:

[0018] A data acquisition module, used for acquiring images of dwarf clam organs and extracting grayscale values;

[0019] a data processing module for fitting a standard curve according to the extracted grayscale values ​​and the organ weight, and using the slope of the curve as a standard for evaluating the growth stage of the dwarf clam organ;

[0020] The prediction module is used to predict the weight of the dwarf clam organ to be tested by collecting the gray value of the dwarf clam organ to be tested based on the standard curve.

[0021] Preferably, the data acquisition module is further used to capture images of the digestive gland, gonad, adductor muscle, gill filaments, mantle and foot of the dwarf clam as organs of the dwarf clam.

[0022] Preferably, the data processing module is further used to calibrate the software scale according to the scale of the captured image, convert the organ image into 8-bit format, and flip the image color; select the organ outline and measure the total grayscale value of the area.

[0023] The present invention discloses the following technical effects:

[0024] 1) The present invention has high precision. Results show that the difference between the predicted and actual adductor muscle values ​​is only about 4%. The accuracy of a commonly used electronic balance is 0.1 mg, while the weight of the dwarf clam adductor muscle is around 1 mg, resulting in an error of 10% or more.

[0025] 2) This invention uses grayscale values ​​to represent the transmittance of dwarf clam organs. The principle behind this is that the intensity of a light beam passing through an object decreases exponentially with thickness and is directly related to the mass per unit area. That is, the smaller the organ weight per unit area, the greater the transmittance and the smaller the grayscale value. Existing methods measure weight using images of objects under the same lighting conditions, where the objects have regular shapes, minimal deformation, and are thick. These methods rely on reflection rather than transmission. Dwarf clam organs have irregular shapes and small, uneven thicknesses, so projection rather than reflection is the primary method used. Therefore, existing methods cannot accurately represent organ weight.

[0026] 3) The method is simple to use: it requires only using a stereomicroscope to capture images of dwarf clam organs and then calculating their grayscale values ​​using open-source software such as ImageJ. No radioactive instruments or specialized wavelengths of light are required, nor are specialized procedures or steps required to obtain image grayscale values.

[0027] 4) This invention does not affect the subsequent use of organ materials. After taking samples and taking pictures, the materials can be directly used in other experiments.

[0028] 5) The present invention uses a linear fit through the origin to determine the standard curve of the dwarf clam organs. This avoids the situation where the intercept of the standard curve of the organs is negative, some organs are too small, and the grayscale value is too small, thus avoiding the situation where the weight measurement result is negative. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] 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. Obviously, the drawings described below are only 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 work.

[0030] Figure 1 3. This is a linear relationship diagram of the weight and grayscale value of the dwarf clam digestive glands of the present invention, wherein the black dots represent the total weight and total grayscale value of the dwarf clam digestive glands of each group;

[0031] Figure 2 3 is a linear relationship diagram of the weight and grayscale value of the dwarf clam gonads of the present invention, wherein the black dots represent the total weight and total grayscale value of the dwarf clam gonads of each group;

[0032] Figure 3 3 is a linear relationship diagram of the weight and grayscale value of the adductor muscle of the dwarf clam of the present invention, wherein the black dots represent the total weight and total grayscale value of the adductor muscle of each group of dwarf clam;

[0033] Figure 4 It is a schematic flow chart of the method described in the present invention. DETAILED DESCRIPTION

[0034] In order to make the purpose, technical solutions and advantages of the embodiments of the present application clearer, the technical solutions in the embodiments of the present application will be clearly and completely described below in conjunction with the drawings in the embodiments of the present application. Obviously, the described embodiments are only a part of the embodiments of the present application, rather than all the embodiments. The components of the embodiments of the present application generally described and shown in the drawings here can be arranged and designed in various different configurations. Therefore, the following detailed description of the embodiments of the present application provided in the drawings is not intended to limit the scope of the application for protection, but merely represents the selected embodiments of the present application. Based on the embodiments of the present application, all other embodiments obtained by those skilled in the art without making creative work are within the scope of protection of this application.

[0035] like Figure 1-4 As shown, the present invention provides a method for predicting the weight of dwarf clam organs based on image grayscale values, comprising the following steps:

[0036] Images of dwarf clam organs were collected and grayscale values ​​were extracted. A standard curve was fitted based on the total weight and total grayscale value of each group of dwarf clam organs, and the slope of the curve was used as the standard for evaluating the growth stage of the dwarf clam organs.

[0037] Based on the standard curve, the weight of the dwarf clam organ to be tested is predicted by collecting the gray value of the dwarf clam organ to be tested.

[0038] Further preferably, in the method for predicting the weight of dwarf clam organs provided by the present invention, in the process of collecting images of dwarf clam organs, the dwarf clam digestive gland, gonad, adductor muscle, gill filaments, mantle and foot are used as dwarf clam organs for image collection.

[0039] Further preferably, in the method for predicting the weight of dwarf clam organs provided by the present invention, during the process of collecting images of the dwarf clam organs, the images are collected through a stereo microscope and a light transmittance tester, using only white light.

[0040] Further preferably, in the method for predicting the weight of dwarf clam organs provided by the present invention, in the process of extracting the grayscale value, the software scale is calibrated according to the scale of the captured image, the organ image is converted into 8-bit format, and the image color is flipped; the organ contour is selected, and the total grayscale value of the area is measured.

[0041] Further preferably, in the method for predicting the weight of dwarf clam organs provided by the present invention, in the process of obtaining the grayscale value, the grayscale value of the background image is removed according to the grayscale value of the photographed dwarf clam organ image to obtain the corrected organ grayscale value, and the organ grayscale values ​​used for fitting the standard curve and weight prediction are both the corrected organ grayscale values.

[0042] Further preferably, in the method for predicting the weight of dwarf clams provided by the present invention, in the process of converting the organ image into 8-bit format, the software scale is calibrated according to the scale of the captured image, the scale is aligned using line segments, and the organ image is converted into 8-bit format by the average method.

[0043] Further preferably, in the method for predicting organ weight of dwarf clams provided by the present invention, in the process of fitting the standard curve, the standard curve is represented by a linear fit through the origin:

[0044] y=bx,

[0045] Where x is the grayscale value of the organ image, y is the organ weight, and b is the slope. The slope b is different for dwarf clam organs at different growth stages.

[0046] The present invention discloses a dwarf clam organ weight prediction system based on image grayscale value, comprising:

[0047] A data acquisition module, used for acquiring images of dwarf clam organs and extracting grayscale values;

[0048] a data processing module for fitting a standard curve according to the extracted grayscale values ​​and the organ weight, and using the slope of the curve as a standard for evaluating the growth stage of the dwarf clam organ;

[0049] The prediction module is used to predict the weight of the dwarf clam organ to be tested by collecting the gray value of the dwarf clam organ to be tested based on the standard curve.

[0050] Further preferably, the data acquisition module of the dwarf clam organ weight prediction system provided by the present invention is also used to capture images of the dwarf clam digestive gland, gonad, adductor muscle, gill filaments, mantle and foot as dwarf clam organs.

[0051] Further preferably, the data processing module of the dwarf clam organ weight prediction system provided by the present invention is also used to calibrate the software scale according to the scale of the captured image, convert the organ image into 8-bit format, and flip the image color; select the organ contour and measure the total grayscale value of the area.

[0052] Example 1: The present invention provides a method for measuring the weight of dwarf clam organs based on grayscale values, thereby solving the technical problem that it is difficult to measure the weight of small organism organs.

[0053] The dwarf clams described in the present invention are dwarf clams cultured in this laboratory, and are about 45 days old. The organ images described are taken with a stereomicroscope; and the organ weights described are measured with an analytical balance.

[0054] In order to achieve the above object, the present invention adopts the following technical solutions:

[0055] 1) Prepare seven groups of 10, 20, 30, 40, 50, 60, and 70 well-grown dwarf clams, glass slides, and a stereo microscope.

[0056] 2) Place individual dwarf clam organs on glass slides and photograph them using a stereomicroscope. Also, take a background image without the dwarf clam organs. Use Image J to calculate the corrected total grayscale value of each organ group to represent the organ's light transmittance. The corrected organ grayscale value is the grayscale value of the organ image minus the grayscale value of the organ region in the background image.

[0057] 3) Measure the total weight of each group of dwarf clam organs.

[0058] 4) Perform a linear fit between the total grayscale value and total weight of the organ to obtain a standard curve.

[0059] Example 2: The grayscale value-based method for measuring the weight of dwarf clam organs proposed in the present invention specifically includes the following contents:

[0060] 1) Preparation

[0061] Select 280 well-growing dwarf clams and randomly divide them into seven groups, each containing 10, 20, 30, 40, 50, 60, and 70 clams. Prepare slides and a stereo microscope. Set the magnification of the stereo microscope to 0.68x, the software scale to 4×, and adjust the exposure parameters to avoid overexposure or underexposure. Do not change the exposure parameters during subsequent filming.

[0062] 2) Grayscale value measurement

[0063] The digestive gland, gonad, and adductor muscle of each group of dwarf clams were placed separately on glass slides, with the organs spread out flat and without excessive overlap, and centered in the light. Clear images of the organs were captured using a stereomicroscope, along with images of the background without the organs. Exposure parameters, such as exposure time and aperture, were kept constant. Images were acquired at a size of 2,048 × 1,536 pixels at 8,430 dpi and saved in TIF format. The grayscale values ​​of the captured dwarf clam organ images were calculated using Image J. The grayscale values ​​of the background images were then subtracted to obtain the corrected organ grayscale values. Finally, the total corrected grayscale values ​​of the dwarf clam organs for each group were calculated.

[0064] Image processing methods:

[0065] Calibrate the software scale according to the scale of the captured image. Align the scale bar with the line segments, select Analyze → Set Scale, enter the scale bar length (1,000 um), and click OK.

[0066] Convert the organ image to 8-bit format, using the average method. Select Image → Type → 8-bit. Then select Edit → Invert to invert the image colors.

[0067] ③ Select Analyze → Set Measurements and check the indicators to be measured (Area, Mean gray value, integrated density).

[0068] ④ Use the wand tool to select the organ outline and click Analyze→Measure to measure.

[0069] 3) Weight measurement

[0070] Each group of dwarf clam organs was placed in a cryovial, and the total weight of each group was recorded using an analytical balance.

[0071] 4) Draw a standard curve

[0072] SPSS software was used to perform linear fitting on the 7 groups of corrected organ total gray value and total weight data, and the fitting required selecting the origin.

[0073] The standard curve of digestive gland gray value and weight is:

[0074] Whep = 2.905×10-12Vhep-gray

[0075] Where Vhep-gray is the grayscale value of the digestive gland after correction, and Whep is the weight of the digestive gland (g). The results of significance analysis showed that the linear regression was extremely significant (p < 0.0001), with a correlation coefficient R² of 0.997 and a residual sum of squares of 4.97×10-6, indicating a good degree of linear fit ( Figure 1 ).

[0076] The standard curve of gonad gray value and weight is:

[0077] Wgon= 5.041×10-12Vgon-gray

[0078] Where Vgon-gray is the corrected gonad gray value, and Wgon is the gonad weight (g). The results of significance analysis showed that the linear regression was extremely significant (p < 0.0001), the correlation coefficient R² was 0.990, and the residual sum of squares was 8.72×10-6, indicating that the degree of linear fit was good ( Figure 2 ).

[0079] The standard curve of adductor muscle gray value and weight is:

[0080] Wmus = 4.129×10-12Vmus-gray

[0081] Where Vmus-gray is the grayscale value of the adductor muscle after correction, and Wmus is the weight of the adductor muscle (g). The results of the significance analysis showed that the linear regression was extremely significant (p < 0.0001), with a correlation coefficient R² of 0.996 and a residual sum of squares of 3.40×10-6, indicating a good degree of linear fit ( Figure 3 ).

[0082] 5) Actual Measurement: When measuring the weight of dwarf clam organs, take an image of the organ to be measured and a background image of a different organ. Use Image J to calculate the corrected grayscale value. Using the drawn standard curve, calculate the weight of the organ to be measured based on the corrected grayscale value.

[0083] On the basis of the embodiment, in order to improve the accuracy of grayscale value calculation of different organs, a grayscale value calculation method of a color image can be used, and the most appropriate method can be selected by comparing the accuracy of different methods.

[0084] The examples in this specification only introduce the weight measurement method of the digestive gland, gonads and adductor muscles of the dwarf clam. Since the weight measurement method of the gill filaments, mantle and feet corresponds to the method disclosed in the examples, it is not described. Please refer to the examples in this specification for details.

[0085] The grayscale-based method for measuring the weight of dwarf clam organs, provided by the present invention, overcomes the problems of large errors, complex operation, and high equipment costs associated with traditional methods. The method involves inputting the grayscale values ​​of dwarf clam organs into a standard curve of grayscale value and weight to obtain the weight of the organ to be measured. The standard curve is determined by capturing images of dwarf clam organs using a stereomicroscope, calculating the image grayscale values ​​using Image J, measuring the organ weights, and finally fitting a standard curve based on the total weight and total grayscale value of each organ group. This method is simple, rapid, requires no complex equipment, and offers high accuracy, making it widely applicable for measuring the weight of organs in small organisms.

[0086] The present invention is described with reference to flowcharts and / or block diagrams of methods, devices (systems), and computer program products according to embodiments of the present invention. It should be understood that each process and / or block in the flowcharts and / or block diagrams, as well as combinations of processes and / or blocks in the flowcharts and / or block diagrams, can be implemented by computer program instructions. These computer program instructions can be provided to a processor of a general-purpose computer, a special-purpose computer, an embedded processor, or other programmable data processing device to produce a machine, so that the instructions executed by the processor of the computer or other programmable data processing device generate instructions for implementing the processes in the flowcharts and / or block diagrams. Figure 1 a process or multiple processes and / or boxes Figure 1 A device that provides the functions specified in a block or multiple blocks.

[0087] In the description of the present invention, it should be understood that the terms "first" and "second" are used for descriptive purposes only and should not be understood to indicate or imply relative importance or implicitly specify the number of the technical features indicated. Therefore, a feature specified as "first" or "second" may explicitly or implicitly include one or more of the features. In the description of the present invention, "plurality" means two or more, unless otherwise specifically defined.

[0088] Obviously, those skilled in the art may make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if such changes and modifications fall within the scope of the claims and their equivalents, the present invention is intended to include such changes and modifications.

Claims

1. A dwarf clam organ weight prediction system based on image grayscale value, characterized by: It includes data acquisition module, data processing module and prediction module; The data acquisition module is used to collect images of dwarf clam organs and extract grayscale values ​​through a stereo microscope and a light transmittance tester, using only white light. The organs include a digestive gland, gonads, adductor muscles, gill filaments, mantle, and feet. The grayscale value extraction includes: calibrating the software scale according to the scale of the captured image; converting the organ image into 8-bit format and flipping the image color, wherein the 8-bit format conversion is achieved by aligning the scale with line segments and then using the average method; selecting the organ outline and measuring the total grayscale value of the area; The grayscale value is a corrected grayscale value, which is obtained by removing the grayscale value of the background image from the grayscale value of the captured dwarf clam organ image; The data processing module is used to fit a standard curve according to the extracted grayscale value and organ weight, and use the slope of the curve as a standard for evaluating the growth stage of the dwarf clam organ, wherein the standard curve is represented by a linear fitting curve passing through the origin, and its expression is y=bx, wherein x is the grayscale value of the captured organ image, y is the organ weight, and b is the slope. The slope b is different for dwarf clam organs at different growth stages; Among them, the standard curve of digestive gland gray value and weight is: Whep = 2.905×10 -12 Vhep-gray Where Vhep-gray is the gray value of the digestive gland after correction, and Whep is the weight of the digestive gland; The standard curve of gonad gray value and weight is: Wgon= 5.041×10 -12 Vgon-gray Where Vgon-gray is the corrected gonad gray value, and Wgon is the gonad weight; The standard curve of adductor muscle gray value and weight is: Wmus = 4.129×10 -12 Vmus-gray Where Vmus-gray is the corrected adductor muscle gray value, and Wmus is the adductor muscle weight; The prediction module is used to predict the weight of the dwarf clam organ to be tested by collecting the gray value of the dwarf clam organ to be tested based on the standard curve; When the prediction module performs weight prediction, the following steps are performed: capturing an image of the organ to be tested and background images of different organs; calculating the corrected grayscale value thereof using Image J; and calculating the weight of the organ to be tested according to the corrected grayscale value thereof using the drawn standard curve.

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