Method for identifying rice processing precision

Abstract

The application discloses a method for identifying rice processing precision, and relates to the technical field of rice processing, which comprises the following steps: determining the groove area of a rice kernel according to the back-groove bright-field image of the rice kernel; generating a three-section reagent sequence application record according to the three-section reagent sequence and the groove area of the rice kernel; obtaining a liquid front advance sequence according to the three-section reagent sequence application record; calculating the channel focusing degree according to the central narrowband area and the sideband area of the fluorescence image sequence; and calculating the membrane retention focusing value according to the first average fluorescence intensity, the second average fluorescence intensity and the channel focusing degree of the groove fluorescence intensity sequence. The application can improve the pertinence and objectivity of rice processing precision detection.

Description

Technical Field

[0001] This invention relates to the field of rice processing technology, and in particular to a method for determining the precision of rice processing. Background Technology

[0002] During rice processing, as milling continues, the bran layer, aleurone layer, and related residual tissues adhering to the surface of the rice grains are gradually removed. However, the dorsal groove region of the rice grain, due to its longitudinal concave structure, narrow space, and complex surface morphology, often becomes the area where residual tissues are most prominently retained. The distribution, continuity, and retention of residual tissues in the dorsal groove region can directly reflect the actual processing status of the rice. In the current rice processing quality evaluation process, it is necessary to conduct processing precision testing on a large number of rice samples to meet the application needs of grain processing enterprises for quality control, grain storage and inspection, market circulation grading, and scientific research analysis.

[0003] In existing technologies, the determination of rice processing precision mainly relies on manual observation of residual tissue on the surface of rice grains, visual comparison after dyeing, or evaluation of the processing status through overall appearance indicators. Such methods are difficult to accurately reflect the true distribution of residual tissue inside the back groove area. Since the back groove area is the part where residual tissue is most easily retained during processing, it is difficult to quantify the retention capacity, spatial continuity, and liquid entry into the back groove channel based solely on appearance observation results. Therefore, it is easily affected by factors such as differences in the experience of observers, individual differences in samples, and uneven distribution of local residual tissue, resulting in insufficient stability of the test results and making it difficult to accurately identify the processing precision of rice. Summary of the Invention

[0004] The purpose of this invention is to address the shortcomings of existing technologies in accurately assessing the precision of rice processing, and to propose a method for assessing the precision of rice processing.

[0005] To address the problems existing in the prior art, the present invention adopts the following technical solution:

[0006] Methods for determining the precision of rice processing include:

[0007] S1. Determine the dorsal groove region of the rice grain based on the bright-field image of the dorsal groove of the rice grain.

[0008] S2. Based on the three-segment reagent sequence and the dorsal groove region of the rice grain, generate a three-segment reagent sequence and apply it for recording;

[0009] S3. Based on the three-segment reagent sequence, a recording is applied to obtain the liquid front advancement sequence;

[0010] S4. Calculate the channel focus based on the central narrow band region and the side band region of the fluorescence image sequence; calculate the membrane retention focus value based on the first average fluorescence intensity, the second average fluorescence intensity, and the channel focus based on the back groove fluorescence intensity sequence.

[0011] S5. Based on the membrane retention focusing value and the liquid front propulsion sequence, the accuracy of the rice sample to be tested is judged, and the judgment result is obtained.

[0012] Preferably, determining the dorsal groove region of the rice grain includes:

[0013] Obtain the set of rice grains to be tested from the rice sample of the batch to be tested;

[0014] Acquire a bright-field image of the back groove of each rice grain in the set of rice grains to be tested;

[0015] In the bright-field image of the back gutter, determine the inlet point, end point, and centerline of the back gutter;

[0016] Determine the boundary of the back groove in the bright-field image of the back groove;

[0017] The back groove region of the rice grain is determined based on the inlet point, end point, center line, and boundary of the back groove.

[0018] Preferably, generating a three-segment reagent sequence and applying recording includes:

[0019] A three-stage reagent sequence was configured; wherein the three-stage reagent sequence includes an aqueous wetting solution, a lipid affinity probe solution, and a mild replacement solution;

[0020] Apply the aqueous wetting solution to the inlet point of the dorsal groove of each rice grain, so that the aqueous wetting solution enters the dorsal groove area of ​​the rice grain along the center line of the dorsal groove.

[0021] After the aqueous wetting solution is applied, the lipid affinity probe solution is applied to the dorsal groove inlet point of the same rice grain, so that the lipid affinity probe solution enters the dorsal groove region of the rice grain along the dorsal groove centerline.

[0022] After the lipid affinity probe solution is applied, the mild replacement solution is applied to the dorsal groove inlet point of the same rice grain, so that the mild replacement solution enters the dorsal groove region along the dorsal groove centerline;

[0023] Record the application time intervals of the aqueous wetting solution, lipid affinity probe solution, and mild replacement solution for the rice grains;

[0024] Based on the application time intervals of the aqueous wetting solution, the lipid affinity probe solution, and the mild replacement solution, a three-segment reagent sequence application record is generated.

[0025] Preferably, the liquid front propulsion sequence includes:

[0026] Within the time intervals of the aqueous wetting solution application, lipid affinity probe solution application, and mild replacement solution application recorded in the three-segment reagent sequence application, continuous bright-field image acquisition was performed on the dorsal groove region to obtain a bright-field image sequence.

[0027] Based on the centerline of the back groove, the position of the liquid front in the bright field image sequence is extracted to obtain the liquid front advancement sequence.

[0028] Preferably, calculating the channel focus includes:

[0029] During the time intervals of lipid affinity probe solution application and mild replacement solution application, continuous fluorescence images of the dorsal groove region were acquired. The fluorescence images obtained from each acquisition were arranged in chronological order of acquisition time to obtain a fluorescence image sequence.

[0030] Based on the dorsal groove centerline, the central narrow band region and lateral band region are determined in the fluorescence image sequence;

[0031] Based on the application time interval of the mild replacement fluid, the average fluorescence intensity of the central narrow band region was extracted from the fluorescence image sequence;

[0032] Based on the application time interval of the mild replacement fluid, the average fluorescence intensity of the sideband region was extracted from the fluorescence image sequence;

[0033] The channel focusing power is obtained by dividing the average fluorescence intensity of the central narrow band region by the average fluorescence intensity of the side band region.

[0034] Preferably, calculating the film retention focus value includes:

[0035] The fluorescence intensity at the corresponding position of the dorsal groove centerline is extracted frame by frame in the fluorescence image sequence to obtain the dorsal groove fluorescence intensity sequence.

[0036] Based on the application time interval of the lipid affinity probe solution, the first average fluorescence intensity was extracted from the dorsal groove fluorescence intensity sequence;

[0037] Based on the time interval of mild replacement fluid application, the second average fluorescence intensity was extracted from the dorsal groove fluorescence intensity sequence;

[0038] Divide the second average fluorescence intensity by the first average fluorescence intensity to obtain the retention ratio;

[0039] Multiply the retention ratio by the channel focusing degree to obtain the membrane retention focusing value.

[0040] Preferably, the judgment result includes:

[0041] Based on the application time interval of the lipid affinity probe solution, the maximum liquid front advancement length was extracted from the liquid front advancement sequence;

[0042] The propulsion occupancy ratio is calculated based on the maximum liquid front propulsion length and the length of the back groove centerline;

[0043] The processing accuracy assessment value is calculated based on the membrane retention focus value and the propulsion occupancy ratio.

[0044] Based on the processing precision identification value, the precision of the rice sample batch to be tested is judged, and the judgment result is obtained.

[0045] Preferably, determining the inlet point, end point, and centerline of the back gutter in the bright-field image includes:

[0046] Extract the outer contour of rice grains from the bright-field image of the back ditch;

[0047] Based on the outer contour of the rice grain, extract the dark groove area on the back;

[0048] Connectivity screening is performed on the dark texture region of the back groove to obtain candidate regions for the back groove;

[0049] The candidate regions of the back groove are skeletonized to obtain the candidate skeleton lines of the back groove.

[0050] One end of the candidate skeleton line of the back groove is determined as the entry point of the back groove, and the other end of the candidate skeleton line of the back groove is determined as the end point of the back groove.

[0051] The candidate skeleton line of the backditch connecting the inlet point and the end point of the backditch is determined as the center line of the backditch.

[0052] Preferably, determining the back groove boundary in the back groove bright-field image includes:

[0053] Along the direction perpendicular to the center line of the back groove, the dark texture area of ​​the back groove corresponding to the bright field image of the back groove is scanned point by point to obtain multiple horizontal grayscale distributions.

[0054] In each horizontal grayscale distribution, determine the grayscale change positions on both sides of the back groove center line;

[0055] Connecting multiple grayscale change locations on one side of the back groove centerline yields the first back groove boundary;

[0056] Connecting multiple grayscale change locations on the other side of the back groove centerline yields the second back groove boundary;

[0057] The first back groove boundary and the second back groove boundary are defined as back groove boundaries.

[0058] Compared with the prior art, the beneficial effects of the present invention are:

[0059] 1. This invention constructs a back groove region identification mechanism to determine the back groove inlet point, back groove end point, back groove center line and back groove boundary of rice grain, forming a corresponding back groove region. This shifts the detection target from the overall appearance of the rice grain to the concentrated distribution area of ​​back groove residual tissue, avoiding the judgment bias caused by relying solely on surface observation, and improving the pertinence and objectivity of rice processing precision detection.

[0060] 2. This invention employs a three-stage reagent sequence consisting of an aqueous wetting solution, a lipid affinity probe solution, and a mild replacement solution to act on the dorsal groove region. It combines the liquid front advancement process and fluorescence distribution process for analysis, which can simultaneously obtain the retention state of the probe substance by the residual tissue and the entry state of the liquid in the dorsal groove channel, thereby improving the characterization ability of the actual existence of residual tissue in the dorsal groove.

[0061] 3. This invention establishes a quantitative correlation between the state of residual tissue in the dorsal groove and processing precision by constructing channel focusing degree, retention ratio, membrane retention focusing value and processing precision identification value. This allows the test results to simultaneously reflect the degree of residual tissue retention, spatial distribution and dorsal groove channel openness, reducing the impact of individual sample differences on the test results and improving the stability and accuracy of rice processing precision identification results. Attached Figure Description

[0062] The accompanying drawings, which are included to provide a further understanding of the invention and form part of this application, illustrate exemplary embodiments of the invention and, together with their description, serve to explain the invention and do not constitute an undue limitation thereof. In the drawings:

[0063] Figure 1 This is a flowchart illustrating a method for determining the processing precision of rice according to an embodiment of the present invention. Detailed Implementation

[0064] The technical solutions of the present invention will be clearly and completely described below with reference to the accompanying drawings of the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments.

[0065] This embodiment provides a method for determining the precision of rice processing; see [link to documentation]. Figure 1 Specifically, including:

[0066] S1. Determine the dorsal groove region of the rice grain based on the bright-field image of the dorsal groove of the rice grain.

[0067] In an embodiment of the present invention, determining the dorsal groove region of a rice grain includes:

[0068] Obtain the set of rice grains to be tested from the rice sample of the batch to be tested;

[0069] The rice grain set to be tested refers to a sample set formed by multiple independent rice grains extracted from the same batch of rice samples to be tested. Each rice grain is used as an independent detection object to participate in subsequent analysis. The rice grain set to be tested is used to reflect the overall processing status and back groove residue status of the batch of rice samples to be tested.

[0070] The rice sample to be tested was placed on a clean support surface and dispersed to create gaps between adjacent grains that allowed for individual picking and observation. Foreign matter and non-rice particles were removed from the sample, and the remaining grains were retained regardless of whether they were broken or significantly damaged. Based on their actual shape, the remaining grains were categorized into intact grains, broken grains, and significantly damaged grains. Intact grains were defined as those with a continuous main outline and no obvious breakage; broken grains were those separated due to processing or other factors; and significantly damaged grains were those whose main body remained connected but had missing material in localized areas. The number of intact, broken, and significantly damaged grains was then counted. The actual number of remaining rice grains is determined, and multiple rice grains of each type are extracted according to their actual quantity in the remaining rice grains. This ensures that the extracted rice grains include intact grains, broken grains, and obviously damaged grains, and that the composition of each type of grain in the extracted results corresponds to the actual composition of the rice sample in the batch to be tested. Each intact grain, broken grain, and obviously damaged grain is treated as an independent testing object, and a corresponding grain label is established for each. Grains that are broken or partially damaged are not excluded from subsequent testing. All independent testing objects are then combined to form a set of rice grains to be tested, ensuring that the set includes rice grains of different shapes and states from the rice sample in the batch to be tested, thus preserving the rice grain composition formed during the actual processing of the rice sample in the batch to be tested.

[0071] Acquire a bright-field image of the back groove of each rice grain in the set of rice grains to be tested;

[0072] Specifically, each grain of rice in the set of rice grains to be tested is placed on a support fixture with a positioning groove. The posture of the rice grain is adjusted so that the back groove of the rice grain faces the imaging direction of the image acquisition device. The support fixture is adjusted so that the back groove of the rice grain is within the field of view of the image acquisition device. Visible light is applied to the back groove area of ​​the rice grain. After the image acquisition device completes focusing, a bright field image including the back groove area of ​​the rice grain, the two edge areas on both sides of the rice grain, and the two ends of the length direction of the rice grain is acquired. The acquired image is then correlated with the corresponding rice grain to obtain the back groove bright field image of each rice grain in the set of rice grains to be tested.

[0073] In the bright-field image of the back gutter, determine the inlet point, end point, and centerline of the back gutter;

[0074] Bright-field images of the dorsal groove refer to two-dimensional image data obtained by imaging the dorsal groove region of a rice grain using visible light illumination. These images record the brightness differences, grayscale distribution differences, and spatial relationships between the dorsal groove region and the surrounding endosperm region. The dorsal groove region typically appears as a concave band extending along the length of the rice grain. Bright-field images of the dorsal groove region are used to characterize its morphology and distribution. The dorsal groove inlet point refers to the boundary position of one end of the dorsal groove along its length. This position is the intersection of the dorsal groove centerline and the outline of one end of the rice grain. The dorsal groove inlet point corresponds to the point where subsequent liquid first contacts the dorsal groove region. The location of the ditch region is used to determine the starting point of the ditch region and the direction of its extension. The end point of the ditch refers to the other end boundary of the ditch along its length. This location is the intersection of the center line of the ditch and the outline of the other end of the rice grain. The end point of the ditch corresponds to the end point of the ditch region along its length. It is used to define the end point of the ditch region and the actual extension range of the ditch. The center line of the ditch refers to the reference line located at the center of the ditch region and continuously distributed along the direction of the ditch's extension. The center line of the ditch is formed by connecting the center positions of each cross-section of the ditch region in sequence. It reflects the main extension trajectory and spatial distribution direction of the ditch region.

[0075] In an embodiment of the present invention, determining the inlet point, end point, and centerline of the back gutter in a bright-field image includes:

[0076] Extract the outer contour of rice grains from the bright-field image of the back ditch;

[0077] Brightness equalization is performed on the bright field image of the back groove to maintain a distinguishable brightness difference between the background area and the grain area. The bright field image of the back groove after brightness equalization is then converted to grayscale to obtain the grayscale image of the back groove. The grain area and the background area are separated based on the difference in grayscale distribution between the background area and the grain area in the grayscale image of the back groove. The closed area formed by continuous grain pixels is retained, and the background pixels separated from the closed area and isolated noise pixels are removed. The boundary line is extracted along the outermost pixel edge of the closed area to obtain the outer contour of the grain.

[0078] Based on the outer contour of the rice grain, extract the dark groove area on the back;

[0079] The outer contour of the rice grain refers to the closed boundary formed by the contact between the rice grain and the background in the bright field image of the back groove. This closed boundary is used to define the actual area occupied by the rice grain in the image and to reflect the overall shape and spatial distribution range of the rice grain. The dark texture area of ​​the back groove refers to the low brightness area located inside the outer contour of the rice grain and extending continuously along the length of the rice grain. This area corresponds to the gray-scale reduction area formed by the back groove concave structure under bright field imaging conditions and is used to characterize the actual distribution position and extension state of the back groove on the surface of the rice grain.

[0080] The image area enclosed by the outer contour of the rice grain is defined as the inner region of the rice grain. Within the inner region of the rice grain, the grayscale changes at each position along the length of the rice grain are statistically analyzed. Pixel regions with grayscale values ​​lower than the adjacent endosperm region and extending along the length of the rice grain are retained. Dark areas located outside the outer contour of the rice grain, shadow areas located at the edge of the rice grain, and local dark spot areas that do not extend along the length of the rice grain are removed. The retained low-brightness extended areas are designated as the back groove dark texture areas.

[0081] Connectivity screening is performed on the dark texture region of the back groove to obtain candidate regions for the back groove;

[0082] Based on the contact relationship between adjacent pixels in the dark groove region, multiple separate dark groove connected regions are divided. For each dark groove connected region, its extension direction, region length, region width, and correspondence with the length direction of the rice grain are determined. Dark groove connected regions that extend along the length direction of the rice grain and are located in the middle of the internal region of the rice grain are retained. Dark groove connected regions that are distributed at the edge of the rice grain, are isolated points, are short horizontal patches, or do not correspond to the length direction of the rice grain are removed. The retained dark groove connected regions are determined as candidate regions for the back groove.

[0083] The candidate regions of the back groove are skeletonized to obtain the candidate skeleton lines of the back groove.

[0084] The candidate back groove region refers to the connected region selected from the dark back groove region. This region maintains the spatial morphology of the dark back groove region continuously distributed along the length of the grain and is used to represent the range of regions that may correspond to the back groove structure. The candidate back groove skeleton line refers to the central path line retained after shrinking the candidate back groove region. This path line is located inside the candidate back groove region and maintains the overall extension direction and topological structure of the candidate back groove region.

[0085] Specifically, the candidate back groove region is converted into a binary region composed of pixels of the candidate back groove region. The edge pixels of the candidate back groove region are shrunk layer by layer, so that the two sides of the candidate back groove region shrink towards the center of the region. During the shrinkage process, the connectivity and extension direction of the candidate back groove region along the length of the grain are preserved. The process stops when the candidate back groove region can no longer shrink but still maintains a continuous path. The continuous path located in the center of the candidate back groove region is determined as the candidate back groove skeleton line.

[0086] One end of the candidate skeleton line of the back groove is determined as the entry point of the back groove, and the other end of the candidate skeleton line of the back groove is determined as the end point of the back groove.

[0087] Specifically, the two endpoints of the candidate skeleton line for the back groove are determined, the image coordinate system in the bright field image of the back groove is determined, one side along the length of the grain in the image coordinate system is taken as the inlet side of the back groove, and the other side along the length of the grain in the image coordinate system is taken as the end side of the back groove. The endpoint of the candidate skeleton line located on the inlet side of the back groove is determined as the inlet point of the back groove, and the endpoint of the candidate skeleton line located on the end side of the back groove is determined as the end point of the back groove.

[0088] The candidate skeleton line of the backditch connecting the inlet point and the end point of the backditch is determined as the center line of the backditch.

[0089] Specifically, on the candidate skeleton line of the back groove, the continuous skeleton path between the inlet point and the end point of the back groove is retained, and the branched paths and isolated paths that do not connect the inlet point and the end point of the back groove are removed. The retained continuous skeleton path is used as the center line of the back groove, and the arrangement order of each position on the center line of the back groove is recorded so that the center line of the back groove can be used for subsequent extraction of the liquid front position and fluorescence intensity along the back groove direction.

[0090] Determine the boundary of the back groove in the bright-field image of the back groove;

[0091] In an embodiment of the present invention, determining the back gutter boundary in a bright-field image includes:

[0092] Along the direction perpendicular to the center line of the back groove, the dark texture area of ​​the back groove corresponding to the bright field image of the back groove is scanned point by point to obtain multiple horizontal grayscale distributions.

[0093] Specifically, continuous path points on the center line of the back groove are used as reference points for lateral scanning. At each reference point, the tangential direction of the back groove center line at that reference point is determined. The direction that intersects the tangential direction of the back groove center line and crosses the dark texture area of ​​the back groove is taken as the lateral scanning direction. The lateral scanning direction extends from one side of the back groove center line through the back groove center line to the other side. The pixel gray values ​​in the lateral scanning direction are obtained sequentially in the back groove bright field image. The multiple pixel gray values ​​obtained corresponding to the same reference point are arranged in the spatial order of the lateral scanning direction to form the lateral gray distribution corresponding to that reference point. The lateral gray distributions corresponding to each reference point are summarized into multiple lateral gray distributions.

[0094] In each horizontal grayscale distribution, determine the grayscale change positions on both sides of the back groove center line;

[0095] The horizontal grayscale distribution refers to the grayscale arrangement result obtained in a horizontal scan. This result reflects the brightness change of the back groove dark texture area and its adjacent areas on both sides in the horizontal direction. The grayscale change position refers to the position in the horizontal grayscale distribution where the brightness transitions from the darker state corresponding to the back groove depression to the brighter state corresponding to the rice grain surface on both sides. This position is used to indicate the edge position between the back groove depression area and the adjacent rice grain surface area.

[0096] Specifically, taking the position of the center line of the back groove in the horizontal gray-scale distribution as the center position, the gray-scale transitions continuously from the low gray-scale state corresponding to the dark texture area of ​​the back groove to the higher gray-scale state corresponding to the surface area of ​​the rice grain from the center position to both sides of the horizontal gray-scale distribution. The position closest to the center position on one side and located in the continuous transition section is determined as the gray-scale change position on that side. The position closest to the center position on the other side and located in the continuous transition section is determined as the gray-scale change position on the other side. The two gray-scale change positions obtained from the same horizontal gray-scale distribution are taken as the boundary positions of the back groove on both sides corresponding to the reference point.

[0097] Connecting multiple grayscale change locations on one side of the back groove centerline yields the first back groove boundary;

[0098] Specifically, according to the arrangement order of each reference point on the center line of the back groove from the entrance point to the end point of the back groove, the gray-scale change positions on the same side are arranged sequentially, and the gray-scale change positions on the same side corresponding to adjacent reference points are connected in sequence to form a boundary path that extends continuously along the length of the back groove. The isolated deviation positions caused by local image noise in the boundary path are corrected according to the extension direction of the adjacent gray-scale change positions, so that the corrected boundary path is kept at the gray-scale transition position between the dark texture area of ​​the back groove and the surface area of ​​the rice grain. The corrected boundary path is determined as the first back groove boundary.

[0099] Connecting multiple grayscale change locations on the other side of the back groove centerline yields the second back groove boundary;

[0100] Specifically, according to the arrangement order of each reference point on the center line of the back groove from the entrance point to the end point of the back groove, the gray-scale change positions on the other side are arranged sequentially. The gray-scale change positions on the other side corresponding to adjacent reference points are connected in sequence to form another boundary path that extends continuously along the length of the back groove. The isolated deviation positions caused by local image noise in the other boundary path are corrected according to the extension direction of the adjacent gray-scale change positions, so that the corrected other boundary path is kept at the gray-scale transition position between the dark texture area of ​​the back groove and the surface area of ​​the rice grain. The corrected other boundary path is determined as the second back groove boundary.

[0101] The first back groove boundary and the second back groove boundary are defined as back groove boundaries.

[0102] The first back groove boundary refers to the boundary line formed by connecting multiple gray-scale change positions located on one side of the back groove centerline in the direction of the extension of the back groove centerline; the second back groove boundary refers to the boundary line formed by connecting multiple gray-scale change positions located on the other side of the back groove centerline in the direction of the extension of the back groove centerline.

[0103] Specifically, the first back groove boundary is taken as the boundary line on one side of the back groove center line, and the second back groove boundary is taken as the boundary line on the other side of the back groove center line. The image area enclosed by the first back groove boundary, the second back groove boundary, the end position where the back groove entrance point is located, and the end position where the back groove end point is located is determined as the boundary range of the back groove region of the rice grain, and the first back groove boundary and the second back groove boundary are output as the back groove boundary of the rice grain.

[0104] The back groove region of the rice grain is determined based on the inlet point, end point, center line, and boundary of the back groove.

[0105] Specifically, the center line of the back groove is used as the length direction reference of the back groove region. The location of the back groove entrance point is determined as the starting end of the back groove region, and the location of the back groove end point is determined as the ending end of the back groove region. The first back groove boundary is used as the region boundary on one side of the back groove center line, and the second back groove boundary is used as the region boundary on the other side of the back groove center line. Along the direction from the back groove entrance point to the back groove end point, the first back groove boundary, the second back groove boundary, the end boundary connecting the first back groove boundary and the second back groove boundary at the back groove entrance point, and the end boundary connecting the first back groove boundary and the second back groove boundary at the back groove end point are used to enclose a closed image region. The pixel region within this closed image region is determined as the back groove region of the rice grain.

[0106] S2. Based on the three-segment reagent sequence and the dorsal groove region of the rice grain, generate a three-segment reagent sequence and apply it for recording;

[0107] In an embodiment of the present invention, generating a three-segment reagent sequence and applying recording includes:

[0108] A three-stage reagent sequence was configured; wherein the three-stage reagent sequence includes an aqueous wetting solution, a lipid affinity probe solution, and a mild replacement solution;

[0109] The three-stage reagent sequence refers to a combination of multiple reagents applied to the dorsal groove region in a fixed order, where each reagent acts sequentially on the same dorsal groove region, forming a continuous liquid interaction process; the aqueous wetting solution refers to a liquid medium formed with water as the continuous phase, which can enter the dorsal groove region and contact the surface of the dorsal groove region, forming a continuous liquid phase pathway for subsequent liquids to enter the dorsal groove region; the lipid affinity probe solution refers to a liquid medium containing probe substances that can interact with lipid components, which, after entering the dorsal groove region, can contact the lipid components corresponding to the residual tissue in the dorsal groove region and form a response signal; the mild replacement solution refers to a liquid medium that can replace the existing liquid in the dorsal groove region without damaging the overall structure of the dorsal groove region, which is used to change the liquid composition in the dorsal groove region while preserving the original spatial distribution of the dorsal groove region;

[0110] In embodiments of the present invention, the mild replacement solution is prepared using phosphate buffer and a nonionic surfactant. The pH of the phosphate buffer is 7.2–7.4, and the nonionic surfactant is Tween-20, with a volume fraction of 0.01%–0.05% in the mild replacement solution. The phosphate buffer is used to maintain the pH of the liquid environment within the dorsal groove region, reducing changes in the morphology of residual tissue in the dorsal groove and the fluorescence response state of lipid affinity probes during replacement. A low concentration of Tween-20 is used to reduce the nonspecific adhesion of lipid affinity probes that have not formed a stable affinity with the residual tissue in the dorsal groove to the surface of the dorsal groove. This allows the mild replacement solution to gradually replace the original lipid affinity probe solution along the dorsal groove channel after entering the dorsal groove region, carrying away lipid affinity probes in a free or nonspecifically attached state, while retaining lipid affinity probes that have already formed an affinity with the lipid-related residual tissue in the dorsal groove region. Therefore, the lipid affinity probe solution is applied... The first average fluorescence intensity corresponding to the time interval is used to characterize the initial probe loading level in the dorsal groove region before mild replacement. The second average fluorescence intensity corresponding to the time interval of mild replacement solution application is used to characterize the probe loading level remaining in the dorsal groove region after the removal of free and non-specifically attached probes. The retention ratio is obtained by dividing the second average fluorescence intensity by the first average fluorescence intensity, which represents the proportion of probe mass remaining after mild replacement relative to the initial probe mass. When there is more lipid-related residual tissue in the dorsal groove region and a strong retention capacity for lipid-affinity probes, more probes are retained after mild replacement, the second average fluorescence intensity is relatively high, and the retention ratio is large. When there is less lipid-related residual tissue in the dorsal groove region, the proportion of free or non-specifically attached probes removed by the mild replacement solution increases, the second average fluorescence intensity decreases relatively, and the retention ratio is small. Thus, the correspondence between the replacement effect of the mild replacement solution and the retention ratio is established.

[0111] Prepare aqueous wetting solution, lipid affinity probe solution, and mild replacement solution separately. Add each of these solutions to independent reagent containers and connect each container to an independent applicator, ensuring that each applicator corresponds to only one reagent. Perform a drainage process on each applicator to fill the internal channel with reagent and discharge continuous liquid from the applicator outlet. Remove any remaining suspended droplets at the applicator outlet to ensure that the outlet is ready for re-dispensing liquid. Label the applicator corresponding to the aqueous wetting solution as the first applicator, the applicator corresponding to the lipid affinity probe solution as the second applicator, and the applicator corresponding to the mild replacement solution as the third applicator. Arrange the first, second, and third applicators in the order of application of aqueous wetting solution, lipid affinity probe solution, and mild replacement solution. This application order is designated as a three-segment reagent sequence. Record the reagent name, reagent container, applicator, and application order for each reagent in the three-segment reagent sequence.

[0112] Apply the aqueous wetting solution to the inlet point of the dorsal groove of each rice grain, so that the aqueous wetting solution enters the dorsal groove area of ​​the rice grain along the center line of the dorsal groove.

[0113] Move the outlet of the applicator corresponding to the aqueous wetting liquid to above the inlet point of the back groove of the rice grain, align the applicator outlet with the inlet point of the back groove in the image acquisition direction, drive the applicator to release the aqueous wetting liquid, so that the aqueous wetting liquid contacts the inlet point of the back groove, the aqueous wetting liquid forms a liquid contact interface at the inlet point of the back groove, and enters the back groove region defined by the first back groove boundary and the second back groove boundary along the center line of the back groove of the rice grain, and record the moment when the aqueous wetting liquid begins to contact the inlet point of the back groove and the moment when the aqueous wetting liquid stops releasing.

[0114] After the aqueous wetting solution is applied, the lipid affinity probe solution is applied to the dorsal groove inlet point of the same rice grain, so that the lipid affinity probe solution enters the dorsal groove region of the rice grain along the dorsal groove centerline.

[0115] Obtain the dorsal groove inlet point, dorsal groove centerline, first dorsal groove boundary, second dorsal groove boundary, and dorsal groove region corresponding to the rice grain. Move the applicator corresponding to the lipid affinity probe liquid above the dorsal groove inlet point of the rice grain, so that the projection position of the applicator outlet falls on the location of the dorsal groove inlet point. Maintain a gap between the applicator outlet and the rice grain surface that allows liquid contact without pressing the rice grain. Drive the applicator to release the lipid affinity probe liquid, so that the lipid affinity probe liquid forms droplets from the applicator outlet and contacts the dorsal groove inlet point. After contacting the dorsal groove inlet point, the lipid affinity probe liquid enters the dorsal groove region defined by the first and second dorsal groove boundaries. The lipid affinity probe liquid advances in the dorsal groove region along the extension direction of the dorsal groove centerline. Record the moment when the lipid affinity probe liquid begins to contact the dorsal groove inlet point as the start time of the lipid affinity probe liquid application time interval, and record the moment when the applicator stops releasing the lipid affinity probe liquid as the end time of the lipid affinity probe liquid application time interval. Establish a correspondence between the lipid affinity probe liquid application time interval and the rice grain.

[0116] After the lipid affinity probe solution is applied, the mild replacement solution is applied to the dorsal groove inlet point of the same rice grain, so that the mild replacement solution enters the dorsal groove region along the dorsal groove centerline;

[0117] Obtain the inlet point, center line, first boundary, second boundary, and region of the back groove corresponding to the rice grain. Move the applicator corresponding to the mild displacement liquid above the inlet point of the back groove of the rice grain, so that the projection position of the applicator outlet falls on the location of the inlet point. Maintain a gap between the applicator outlet and the surface of the rice grain that allows liquid contact without pressing the rice grain. Drive the applicator to release the mild displacement liquid, so that the mild displacement liquid forms droplets from the applicator outlet and contacts the inlet point of the back groove. After contacting the inlet point, the mild displacement liquid enters the back groove region defined by the first and second back groove boundaries. The mild displacement liquid advances in the back groove region along the extension direction of the center line of the back groove. Record the moment when the mild displacement liquid begins to contact the inlet point of the back groove as the start time of the mild displacement liquid application time interval, and record the moment when the applicator stops releasing the mild displacement liquid as the end time of the mild displacement liquid application time interval. Establish a correspondence between the mild displacement liquid application time interval and the rice grain.

[0118] Record the application time intervals of the aqueous wetting solution, lipid affinity probe solution, and mild replacement solution for the rice grains;

[0119] The application time interval of the aqueous wetting solution refers to the time range from when the aqueous wetting solution begins to enter the back groove region to when the application of the aqueous wetting solution ends; the application time interval of the lipid affinity probe solution refers to the time range from when the lipid affinity probe solution begins to enter the back groove region to when the application of the lipid affinity probe solution ends; the application time interval of the mild replacement solution refers to the time range from when the mild replacement solution begins to enter the back groove region to when the application of the mild replacement solution ends.

[0120] The rice grain identifier, dorsal groove inlet point, dorsal groove region, aqueous wetting solution application time interval, lipid affinity probe solution application time interval, and mild replacement solution application time interval are written into the same application record. The aqueous wetting solution application time interval, lipid affinity probe solution application time interval, and mild replacement solution application time interval are arranged in the actual application sequence so that each time interval corresponds to the same rice grain, the same dorsal groove inlet point, and the same dorsal groove region. The resulting application record is used as the three-segment reagent sequence application record corresponding to the rice grain.

[0121] Based on the application time intervals of the aqueous wetting solution, the lipid affinity probe solution, and the mild replacement solution, a three-segment reagent sequence application record is generated.

[0122] The three-segment reagent sequence application record refers to the data record formed by recording the application time intervals of the aqueous phase wetting solution, the lipid affinity probe solution, and the mild replacement solution in chronological order. This data record is used to characterize the sequential relationship of each reagent entering the dorsal groove region and the corresponding continuous process.

[0123] S3. Based on the three-segment reagent sequence, a recording is applied to obtain the liquid front advancement sequence;

[0124] In an embodiment of the present invention, a liquid front propulsion sequence is obtained, including:

[0125] Within the time intervals of the aqueous wetting solution application, lipid affinity probe solution application, and mild replacement solution application recorded in the three-segment reagent sequence application, continuous bright-field image acquisition was performed on the dorsal groove region to obtain a bright-field image sequence.

[0126] A bright-field image sequence refers to a collection of multiple bright-field images of the back groove region acquired continuously within the time intervals of application of aqueous wetting fluid, lipid affinity probe fluid, and mild replacement fluid, arranged in chronological order of acquisition. This image collection records the dynamic changes in the flow, diffusion, and space occupation of the liquid within the back groove region.

[0127] The clamp holding the rice grain is fixed at the imaging position of the image acquisition device, so that the dorsal groove inlet point, dorsal groove end point, dorsal groove center line, and the dorsal groove region defined by the first and second dorsal groove boundaries of the rice grain are all within the imaging field of view. Bright field illumination is applied to the dorsal groove region of the rice grain, so that the dorsal groove region, liquid-covered region, and uncovered region form a distinguishable brightness distribution in the bright field image. Bright field image acquisition is started when the aqueous wetting liquid begins to contact the dorsal groove inlet point. Images of the dorsal groove region of the rice grain are continuously acquired during the time intervals of aqueous wetting liquid application, lipid affinity probe liquid application, and mild replacement liquid application. A correspondence is established between each acquired bright field image and the corresponding acquisition time. All bright field images of the same rice grain obtained in the three application time intervals are arranged in chronological order of acquisition time to obtain the bright field image sequence of the rice grain.

[0128] Based on the centerline of the back groove, the position of the liquid front in the bright field image sequence is extracted to obtain the liquid front advancement sequence.

[0129] The liquid front position refers to the boundary between the areas reached and unreached by the liquid as it advances within the back trench region. This position reflects the actual range of liquid advancement within the back trench region. Position extraction refers to the process of determining the corresponding position of the liquid front position on the center line of the back trench based on the brightness difference between the liquid-covered and uncovered areas in the brightfield image. The liquid front advancement sequence refers to the set of position data formed by arranging the liquid front positions corresponding to multiple acquisition times in chronological order. This set of data is used to reflect the advancement process and state of the liquid along the center line of the back trench region.

[0130] The center line, inlet point, end point, first and second back groove boundaries of the rice grain are mapped onto each bright-field image in the bright-field image sequence. In each bright-field image, starting from the inlet point, the image brightness distribution within the back groove region defined by the first and second back groove boundaries is examined segment by segment along the center line towards the end point. The continuous area connected to the inlet point and covered by liquid is defined as the liquid-reached area. The area adjacent to the liquid-reached area along the center line but not covered by liquid is defined as the liquid-unreached area. The boundary between the liquid-reached and liquid-unreached areas along the center line is defined as the liquid front position corresponding to that bright-field image. This liquid front position is converted into the arc length position of the back groove center line relative to the inlet point. The arc length position corresponding to each bright-field image is established in correspondence with the acquisition time of that bright-field image and arranged in chronological order of acquisition time to obtain the liquid front advancement sequence of the rice grain.

[0131] S4. Calculate the channel focus based on the central narrow band region and the side band region of the fluorescence image sequence; calculate the membrane retention focus value based on the first average fluorescence intensity, the second average fluorescence intensity, and the channel focus based on the back groove fluorescence intensity sequence.

[0132] In an embodiment of the present invention, calculating the channel focus includes:

[0133] During the time intervals of lipid affinity probe solution application and mild replacement solution application, continuous fluorescence images of the dorsal groove region were acquired. The fluorescence images obtained from each acquisition were arranged in chronological order of acquisition time to obtain a fluorescence image sequence.

[0134] A fluorescence image sequence refers to a collection of multiple fluorescence images of the dorsal groove region acquired continuously within the time intervals of lipid affinity probe solution application and mild replacement solution application, arranged in the order of acquisition time. This image collection records the distribution and change of light signals generated in the dorsal groove region after the lipid affinity probe solution enters the dorsal groove region.

[0135] Fluorescent excitation light is irradiated onto the dorsal groove region of the rice grain, causing the lipid affinity probe solution in the dorsal groove region to generate a fluorescent signal. The imaging field of the fluorescence image acquisition device covers the dorsal groove inlet point, dorsal groove end point, and the dorsal groove region defined by the first and second dorsal groove boundaries of the rice grain. Fluorescent images of the dorsal groove region of the rice grain are continuously acquired during the lipid affinity probe solution application time interval and the mild replacement solution application time interval. The fluorescence images acquired each time are correlated with the acquisition time corresponding to that acquisition. All the obtained fluorescence images are arranged in chronological order of acquisition time to obtain the fluorescence image sequence of the rice grain.

[0136] Based on the dorsal groove centerline, the central narrow band region and lateral band region are determined in the fluorescence image sequence;

[0137] The central narrow band region refers to the band-shaped image region formed with the dorsal sulcus centerline as the center and extending along the dorsal sulcus centerline. This region is used to represent the distribution range of fluorescence signals near the center of the dorsal sulcus. The lateral band region refers to the band-shaped image regions located on both sides of the central narrow band region and distributed along the dorsal sulcus centerline. This region is used to represent the distribution range of fluorescence signals in the adjacent areas on both sides of the dorsal sulcus center.

[0138] The dorsal groove centerline, the first dorsal groove boundary, and the second dorsal groove boundary of the rice grain are mapped onto each fluorescence image in the fluorescence image sequence. Multiple centerline positions are sequentially selected along the dorsal groove centerline from the dorsal groove inlet to the dorsal groove outlet. At each centerline position, the corresponding tangential direction of the dorsal groove centerline is determined. The direction perpendicular to the tangential direction of the dorsal groove centerline and extending to the first and second dorsal groove boundaries is defined as the lateral direction corresponding to that centerline position. The first lateral distance from the centerline position to the first dorsal groove boundary and the second lateral distance from the centerline position to the second dorsal groove boundary are determined along the lateral direction. One-third of the first lateral distance is taken as the first narrow band distance, and one-third of the second lateral distance is taken as the second narrow band distance. Moving the first narrow band distance from the centerline position along the lateral direction towards the first dorsal groove boundary yields the first narrow band boundary position. Moving the second narrow band distance from the centerline position along the lateral direction towards the second dorsal groove boundary yields the second narrow band boundary position. According to the arrangement of the centerline positions along the back groove from the entrance point to the end point, the positions of each first narrow band boundary are connected sequentially to form the first narrow band boundary, and the positions of each second narrow band boundary are connected sequentially to form the second narrow band boundary. The continuous strip-shaped image region enclosed by the first narrow band boundary, the second narrow band boundary, and the end connecting lines corresponding to the entrance point and the end point of the back groove is defined as the central narrow band region. The central narrow band region occupies one-third of the horizontal range between the corresponding first back groove boundary and the second back groove boundary at each horizontal position of the back groove. The image region between the first narrow band boundary and the first back groove boundary, and the image region between the second narrow band boundary and the second back groove boundary are jointly defined as the side band region. Thus, the central narrow band region and the side band region are both limited to the first back groove boundary and the second back groove boundary, and extend continuously along the direction from the entrance point to the end point of the back groove. At the same time, the width of the central narrow band caused by the change in the width of the back groove at different positions can change synchronously with the horizontal range of the corresponding back groove, thereby determining the range of the central narrow band region according to a uniform horizontal ratio.

[0139] Based on the application time interval of the mild replacement fluid, the average fluorescence intensity of the central narrow band region was extracted from the fluorescence image sequence;

[0140] The fluorescence images corresponding to the application time interval of the mild replacement fluid are determined. The central narrow band region in each corresponding fluorescence image is taken as the intensity extraction region. The fluorescence intensity values ​​of all pixels in the central narrow band region are obtained. The fluorescence intensity values ​​of all pixels in the central narrow band region are summed to obtain the sum value of the fluorescence intensity of the central narrow band region. The sum value of the fluorescence intensity of the central narrow band region is divided by the number of pixels in the central narrow band region that participated in the summation to obtain the average fluorescence intensity of the central narrow band region corresponding to that fluorescence image. The average fluorescence intensity of the central narrow band region corresponding to each fluorescence image within the application time interval of the mild replacement fluid is recorded according to the acquisition time, and the recorded result is taken as the average fluorescence intensity of the central narrow band region.

[0141] Based on the application time interval of the mild replacement fluid, the average fluorescence intensity of the sideband region was extracted from the fluorescence image sequence;

[0142] Average fluorescence intensity refers to the average result of the fluorescence intensity values ​​of all pixels in a specified image area. This result is used to represent the overall fluorescence level of the specified image area at the corresponding acquisition time.

[0143] The fluorescence images corresponding to the mild replacement fluid application time intervals are determined. The identified sideband region in each corresponding fluorescence image is used as the intensity extraction region. The fluorescence intensity values ​​of all pixels in the sideband region are obtained. The fluorescence intensity values ​​of all pixels in the sideband region are summed to obtain the sum of the sideband region fluorescence intensity. The sum of the sideband region fluorescence intensity is divided by the number of pixels in the sideband region involved in the summation to obtain the average fluorescence intensity of the sideband region corresponding to that fluorescence image. The average fluorescence intensity of the sideband region corresponding to each fluorescence image within the mild replacement fluid application time interval is recorded according to the acquisition time, and the recorded result is used as the average fluorescence intensity of the sideband region.

[0144] The channel focusing power is obtained by dividing the average fluorescence intensity of the central narrow band region by the average fluorescence intensity of the side band region.

[0145] Channel focus refers to the ratio between the average fluorescence intensity of the central narrow band region and the average fluorescence intensity of the side band region. This ratio is used to indicate the degree to which the fluorescence signal is concentrated along the central direction of the dorsal groove.

[0146] It should be noted that the central narrow band region is located near the center line of the dorsal groove, corresponding to the position where the liquid advances and is retained along the main channel of the dorsal groove. The lateral band region is located on both sides of the center line of the dorsal groove, corresponding to the position where the liquid diffuses and disperses towards the edge of the dorsal groove. When a dorsal groove cryptic membrane exists, the lipid affinity probe solution is more likely to form continuous retention along the main channel of the dorsal groove after the action of a mild replacement solution. Therefore, the fluorescence signal in the central narrow band region shows a higher degree of concentration than the fluorescence signal in the lateral band region. The ratio obtained by dividing the average fluorescence intensity of the central narrow band region by the average fluorescence intensity of the lateral band region can characterize the concentration of the fluorescence signal in the main channel of the dorsal groove and its distribution in the lateral direction of the dorsal groove. The larger the ratio, the more concentrated the fluorescence signal is in the center of the dorsal groove, and the more likely the liquid is to be distributed along the continuous channel in the dorsal groove. The closer the ratio is to one, the smaller the difference in the distribution of the fluorescence signal between the central region and the regions on both sides of the dorsal groove, and the higher the degree of lateral diffusion of the liquid in the dorsal groove. Therefore, the ratio of the average fluorescence intensity of the central narrow band region to the average fluorescence intensity of the lateral band region is used as the channel focusing degree.

[0147] In an embodiment of the present invention, calculating the film retention focusing value includes:

[0148] The fluorescence intensity at the corresponding position of the dorsal groove centerline is extracted frame by frame in the fluorescence image sequence to obtain the dorsal groove fluorescence intensity sequence.

[0149] The dorsal groove fluorescence intensity sequence refers to the set of fluorescence intensity data extracted frame by frame from the fluorescence image sequence along the corresponding position of the dorsal groove center line. This set of data is arranged in the order of acquisition time and is used to represent the light signal change process of the lipid affinity probe solution at the position of the dorsal groove center line.

[0150] The dorsal groove centerline of the rice grain is mapped onto each fluorescence image in the fluorescence image sequence. In each fluorescence image, all image positions corresponding to the dorsal groove centerline are determined along the direction from the dorsal groove inlet point to the dorsal groove end point. The fluorescence intensity value of each image position is obtained. All fluorescence intensity values ​​corresponding to the dorsal groove centerline in the same fluorescence image are summed and divided by the number of image positions involved in the summation to obtain the average fluorescence intensity of the dorsal groove centerline corresponding to that fluorescence image. The average fluorescence intensities of the dorsal groove centerlines corresponding to each fluorescence image are arranged in chronological order of acquisition time to obtain the dorsal groove fluorescence intensity sequence of the rice grain.

[0151] Based on the application time interval of the lipid affinity probe solution, the first average fluorescence intensity was extracted from the dorsal groove fluorescence intensity sequence;

[0152] Based on the three-segment reagent sequence application record of the rice grain, the start and end times of the lipid affinity probe solution application time interval are determined. The average fluorescence intensity of the dorsal groove centerline within the lipid affinity probe solution application time interval is selected from the dorsal groove fluorescence intensity sequence. The selected average fluorescence intensity of the dorsal groove centerline is summed and divided by the number of selected data to obtain the first average fluorescence intensity corresponding to the rice grain.

[0153] Based on the time interval of mild replacement fluid application, the second average fluorescence intensity was extracted from the dorsal groove fluorescence intensity sequence;

[0154] The first average fluorescence intensity refers to the average result of the data corresponding to the dorsal groove fluorescence intensity sequence within the time interval of lipid affinity probe application. This result is used to represent the fluorescence level formed at the center of the dorsal groove after the probe enters the dorsal groove region. The second average fluorescence intensity refers to the average result of the data corresponding to the dorsal groove fluorescence intensity sequence within the time interval of mild replacement solution application. This result is used to represent the fluorescence level still retained at the center of the dorsal groove after the replacement solution has been applied.

[0155] Based on the three-segment reagent sequence application record of the rice grain, the start and end times of the mild replacement solution application time interval are determined. The average fluorescence intensity of the dorsal groove centerline within the mild replacement solution application time interval is selected from the dorsal groove fluorescence intensity sequence. The selected average fluorescence intensity of the dorsal groove centerline is summed and divided by the number of selected data to obtain the second average fluorescence intensity corresponding to the rice grain.

[0156] Divide the second average fluorescence intensity by the first average fluorescence intensity to obtain the retention ratio;

[0157] The retention ratio refers to the quantitative characterization of the ability of lipid-related residual tissue in the dorsal groove region to retain probe substances. It reflects the ability of probe substances to form a binding with residual tissue and maintain their existence after entering the dorsal groove region. When the retention ratio is high, it indicates that there are residual structures in the dorsal groove region that can continuously retain probe substances. When the retention ratio is low, it indicates that probe substances are more likely to detach or migrate from the dorsal groove region.

[0158] It should be noted that the fluorescence intensity originates from the light signal released after the lipid affinity probe is excited. Under the condition that the probe concentration, excitation conditions and collection conditions are kept consistent, the fluorescence intensity is positively correlated with the actual amount of probe material retained in the unit area. Therefore, the first average fluorescence intensity can characterize the initial probe loading level formed after the lipid affinity probe solution enters the dorsal groove region, and the second average fluorescence intensity can characterize the probe loading level still retained in the dorsal groove region after the action of a mild replacement solution. According to the mass retention relationship, the ratio between the residual probe mass and the initial probe mass can characterize the degree of probe material retention. Since the fluorescence intensity corresponds to the probe mass, the ratio of the second average fluorescence intensity to the first average fluorescence intensity is used as the retention ratio. This can eliminate the influence of the difference in the initial probe entry amount between different rice grains, so that the calculation results directly reflect the relative retention capacity of the dorsal groove region for the probe material.

[0159] Multiply the retention ratio by the channel focusing degree to obtain the membrane retention focusing value.

[0160] The membrane retention focusing value refers to the comprehensive characterization of the retention state of probe material on residual tissue in the dorsal groove region and its concentrated distribution along the main channel of the dorsal groove. It reflects the continuity, adhesion and spatial distribution of residual tissue in the dorsal groove region. When the membrane retention focusing value is high, it indicates that there is residual tissue in the dorsal groove region that is continuously distributed along the main channel of the dorsal groove and can stably retain probe material. When the membrane retention focusing value is low, it indicates that the continuity of residual tissue is reduced, the distribution tends to be more discrete or the ability to retain probe material is weakened.

[0161] It should be noted that channel focusing degree reflects the spatial concentration of probe material in the main channel of the dorsal groove, while retention ratio reflects the retention degree of probe material in the dorsal groove region. These two correspond to two independent dimensions: the spatial distribution state and the retention state of the probe material. According to the principle of constructing a joint characterization quantity in physics, when a target state is simultaneously affected by both the retention degree and the concentration degree, a comprehensive characterization quantity can be constructed by multiplying the dimensionless ratios. This ensures that the comprehensive result decreases synchronously when either dimension decreases. The comprehensive result only increases when the probe material can be stably retained on the residual tissue and form a continuous concentrated distribution along the main channel of the dorsal groove. Therefore, multiplying the retention ratio by the channel focusing degree yields the membrane retention focusing value. This result can simultaneously reflect the adhesion and retention ability of the residual tissue to the probe material and the continuous distribution state of the residual tissue along the main channel of the dorsal groove, thus characterizing the overall existence state of the residual tissue in the dorsal groove region.

[0162] S5. Based on the membrane retention focusing value and the liquid front propulsion sequence, the accuracy of the rice sample to be tested is judged, and the judgment result is obtained.

[0163] In embodiments of the present invention, obtaining the judgment result includes:

[0164] Based on the application time interval of the lipid affinity probe solution, the maximum liquid front advancement length was extracted from the liquid front advancement sequence;

[0165] The maximum liquid front advance length refers to the farthest arc length reached by the liquid along the dorsal groove centerline from the dorsal groove inlet point to the dorsal groove end point within the time interval of lipid affinity probe liquid application. This length reflects the actual entry range of the lipid affinity probe liquid in the dorsal groove region.

[0166] The three-segment reagent sequence application record and liquid front advancement sequence corresponding to the rice grain were obtained. The three-segment reagent sequence application record includes the start and end times of the lipid affinity probe solution application time interval. The liquid front advancement sequence includes multiple acquisition times and the liquid front position corresponding to each acquisition time. The liquid front position that is located within the lipid affinity probe solution application time interval of the acquisition time is selected. The selected liquid front position is compared with the arc length of the corresponding dorsal groove centerline. The arc length of the dorsal groove centerline that is farthest from the dorsal groove entrance point is determined as the maximum liquid front advancement length of the rice grain.

[0167] The propulsion occupancy ratio is calculated based on the maximum liquid front propulsion length and the length of the back groove centerline;

[0168] The length of the dorsal groove centerline refers to the total length measured along the path extending from the dorsal groove inlet to the dorsal groove end. This length reflects the complete path range available for liquid to advance along the main channel in the dorsal groove region of the rice grain. The advance occupancy ratio refers to the proportion of the maximum liquid front advance length relative to the length of the dorsal groove centerline. This proportion reflects the coverage of the lipid affinity probe liquid in the main channel of the dorsal groove.

[0169] Obtain the maximum liquid front propulsion length and the length of the back groove centerline corresponding to the grain. The length of the back groove centerline is the total arc length measured along the back groove centerline from the back groove inlet point to the back groove end point of the grain. Divide the maximum liquid front propulsion length by the length of the back groove centerline to obtain the propulsion occupancy ratio of the grain.

[0170] The propulsion occupancy ratio is characterized by the ratio of the maximum liquid front propulsion length to the dorsal groove centerline length. The maximum liquid front propulsion length represents the furthest position the lipid affinity probe fluid can actually reach within the dorsal groove region, corresponding to the actual area occupied by the fluid in the main channel of the dorsal groove. The dorsal groove centerline length represents the complete path length along the main channel from the dorsal groove inlet to the dorsal groove outlet, corresponding to the theoretically complete channel range that the fluid can cover. Since both have the same length dimension, the dimensionless ratio is obtained by dividing the maximum liquid front propulsion length by the dorsal groove centerline length. This ratio represents the liquid... The ratio of the actual length occupied by the liquid to the total length of the main channel of the back groove is determined by the following: when the liquid reaches the end of the back groove, the maximum liquid front advancement length is equal to the length of the back groove centerline, and the resulting ratio is close to the complete occupancy state. When the liquid can only reach a part of the back groove centerline, the maximum liquid front advancement length is less than the length of the back groove centerline, and the resulting ratio corresponds to the partial occupancy state. Therefore, using the ratio of the maximum liquid front advancement length to the length of the back groove centerline as the advancement occupancy ratio can eliminate the influence of the difference in back groove length between different grains and directly characterize the actual degree of liquid occupancy of the main channel of the back groove.

[0171] Based on the membrane retention focus value and the propulsion occupancy ratio, the processing accuracy assessment value is calculated;

[0172] The processing accuracy assessment value refers to the quantitative result formed by the membrane retention focus value and the propulsion occupancy ratio, which is used to characterize the state of the dorsal groove residual tissue. This result reflects the comprehensive relationship between the dorsal groove residual tissue in the batch of rice samples to be tested and the retention, concentrated distribution and liquid entry range of the probe material.

[0173] Specifically, the membrane retention focus value and propulsion occupancy ratio are obtained for each grain in the set of rice grains to be tested. The same grain corresponds to multiple membrane retention focus values ​​at different acquisition times. These values ​​are arranged in chronological order according to the acquisition times of the fluorescence image sequence to form a membrane retention focus value sequence for that grain. The propulsion occupancy ratio is a single value calculated based on the maximum liquid front propulsion length and the length of the dorsal groove centerline corresponding to that grain. For each grain in the set of rice grains to be tested, all membrane retention focus values ​​are extracted from the membrane retention focus value sequence corresponding to that grain. All extracted membrane retention focusing values ​​are summed to obtain the sum of membrane retention focusing values ​​for each grain of rice. The number of membrane retention focusing values ​​participating in the summation in the sequence of membrane retention focusing values ​​for each grain of rice is determined. The sum of membrane retention focusing values ​​is divided by the number of membrane retention focusing values ​​to obtain the single-grain membrane retention focusing value for each grain of rice. The single-grain membrane retention focusing value is a single value obtained by summarizing the membrane retention focusing states of the same grain of rice at multiple acquisition times. This transforms the original sequence of membrane retention focusing values ​​for the same grain of rice, arranged according to the acquisition time, into a single membrane retention focusing result that can participate in the calculation of single-grain processing accuracy.

[0174] Obtain the propulsion occupancy ratio corresponding to each grain of rice. Divide the single-grain membrane retention focus value corresponding to each grain of rice by the propulsion occupancy ratio to obtain the single-grain processing accuracy response value corresponding to each grain of rice. Each grain of rice corresponds to only one single-grain membrane retention focus value, one propulsion occupancy ratio, and one single-grain processing accuracy response value. The single-grain processing accuracy response value is no longer sequenced according to the acquisition time of the fluorescence image sequence. Following the above method, summarize the membrane retention focus value sequences corresponding to each grain of rice in the test grain set, and calculate the single-grain processing accuracy response value corresponding to each grain of rice in the test grain set by combining the propulsion occupancy ratio of the corresponding grain. Establish a one-to-one correspondence between each single-grain processing accuracy response value and the corresponding grain of rice.

[0175] The number of rice grains in the set of rice grains to be tested is obtained. The single-grain processing accuracy response values ​​corresponding to all rice grains in the set of rice grains to be tested are summed to obtain the single-grain processing accuracy response value sum. The single-grain processing accuracy response value sum is divided by the number of rice grains in the set of rice grains to be tested to obtain the processing accuracy identification value of the batch of rice samples to be tested. Thus, the membrane retention focusing value sequence is first summarized within the range of a single rice grain to form a single single-grain membrane retention focusing value. Then, combined with the propulsion occupancy ratio corresponding to that rice grain, a single single-grain processing accuracy response value is obtained. Finally, the single-grain processing accuracy response values ​​corresponding to multiple rice grains are summarized so that the processing accuracy identification value corresponds to the batch of rice samples to be tested.

[0176] Based on the processing precision identification value, the precision of the rice sample batch to be tested is judged, and the judgment result is obtained.

[0177] Accuracy judgment refers to the inspection process of determining the degree of processing of the rice sample to be tested based on the processing accuracy identification value; the judgment result refers to the conclusion on the processing accuracy of the rice sample to be tested obtained through accuracy judgment, which is used to indicate the processing status of the rice sample to be tested.

[0178] Specifically, the processing precision identification value corresponding to the batch of rice to be tested is obtained, and a set of processing precision identification values ​​for standard samples is established. Each processing precision identification value in the set of standard sample processing precision identification values ​​is associated with a known processing precision level of the corresponding standard sample. The set of standard sample processing precision identification values ​​is sorted according to the order of processing precision levels. The processing precision identification value corresponding to the batch of rice to be tested is compared sequentially with each processing precision identification value in the set of standard sample processing precision identification values ​​to determine the positional relationship of the processing precision identification value corresponding to the batch of rice to be tested within the set of standard sample processing precision identification values. When a certain processing precision assessment value in the set of fixed values ​​is matched, the processing precision level corresponding to that value is determined as the processing precision level of the rice sample to be tested. When the processing precision assessment value of the rice sample to be tested is located between the processing precision assessment values ​​of two adjacent standard samples, the processing precision level corresponding to those two adjacent standard sample processing precision assessment values ​​is determined. Based on the closeness between the processing precision assessment value of the rice sample to be tested and those two adjacent standard sample processing precision assessment values, the processing precision level of the rice sample to be tested is determined. The determined processing precision level is then associated with the rice sample to be tested, and the processing precision level of the rice sample to be tested is output as the judgment result.

[0179] The above description is only a preferred embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any equivalent substitutions or modifications made by those skilled in the art within the scope of the technology disclosed in the present invention, based on the technical solution and inventive concept of the present invention, should be covered within the scope of protection of the present invention.

Claims

1. A method for determining the processing precision of rice, characterized in that, Includes the following steps: S1. Determine the dorsal groove region of the rice grain based on the bright-field image of the dorsal groove of the rice grain. S2. Based on the three-segment reagent sequence and the dorsal groove region of the rice grain, generate a three-segment reagent sequence and apply it for recording; S3. Based on the three-segment reagent sequence, a recording is applied to obtain the liquid front advancement sequence; S4. Calculate the channel focus based on the central narrow band region and the side band region of the fluorescence image sequence; The membrane retention focusing value is calculated based on the first average fluorescence intensity, the second average fluorescence intensity, and the channel focusing degree of the back groove fluorescence intensity sequence. S5. Based on the membrane retention focusing value and the liquid front propulsion sequence, the accuracy of the rice sample to be tested is judged, and the judgment result is obtained.

2. The method for determining the processing precision of rice according to claim 1, characterized in that, Determine the dorsal groove region of the rice grain, including: Obtain the set of rice grains to be tested from the rice sample of the batch to be tested; Acquire a bright-field image of the back groove of each rice grain in the set of rice grains to be tested; In the bright-field image of the back gutter, determine the inlet point, end point, and centerline of the back gutter; Determine the boundary of the back groove in the bright-field image of the back groove; The back groove region of the rice grain is determined based on the inlet point, end point, center line, and boundary of the back groove.

3. The method for determining the processing precision of rice according to claim 2, characterized in that, Generate a three-segment reagent sequence and apply the record, including: A three-stage reagent sequence was configured; wherein the three-stage reagent sequence includes an aqueous wetting solution, a lipid affinity probe solution, and a mild replacement solution; Apply the aqueous wetting solution to the inlet point of the dorsal groove of each rice grain, so that the aqueous wetting solution enters the dorsal groove area of ​​the rice grain along the center line of the dorsal groove. After the aqueous wetting solution is applied, the lipid affinity probe solution is applied to the dorsal groove inlet point of the same rice grain, so that the lipid affinity probe solution enters the dorsal groove region of the rice grain along the dorsal groove centerline. After the lipid affinity probe solution is applied, the mild replacement solution is applied to the dorsal groove inlet point of the same rice grain, so that the mild replacement solution enters the dorsal groove region along the dorsal groove centerline; Record the application time intervals of the aqueous wetting solution, lipid affinity probe solution, and mild replacement solution for the rice grains; Based on the application time intervals of the aqueous wetting solution, the lipid affinity probe solution, and the mild replacement solution, a three-segment reagent sequence application record is generated.

4. The method for determining the processing precision of rice according to claim 2, characterized in that, The liquid front propulsion sequence was obtained, including: Within the time intervals of the aqueous wetting solution application, the lipid affinity probe solution application, and the mild replacement solution application recorded in the three-segment reagent sequence application, continuous bright-field image acquisition was performed on the dorsal groove region to obtain a bright-field image sequence. Based on the centerline of the back groove, the position of the liquid front in the bright field image sequence is extracted to obtain the liquid front advancement sequence.

5. The method for determining the processing precision of rice according to claim 4, characterized in that, Calculating channel focus includes: During the time intervals of lipid affinity probe solution application and mild replacement solution application, continuous fluorescence images of the dorsal groove region were acquired. The fluorescence images obtained from each acquisition were arranged in chronological order of acquisition time to obtain a fluorescence image sequence. Based on the dorsal groove centerline, the central narrow band region and lateral band region are determined in the fluorescence image sequence; Based on the application time interval of the mild replacement fluid, the average fluorescence intensity of the central narrow band region was extracted from the fluorescence image sequence; Based on the application time interval of the mild replacement fluid, the average fluorescence intensity of the sideband region was extracted from the fluorescence image sequence; The channel focusing power is obtained by dividing the average fluorescence intensity of the central narrow band region by the average fluorescence intensity of the side band region.

6. The method for determining the processing precision of rice according to claim 5, characterized in that, Calculating the film retention focus value includes: The fluorescence intensity at the corresponding position of the dorsal groove centerline is extracted frame by frame in the fluorescence image sequence to obtain the dorsal groove fluorescence intensity sequence. Based on the application time interval of the lipid affinity probe solution, the first average fluorescence intensity was extracted from the dorsal groove fluorescence intensity sequence; Based on the time interval of mild replacement fluid application, the second average fluorescence intensity was extracted from the dorsal groove fluorescence intensity sequence; Divide the second average fluorescence intensity by the first average fluorescence intensity to obtain the retention ratio; Multiply the retention ratio by the channel focusing degree to obtain the membrane retention focusing value.

7. The method for determining the processing precision of rice according to claim 3, characterized in that, The judgment result includes: Based on the application time interval of the lipid affinity probe solution, the maximum liquid front advancement length was extracted from the liquid front advancement sequence; The propulsion occupancy ratio is calculated based on the maximum liquid front propulsion length and the length of the back groove centerline; Based on the membrane retention focus value and the propulsion occupancy ratio, the processing accuracy assessment value is calculated; Based on the processing precision identification value, the precision of the rice sample batch to be tested is judged, and the judgment result is obtained.

8. The method for determining the processing precision of rice according to claim 2, characterized in that, In the bright-field image of the back gutter, determine the inlet point, end point, and centerline of the back gutter, including: Extract the outer contour of rice grains from the bright-field image of the back ditch; Based on the outer contour of the rice grain, extract the dark groove area on the back; Connectivity screening is performed on the dark texture region of the back groove to obtain candidate regions for the back groove; The candidate regions of the back groove are skeletonized to obtain the candidate skeleton lines of the back groove. One end of the candidate skeleton line of the back groove is determined as the entry point of the back groove, and the other end of the candidate skeleton line of the back groove is determined as the end point of the back groove. The candidate skeleton line of the backditch connecting the inlet point and the end point of the backditch is determined as the center line of the backditch.

9. The method for determining the processing precision of rice according to claim 2, characterized in that, Determining the back groove boundary in a bright-field image includes: Along the direction perpendicular to the center line of the back groove, the dark texture area of ​​the back groove corresponding to the bright field image of the back groove is scanned point by point to obtain multiple horizontal grayscale distributions. In each horizontal grayscale distribution, determine the grayscale change positions on both sides of the back groove center line; Connecting multiple grayscale change locations on one side of the back groove centerline yields the first back groove boundary; Connecting multiple grayscale change locations on the other side of the back groove centerline yields the second back groove boundary; The first back groove boundary and the second back groove boundary are defined as back groove boundaries.

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