CN109932517B - Blood agglutination judgment method and device - Google Patents
Blood agglutination judgment method and device Download PDFInfo
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- CN109932517B CN109932517B CN201910268346.2A CN201910268346A CN109932517B CN 109932517 B CN109932517 B CN 109932517B CN 201910268346 A CN201910268346 A CN 201910268346A CN 109932517 B CN109932517 B CN 109932517B
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Abstract
The blood agglutination judging method and device, which places the blood cell or serum of known blood type in the sample hole of the microporous plate in advance, and places the blood cell or serum of the detected blood in each sample hole to form a reaction sample; scanning a reaction sample in the sample hole to obtain an absorbance value of the light absorption point, defining an effective light absorption point range by taking a middle light absorption point as a center, and removing the light absorption points with errors at the edges; selecting a minimum absorbance value and a maximum absorbance value within the range of the effective light absorption points; taking the minimum absorbance value as a base number, acquiring a difference value between the maximum absorbance value and the minimum absorbance value, and calculating a ratio of the difference value to the minimum absorbance value as the base number; presetting an agglutination reference value, comparing the ratio with the agglutination reference value, judging the blood in the sample hole to be agglutinated when the ratio is greater than the agglutination reference value, and judging the blood in the sample hole to be unagglutinated when the ratio is less than or equal to the agglutination reference value. Whether the mixed sample is agglutinated or not is judged quickly, the accuracy is high, and the application range is wide.
Description
Technical Field
The invention relates to a blood agglutination judgment method and device, and relates to the technical field of blood treatment.
Background
It is known that blood agglutination is a possible serum immune reaction when blood of different blood types meet, and under a microscope, blood cells in the blood are adhered to each other to form a cluster. The microplate readers have the highest popularization rate in various blood stations, hospitals and various blood laboratories, and have a scanning function.
At present, the blood coagulation judgment methods include the following methods: one is an image analysis method, which judges whether blood is coagulated by capturing an image and analyzing the image, and is generally used in large-scale equipment such as a fully automatic blood coagulation analyzer, a blood typing instrument, and a biochemical analyzer. The other is by visual judgment, such as an ABO blood group test card. The first mode adopts large-scale equipment, and cannot be used due to high cost and low popularization rate; the second mode adopts manual work to carry out naked eyes, and detection efficiency is low, and the accuracy of testing result is poor.
Disclosure of Invention
The invention aims to provide a blood agglutination judging method and device, which can quickly judge whether the mixed sample in a sample hole is agglutinated or not, and have high detection accuracy and wide application range.
The technical scheme for solving the technical problems is as follows: a blood coagulation judging method comprising the steps of:
11) placing blood cells or serum of known blood types in sample holes of a microporous plate in advance, and then placing the blood cells or serum of detected blood in each sample hole to form a reaction sample;
12) scanning the reaction sample in the sample hole to obtain the absorbance values of a plurality of light absorption points, defining the range of the effective light absorption points by taking the middle light absorption point as the center, and removing the light absorption points with errors at the edges;
13) selecting a minimum absorbance value and a maximum absorbance value within the effective light absorption point range;
14) taking the minimum absorbance value as a base number, acquiring a difference value between the maximum absorbance value and the minimum absorbance value, and calculating a ratio of the difference value to the minimum absorbance value as the base number;
15) presetting an agglutination reference value, comparing the ratio with the agglutination reference value, judging the blood in the sample hole to be agglutinated when the ratio is larger than the agglutination reference value, and judging the blood in the sample hole to be not agglutinated when the ratio is smaller than or equal to the agglutination reference value.
As a preferred scheme of the blood agglutination judging method, after blood cells or serum of a known blood type and blood cells or serum of detected blood are placed in a sample hole and mixed, reaction time of a preset time period is given, and after the reaction time is up, absorbance scanning is carried out on the mixed reaction sample.
Preferably, the method for determining blood agglutination is performed by scanning absorbance values of the reaction sample in the sample well with a spectrophotometer.
As a preferable scheme of the blood agglutination determination method, the effective light absorption point range is a circular area with a center point as a circle center and a radius of r, when the light absorption point is located within the circular area range, the light absorption point is used as an effective light absorption point, when the light absorption point is located outside the circular area range, the light absorption point is used as an error light absorption point, and the absorbance value of the error light absorption point is removed.
As a preferable mode of the blood agglutination determination method, the agglutination reference value is determined by:
21) placing corpuscles or serum of known blood types in sample holes of a microporous plate in advance, and then placing corpuscles or serum of blood of given blood types in each sample hole to form a reaction sample;
22) scanning the reaction sample of the sample hole to obtain the absorbance values of a plurality of light absorption points, defining the range of the effective light absorption points by taking the middle light absorption point as the center, and removing the light absorption points with errors at the edges;
23) selecting a minimum absorbance value and a maximum absorbance value within the effective light absorption point range;
24) taking the minimum absorbance value as a base number, acquiring a difference value between the maximum absorbance value and the minimum absorbance value, and calculating a ratio of the difference value to the minimum absorbance value as the base number;
25) repeating the steps 21) to 24) n times, and taking the average value of the n-times ratio as an agglutination reference value.
An embodiment of the present invention further provides a blood coagulation determination device, including:
the micro-porous plate is used for placing blood cells or serum of known blood types in sample holes of the micro-porous plate in advance, and then placing the blood cells or serum of detected blood in each sample hole to form a reaction sample;
the light absorption point scanning module is used for scanning the reaction sample in the sample hole to obtain the absorbance values of a plurality of light absorption points;
the effective range definition module is used for defining the effective light absorption point range by taking the middle light absorption point as a center;
the error point removing module is used for removing the light absorbing points with errors on the edges;
the numerical value selection module is used for selecting a minimum absorbance value and a maximum absorbance value within the effective light absorption point range;
the ratio acquisition module is used for acquiring a difference value between the maximum absorbance value and the minimum absorbance value by taking the minimum absorbance value as a base number, and calculating the ratio of the difference value to the minimum absorbance value as the base number;
the reference value presetting module is used for presetting an agglutination reference value;
and the agglutination judgment module is used for comparing the ratio with the agglutination reference value, judging the blood in the sample hole to be agglutinated when the ratio is greater than the agglutination reference value, and judging the blood in the sample hole to be not agglutinated when the ratio is less than or equal to the agglutination reference value.
The blood agglutination judging device further comprises a reaction time setting module, wherein the reaction time setting module is used for setting the reaction time of a preset time period after the blood cells or the serum with known blood types and the blood cells or the serum of the detected blood are placed in the sample hole and mixed, and performing absorbance scanning on the mixed reaction sample after the reaction time is reached.
As a preferable scheme of the blood agglutination judging device, the light absorption point scanning module scans and acquires the absorbance value of the reaction sample in the sample hole through a spectrophotometer.
As a preferable embodiment of the blood coagulation determining device, in the effective range defining module, the effective light absorption point range is a circular area with a center point as a circle center and a radius of r, when the light absorption point is located within the circular area range, the light absorption point is used as an effective light absorption point, when the light absorption point is located outside the circular area range, the light absorption point is used as an error light absorption point, and an absorbance value of the error light absorption point is rejected by the error point rejecting module.
The invention has the beneficial effects that: by taking the middle light absorption point as a center and defining the range of the effective light absorption point, the light absorption points with possible errors at the edge are removed; selecting the minimum absorbance as a base number in the absorbance values in the effective range; selecting the maximum absorbance from the absorbance values in the effective range; obtaining the ratio of the difference value to the minimum absorbance according to the difference value of the maximum absorbance and the minimum absorbance; this ratio is compared with a preset value, above which agglutination is considered to be present. The technical scheme of the embodiment of the invention is simple, has high accuracy, is convenient to integrate into a data analysis system of each blood detection laboratory, can fully utilize the resources of the existing microplate reader, and realizes the rapid and accurate judgment of blood type agglutination.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below. It should be apparent that the drawings in the following description are merely exemplary, and that other embodiments can be derived from the drawings provided by those of ordinary skill in the art without inventive effort.
FIG. 1 is a schematic flow chart of a method for determining blood coagulation according to an embodiment of the present invention;
FIG. 2 is a schematic diagram illustrating a flow of agglutination reference value determination in the blood agglutination determination method according to the embodiment of the present invention;
fig. 3 is a schematic view of a blood coagulation analyzer according to an embodiment of the present invention.
Detailed Description
In order to make the aforementioned objects, features and advantages of the present invention comprehensible, embodiments accompanied with figures are described in detail below. In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present invention. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used herein in the description of the invention is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention.
As shown in fig. 1, an embodiment of the present invention provides a method for determining blood coagulation, including the following steps:
s11: placing blood cells or serum of known blood types in sample holes of a microporous plate in advance, and then placing the blood cells or serum of detected blood in each sample hole to form a reaction sample;
s12: scanning the reaction sample in the sample hole to obtain the absorbance values of a plurality of light absorption points, defining the range of the effective light absorption points by taking the middle light absorption point as the center, and removing the light absorption points with errors at the edges;
s13: selecting a minimum absorbance value and a maximum absorbance value within the effective light absorption point range;
s14: taking the minimum absorbance value as a base number, acquiring a difference value between the maximum absorbance value and the minimum absorbance value, and calculating a ratio of the difference value to the minimum absorbance value as the base number;
s15: presetting an agglutination reference value, comparing the ratio with the agglutination reference value, judging the blood in the sample hole to be agglutinated when the ratio is larger than the agglutination reference value, and judging the blood in the sample hole to be not agglutinated when the ratio is smaller than or equal to the agglutination reference value.
In an embodiment of the method for determining blood agglutination, after the corpuscles or serum of the known blood type are placed in the sample holes and mixed with the corpuscles or serum of the blood to be detected, a reaction time of a preset time period is given, and after the reaction time is reached, an absorbance scan is performed on the mixed reaction sample. When abnormal blood meets each other, the serum agglutinin in one blood can cause the corresponding agglutinogens on the surfaces of blood cells in the other blood to generate immune reaction, and the blood cells in the blood can be adhered and agglomerated under a microscope. And thus can be used to determine blood type by blood coagulation.
In one embodiment of the method for determining agglutination of blood, the absorbance value of the reaction sample in the sample hole is scanned and obtained by a spectrophotometer. The spectrophotometer can decompose the light with complex components into spectral lines, and light waves emitted by a light source lamp of the spectrophotometer are changed into a beam of monochromatic light through a light filter or a monochromator and enter a reaction sample in the micropore pole. One part of the monochromatic light is absorbed by the reaction sample, the other part of the monochromatic light penetrates through the reaction sample and irradiates on a photoelectric detector, and the photoelectric detector converts different light signals with different intensities of the reaction sample to be detected into corresponding electric signals. The absorbance of the reaction sample is different according to whether blood agglutination occurs, and whether blood agglutination occurs can be judged according to the absorbance. Specifically, the effective light absorption point range is a circular area with a center point as a circle center and a radius of r, when the light absorption point is located in the circular area range, the light absorption point is used as the effective light absorption point, when the light absorption point is located outside the circular area range, the light absorption point is used as an error light absorption point, and the absorbance value of the error light absorption point is eliminated.
Referring to fig. 2, in an embodiment of the blood agglutination determination method, the agglutination reference value is determined by:
s21: placing corpuscles or serum of known blood types in sample holes of a microporous plate in advance, and then placing corpuscles or serum of blood of given blood types in each sample hole to form a reaction sample;
s22: scanning the reaction sample of the sample hole to obtain the absorbance values of a plurality of light absorption points, defining the range of the effective light absorption points by taking the middle light absorption point as the center, and removing the light absorption points with errors at the edges;
s23: selecting a minimum absorbance value and a maximum absorbance value within the effective light absorption point range;
s24: taking the minimum absorbance value as a base number, acquiring a difference value between the maximum absorbance value and the minimum absorbance value, and calculating a ratio of the difference value to the minimum absorbance value as the base number;
s25: repeating the steps 21) to 24) n times, and taking the average value of the n-times ratio as an agglutination reference value.
Referring to fig. 3, an embodiment of the present invention further provides a blood coagulation judging device, including:
the micro-porous plate 1 is used for placing blood cells or serum of known blood types in sample holes of the micro-porous plate in advance, and then placing the blood cells or serum of detected blood in each sample hole to form a reaction sample;
the light absorption point scanning module 2 is used for scanning the reaction sample in the sample hole to obtain the absorbance values of a plurality of light absorption points;
the effective range defining module 3 is used for defining the effective light absorption point range by taking the middle light absorption point as a center;
the error point removing module 4 is used for removing the light absorbing points with errors on the edges;
the numerical value selection module 5 is used for selecting a minimum absorbance value and a maximum absorbance value within the effective light absorption point range;
a ratio obtaining module 6, configured to obtain a difference between the maximum absorbance and the minimum absorbance using the minimum absorbance as a base, and calculate a ratio between the difference and the minimum absorbance as the base;
a reference value presetting module 7 for presetting an agglutination reference value;
and the agglutination judgment module 8 is used for comparing the ratio with the agglutination reference value, judging the blood in the sample hole to be agglutinated when the ratio is greater than the agglutination reference value, and judging the blood in the sample hole to be not agglutinated when the ratio is less than or equal to the agglutination reference value.
In an embodiment of the blood agglutination determination apparatus, the apparatus further includes a reaction time setting module 9, configured to set a reaction time of a preset time period after blood cells or serum of a known blood type are placed in the sample hole and mixed with blood cells or serum of the blood to be detected, and perform absorbance scanning on the mixed reaction sample after the reaction time is reached. And the light absorption point scanning module 2 scans and acquires the absorbance value of the reaction sample in the sample hole through a spectrophotometer.
In an embodiment of the blood coagulation determining apparatus, in the effective range defining module 3, an effective light absorption point range is a circular area with a center point as a circle center radius r, when the light absorption point is located within the circular area range, the light absorption point is used as an effective light absorption point, when the light absorption point is located outside the circular area range, the light absorption point is used as an error light absorption point, and an absorbance value of the error light absorption point is rejected by the error point rejecting module 4.
The embodiment of the invention is characterized in that the corpuscles or the serum with known blood types are placed in the sample holes of the microporous plate in advance, and then the corpuscles or the serum of the detected blood is placed in each sample hole to form a reaction sample; scanning a reaction sample in the sample hole to obtain absorbance values of a plurality of light absorption points, defining the range of effective light absorption points by taking a middle light absorption point as a center, and removing the light absorption points with errors at the edges; selecting a minimum absorbance value and a maximum absorbance value within the range of the effective light absorption points; taking the minimum absorbance value as a base number, acquiring a difference value between the maximum absorbance value and the minimum absorbance value, and calculating a ratio of the difference value to the minimum absorbance value as the base number; presetting an agglutination reference value, comparing the ratio with the agglutination reference value, judging the blood in the sample hole to be agglutinated when the ratio is greater than the agglutination reference value, and judging the blood in the sample hole to be unagglutinated when the ratio is less than or equal to the agglutination reference value. The embodiment of the invention takes the middle light absorption point as the center, and eliminates the light absorption points with possible errors at the edge by defining the range of the effective light absorption points; selecting the minimum absorbance as a base number in the absorbance values in the effective range; selecting the maximum absorbance from the absorbance values in the effective range; obtaining the ratio of the difference value to the minimum absorbance according to the difference value of the maximum absorbance and the minimum absorbance; this ratio is compared with a preset value, above which agglutination is considered to be present. The technical scheme of the embodiment of the invention is simple, has high accuracy, is convenient to integrate into a data analysis system of each blood detection laboratory, can fully utilize the resources of the existing microplate reader, and realizes the rapid and accurate judgment of blood type agglutination.
The technical features of the embodiments described above may be arbitrarily combined, and for the sake of brevity, all possible combinations of the technical features in the embodiments described above are not described, but should be considered as being within the scope of the present specification as long as there is no contradiction between the combinations of the technical features.
The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.
Claims (8)
1. A method for determining blood coagulation, comprising the steps of:
11) placing blood cells or serum of known blood types in sample holes of a microporous plate in advance, and then placing the blood cells or serum of detected blood in each sample hole to form a reaction sample;
12) scanning a reaction sample in the sample hole to obtain absorbance values of a plurality of light absorption points, defining an effective light absorption point range by taking a middle light absorption point as a center, and removing light absorption points with errors at the edge, wherein the effective light absorption point range is a circular area with the middle point as a circle center and the radius of r, when the light absorption points are positioned in the circular area range, the light absorption points are taken as the effective light absorption points, when the light absorption points are positioned outside the circular area range, the light absorption points are taken as the error light absorption points, and the absorbance values of the error light absorption points are removed;
13) selecting a minimum absorbance value and a maximum absorbance value within the effective light absorption point range;
14) taking the minimum absorbance value as a base number, acquiring a difference value between the maximum absorbance value and the minimum absorbance value, and calculating a ratio of the difference value to the minimum absorbance value as the base number;
15) presetting an agglutination reference value, comparing the ratio with the agglutination reference value, judging the blood in the sample hole to be agglutinated when the ratio is larger than the agglutination reference value, and judging the blood in the sample hole to be not agglutinated when the ratio is smaller than or equal to the agglutination reference value.
2. A method for determining blood coagulation according to claim 1, wherein a reaction time is given for a predetermined period of time after the blood cells or serum of the known blood type and the blood cells or serum of the blood to be detected are mixed in the sample well, and an absorbance scan is performed on the mixed reaction sample after the reaction time is reached.
3. The method of claim 1, wherein the absorbance value of the reaction sample in the sample well is scanned by a spectrophotometer.
4. A method for determining blood coagulation according to claim 1, wherein said coagulation reference value is determined by:
21) placing corpuscles or serum of known blood types in sample holes of a microporous plate in advance, and then placing corpuscles or serum of blood of given blood types in each sample hole to form a reaction sample;
22) scanning the reaction sample of the sample hole to obtain the absorbance values of a plurality of light absorption points, defining the range of the effective light absorption points by taking the middle light absorption point as the center, and removing the light absorption points with errors at the edges;
23) selecting a minimum absorbance value and a maximum absorbance value within the effective light absorption point range;
24) taking the minimum absorbance value as a base number, acquiring a difference value between the maximum absorbance value and the minimum absorbance value, and calculating a ratio of the difference value to the minimum absorbance value as the base number;
25) repeating the steps 21) to 24) n times, and taking the average value of the n-times ratio as an agglutination reference value.
5. A blood coagulation judging device characterized by comprising:
the micro-porous plate is used for placing blood cells or serum of known blood types in sample holes of the micro-porous plate in advance, and then placing the blood cells or serum of detected blood in each sample hole to form a reaction sample;
the light absorption point scanning module is used for scanning the reaction sample in the sample hole to obtain the absorbance values of a plurality of light absorption points;
the effective range definition module is used for defining the effective light absorption point range by taking the middle light absorption point as a center;
the error point removing module is used for removing the light absorbing points with errors on the edges;
the numerical value selection module is used for selecting a minimum absorbance value and a maximum absorbance value within the effective light absorption point range;
the ratio acquisition module is used for acquiring a difference value between the maximum absorbance value and the minimum absorbance value by taking the minimum absorbance value as a base number, and calculating the ratio of the difference value to the minimum absorbance value as the base number;
the reference value presetting module is used for presetting an agglutination reference value;
and the agglutination judgment module is used for comparing the ratio with the agglutination reference value, judging the blood in the sample hole to be agglutinated when the ratio is greater than the agglutination reference value, and judging the blood in the sample hole to be not agglutinated when the ratio is less than or equal to the agglutination reference value.
6. The apparatus according to claim 5, further comprising a reaction time setting module for setting a reaction time of a predetermined time period after the blood cells or serum of the known blood type are placed in the sample well and mixed with the blood cells or serum of the blood to be tested, and performing an absorbance scan on the mixed reaction sample after the reaction time is reached.
7. The apparatus according to claim 5, wherein the light absorption point scanning module scans the absorbance of the reaction sample in the sample well by a spectrophotometer.
8. The apparatus according to claim 7, wherein in the effective range defining module, the effective light absorption point range is a circular area having a radius r from the center point, the light absorption point is used as an effective light absorption point when the light absorption point is located within the circular area range, the light absorption point is used as an error light absorption point when the light absorption point is located outside the circular area range, and the light absorption value of the error light absorption point is rejected by the error point rejecting module.
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| WO2015190225A1 (en) * | 2014-06-12 | 2015-12-17 | コニカミノルタ株式会社 | Diagnosis-assistance-information generation method, image processing device, diagnosis-assistance-information generation system, and image processing program |
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| US4873633A (en) * | 1985-10-18 | 1989-10-10 | Cetus Corporation | User controlled off-center light absorbance reading adjuster in a liquid handling and reaction system |
| AU3057689A (en) * | 1988-02-03 | 1989-08-25 | Cetus Corporation | Apparatus and method for detecting hemagglutination reactions |
| US6330058B1 (en) * | 1999-04-14 | 2001-12-11 | University Of South Florida | Spectrophotometric method and apparatus for blood typing |
| JP4796265B2 (en) * | 2000-06-12 | 2011-10-19 | シスメックス株式会社 | Immunoassay method and immunoassay device |
| CN1448725A (en) * | 2002-04-01 | 2003-10-15 | 北京贝斯特医用仪器有限公司 | Method and apparatus for accomplishing ABO blood type full-automatic identification on full-automatic enzyme non-analysis meter |
| EP2085764A4 (en) * | 2006-11-15 | 2014-11-05 | Beckman Coulter Inc | Method for determining agglutination |
| CA2671078A1 (en) * | 2006-11-29 | 2008-06-05 | Julio Batistoni | Method for analyzing image data relating to agglutination assays |
| JP2011133364A (en) * | 2009-12-24 | 2011-07-07 | Beckman Coulter Inc | Method and apparatus for determiing hemagglutination image |
| CN107167616B (en) * | 2017-03-13 | 2019-06-07 | 中国科学院苏州生物医学工程技术研究所 | Blood type testing methods and device |
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| CN101346618A (en) * | 2005-12-23 | 2009-01-14 | 恩德莱斯和豪瑟尔测量及调节技术分析仪表两合公司 | Calibration in a laboratory reference method |
| WO2015190225A1 (en) * | 2014-06-12 | 2015-12-17 | コニカミノルタ株式会社 | Diagnosis-assistance-information generation method, image processing device, diagnosis-assistance-information generation system, and image processing program |
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