CN113552126A - Reticulocyte detection method and system - Google Patents

Reticulocyte detection method and system Download PDF

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CN113552126A
CN113552126A CN202110836014.7A CN202110836014A CN113552126A CN 113552126 A CN113552126 A CN 113552126A CN 202110836014 A CN202110836014 A CN 202110836014A CN 113552126 A CN113552126 A CN 113552126A
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image
sample
detected
reticulocyte
reticulocytes
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黄来荣
王娅
雷礼年
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Fuzhou Jinyu Medical Laboratory Co ltd
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Fuzhou Jinyu Medical Laboratory Co ltd
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    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/84Systems specially adapted for particular applications
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00Sampling; Preparing specimens for investigation
    • G01N1/28Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N1/2813Producing thin layers of samples on a substrate, e.g. smearing, spinning-on
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00Sampling; Preparing specimens for investigation
    • G01N1/28Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N1/30Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
    • GPHYSICS
    • G06COMPUTING; CALCULATING OR COUNTING
    • G06TIMAGE DATA PROCESSING OR GENERATION, IN GENERAL
    • G06T7/00Image analysis
    • G06T7/0002Inspection of images, e.g. flaw detection
    • G06T7/0012Biomedical image inspection
    • GPHYSICS
    • G06COMPUTING; CALCULATING OR COUNTING
    • G06TIMAGE DATA PROCESSING OR GENERATION, IN GENERAL
    • G06T7/00Image analysis
    • G06T7/10Segmentation; Edge detection
    • G06T7/11Region-based segmentation
    • GPHYSICS
    • G06COMPUTING; CALCULATING OR COUNTING
    • G06TIMAGE DATA PROCESSING OR GENERATION, IN GENERAL
    • G06T7/00Image analysis
    • G06T7/10Segmentation; Edge detection
    • G06T7/155Segmentation; Edge detection involving morphological operators
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00Sampling; Preparing specimens for investigation
    • G01N1/28Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N1/30Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
    • G01N2001/302Stain compositions
    • GPHYSICS
    • G06COMPUTING; CALCULATING OR COUNTING
    • G06TIMAGE DATA PROCESSING OR GENERATION, IN GENERAL
    • G06T2207/00Indexing scheme for image analysis or image enhancement
    • G06T2207/30Subject of image; Context of image processing
    • G06T2207/30242Counting objects in image

Abstract

The invention relates to a reticulocyte detection method and a reticulocyte detection system, wherein the method comprises the following steps: collecting a sample to be detected, labeling the sample to be detected, and dyeing a sample slide by adopting a reticulocyte dyeing agent; manufacturing a dyed sample slide by using the dyed sample to be detected; performing laser scattering on the dyed sample slide, performing image amplification on the dyed sample slide by using a microscope, and performing image acquisition by using digital equipment; processing the acquired images, and performing statistics to obtain the number of reticulocytes and the number of red blood cells; measuring the hemoglobin concentration of a sample to be detected; calculating parameters of the reticulocytes according to the obtained number of the reticulocytes, the obtained number of the erythrocytes and the hemoglobin concentration; according to the invention, the reticulocyte stain is adopted to stain the sample to be detected, so that the influence of fluorescence generated by red blood cells on the detection result is avoided, and the error is small.

Description

Reticulocyte detection method and system
Technical Field
The invention relates to the technical field of medical detection, in particular to a reticulocyte detection method and a reticulocyte detection system.
Background
Reticulocytes are not fully mature red blood cells, and the value in the peripheral blood can reflect the generating function of bone marrow erythrocytes, so that the reticulocytes have important significance for the diagnosis of hematopathy and the observation of treatment response, and can reflect the hematopoietic function of bone marrow erythropoiesis and judge the curative effect of anemia and related diseases.
In the existing reticulocyte detection technology, the reticulocyte is usually detected by adopting flow cytometry, and the reticulocyte detection technology is easily influenced by the fluorescence generated by the erythrocyte and has larger error.
Disclosure of Invention
In order to overcome the technical defects in the prior art, the invention provides a reticulocyte detection method and a reticulocyte detection system, which can effectively solve the problems in the background art.
In order to solve the technical problems, the technical scheme provided by the invention is as follows:
the embodiment of the invention discloses a reticulocyte detection method and a reticulocyte detection system.
Preferably in any of the above aspects, the method comprises the steps of:
the method comprises the following steps: collecting a sample to be detected, labeling the sample to be detected, and dyeing a sample slide by adopting a reticulocyte dyeing agent;
step two: manufacturing a dyed sample slide by using the dyed sample to be detected;
step three: performing laser scattering on the dyed sample slide, performing image amplification on the dyed sample slide by using a microscope, and performing image acquisition by using digital equipment;
step four: processing the acquired images, and performing statistics to obtain the number of reticulocytes and the number of red blood cells;
step five: measuring the hemoglobin concentration of a sample to be detected;
step six: and calculating the related parameters of the reticulocytes according to the obtained number of the reticulocytes, the number of the erythrocytes and the hemoglobin concentration.
In any of the above embodiments, it is preferred that the reticulocyte stain is T03-3392-5O reticulocyte stain, and the T03-3392-5O reticulocyte stain consists of N-tetradecyl-N, N dimethyl-3 amino-1 propionic acid inner spirit and oxa howl 750(Oxazine750) dye.
In any of the above embodiments, it is preferred that in the T03-3392-5O reticulocyte stain, the N-tetradecyl-N and N-dimethyl-3 amino-1 propionic acid inner spirit is used to turn the erythrocytes into globotrys, and the oxazole howl 750 is a nucleic acid agent used to selectively stain reticulocytes.
In any of the above embodiments, preferably, the staining of the sample slide with the reticulocyte stain comprises the following steps: mixing a T03-3392-5O reticulocyte stain and a sample to be detected, wherein the ratio of the T03-3392-5O reticulocyte stain to the sample to be detected is 1:1, and standing for ten minutes to fifteen minutes at room temperature to obtain the sample to be detected after staining.
In any of the above-described embodiments, it is preferable that, when the sample slide is stained with the reticulocyte stain, the staining standing time is increased when the room temperature is low, and the staining standing time is decreased when the room temperature is high.
In any of the above protocols, it is preferred that in making a sample smear, the method comprises the steps of: placing a drop of blood in a sample to be detected at one centimeter of one end of a glass slide, horizontally holding the glass slide by the left hand, holding a picture by the right hand from the back to approach the blood drop, expanding the blood to a certain width along the edge of a push sheet, immediately inclining the push sheet to ensure that the push sheet and the glass slide support to form an angle of thirty to forty-five degrees, and slightly pressing the edge of the push sheet to push the blood to be a sample smear with proper thickness.
In any of the above schemes, preferably, the method includes the following steps when processing the acquired image:
the method comprises the following steps: preprocessing the acquired image;
step two: carrying out image form transformation on the preprocessed image;
step three: and performing image feature extraction on the image subjected to image form transformation.
In any of the above schemes, preferably, when the acquired image is preprocessed, the method includes the following steps:
the method comprises the following steps: carrying out denoising processing on the acquired image by using a median filtering method, wherein a definition formula of the median filtering method is as follows: r ═ med { a ═k∣k=1,2,3,4,……,n};
Step two: carrying out graying processing on the obtained video image;
step three: performing mathematical morphology binarization processing on the video image subjected to the graying processing;
step four: and performing thresholding operation on the image after the graying processing and the binarization processing of mathematical morphology, and converting the image into a binary image.
In any of the above embodiments, preferably, when the thresholding operation is performed, two different sets of threshold values are selected, and the thresholding operation is performed twice on the image to obtain a binary image containing the red blood cell image and a binary image containing no red blood cell image, respectively.
In any of the above embodiments, it is preferable that the processing is performed by a cell segmentation method based on a pit point when the acquired image is subjected to image morphological transformation.
In any of the above aspects, preferably, when the image feature extraction is performed on the preprocessed image, the extracted features include reticulocyte contours and red blood cell contours.
In any of the above schemes, preferably, when performing statistics, the method includes the following steps:
the method comprises the following steps: setting the gray value of a target area of the image as 0 and the gray value of a background area as 1;
step two: scanning the binarized image from left to right and from top to bottom, if the gray value of the current pixel point is 1, scanning the next pixel, otherwise, checking the gray value of the adjacent pixel at the left, upper right and upper right, storing, if the gray values of the pixel at the left, upper right and upper right are all 1, then adding a mark, if the gray value of only one pixel point A in the adjacent pixel points is 0, the marking value of the pixel point A is given to the current pixel point, if the gray value of the left, upper right pixel points of the current pixel point is more than 4 and B >1 pixel points is 0, and according to the priority principle of the scanning sequence of left, upper right and upper right, the marking values of the left, upper right and upper right pixel points of the current pixel point are given to the current pixel point, the equivalent pair is made on the marking value of the pixel point, and the marking values are put into an equivalent array.
Step three: and carrying out secondary scanning, sorting out an equivalence relation according to the equivalence pair array, and then re-marking the target area according to the equivalence relation.
Step four: and (4) performing statistical integration on the markers in the image to respectively obtain the number n of reticulocytes and the total number m of erythrocytes.
In any of the above schemes, it is preferable that when the hemoglobin concentration of the sample to be detected is measured, the method includes the following steps: selecting a sample to be detected, adding hemolysin and the sample to be detected into a colorimetric pool, penetrating the colorimetric pool by using monochromatic light, and calculating the concentration of hemoglobin according to a formula.
In any of the above solutions, it is preferable that the expression is given by
Figure BDA0003177092090000041
Calculating the concentration of hemoglobin in a sample to be detected; wherein HGB is the concentration of hemoglobin in a sample to be detected, and K is an absorption constant; l isinIs the input light intensity; l isoutTo output light intensity.
In any of the above schemes, preferably, after the obtained image is subjected to thresholding operation, the obtained binary image is counted, and the number of pixel points with gray levels greater than the threshold is counted to obtain the total area occupied by the reticulocytes in the image.
In any of the above embodiments, preferably, the parameters related to reticulocytes include reticulocyte mean cell volume, cell mean hemoglobin concentration, cell hemoglobin content, reticulocyte absolute value, red blood cell absolute value, and reticulocyte concentration.
In any of the above aspects, it is preferred that the reticulocyte detection system comprises:
the collection module is used for collecting a blood sample of a person to be detected;
the dyeing module is used for dyeing the sample to be detected collected by the collecting module;
the image amplification and acquisition module is used for carrying out image acquisition and amplification on the dyed sample to be detected;
the image processing module is used for processing the image acquired by the image amplifying and acquiring module;
the calculation module is used for calculating the parameters of the reticulocytes according to the image processed by the image processing module;
the detection module is used for calculating the concentration of hemoglobin in a sample to be detected;
and the statistical module is used for counting and integrating the data obtained by the calculation module and the detection module.
In any of the above aspects, it is preferable that the image magnifying and acquiring means includes a microscope and a color camera.
In any of the above schemes, it is preferable that the apparatus further includes a storage module and a display module, the storage module is configured to store parameters of the sample to be detected, and the display module is configured to enable a user to read data stored in the storage module.
Compared with the prior art, the invention has the beneficial effects that:
1. the method adopts the reticulocyte stain to stain the sample to be detected, so that the red blood cells can be expanded into a spherical shape, and the influence of the shape of the red blood cells on the detection result is avoided.
2. According to the invention, the reticulocyte stain is adopted to stain the sample to be detected, so that the influence of fluorescence generated by red blood cells on the detection result is avoided, and the error is small.
3. The invention adopts a secondary scanning method to carry out identification statistics on the number of the reticulocytes, and has high accuracy and high speed.
Drawings
The drawings are included to provide a further understanding of the invention, and are incorporated in and constitute a part of this specification.
FIG. 1 is a schematic diagram of a method for detecting reticulocytes according to an embodiment of the present invention;
FIG. 2 is a flowchart illustrating the reticulocyte counting process in a reticulocyte detection method according to an embodiment of the present invention;
FIG. 3 is a schematic diagram of a reticulocyte detection system according to an embodiment of the present invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is described in further detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
It will be understood that when an element is referred to as being "secured to" or "disposed on" another element, it can be directly on the other element or be indirectly on the other element. When an element is referred to as being "connected to" another element, it can be directly connected to the other element or be indirectly connected to the other element.
In the description of the present invention, it is to be understood that the terms "length", "width", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", and the like, indicate orientations or positional relationships based on the orientations or positional relationships illustrated in the drawings, and are used merely for convenience in describing the present invention and for simplicity in description, and do not indicate or imply that the devices or elements referred to must have a particular orientation, be constructed in a particular orientation, and be operated, and thus, are not to be construed as limiting the present invention.
Furthermore, the terms "first", "second" and "first" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include one or more of that feature. In the description of the present invention, "a plurality" means two or more unless specifically defined otherwise.
For better understanding of the above technical solutions, the technical solutions of the present invention will be described in detail below with reference to the drawings and the detailed description of the present invention.
A reticulocyte detection method and a reticulocyte detection system thereof are provided, wherein the method comprises the following steps:
the method comprises the following steps: collecting a sample to be detected, labeling the sample to be detected, and dyeing a sample slide by adopting a reticulocyte dyeing agent;
step two: manufacturing a dyed sample slide by using the dyed sample to be detected;
step three: performing laser scattering on the dyed sample slide, performing image amplification on the dyed sample slide by using a microscope, and performing image acquisition by using digital equipment;
step four: processing the acquired images, and performing statistics to obtain the number of reticulocytes and the number of red blood cells;
step five: measuring the hemoglobin concentration of a sample to be detected;
step six: and calculating the related parameters of the reticulocytes according to the obtained number of the reticulocytes, the number of the erythrocytes and the hemoglobin concentration.
The principle of the method is to identify reticulocytes and mature red blood cells by increasing light absorption, the light absorption quantity is in proportional relation with the quantity of RNA existing in cells, and the method detects the light absorption quantity of RNA and reticulocyte reagent deposited in the cells, but not fluorescence, so that the method overcomes the error of a flow cytometer caused by the fluorescence generated by the red blood cells in some cases, and can count the percentage and absolute value of the reticulocytes in 2 ten thousand red blood cells in a few seconds.
Specifically, the reticulocyte stain is T03-3392-5O reticulocyte stain, and the T03-3392-5O reticulocyte stain consists of N-tetradecyl-N, N dimethyl-3 ammonia-1 propionic acid inner spirit and oxadiazine 750 dye.
In the T03-3392-5O reticulocyte stain, the N-tetradecyl-N and N-dimethyl-3 amino-1 propionic acid internal spirit is used for changing erythrocytes into Tong, and the oxazole howl 750 is a nucleic acid reagent used for selectively staining reticulocytes.
Specifically, when the sample slide is stained by using the reticulocyte stain, the method comprises the following steps: mixing a T03-3392-5O reticulocyte stain and a sample to be detected, wherein the ratio of the T03-3392-5O reticulocyte stain to the sample to be detected is 1:1, and standing for ten minutes to fifteen minutes at room temperature to obtain the sample to be detected after staining.
Further, when the sample slide is stained with the reticulocyte stain, the staining standing time is increased when the indoor temperature is low, and the staining standing time is decreased when the indoor temperature is high.
Specifically, when the sample smear is made, the method comprises the following steps: placing a drop of blood in a sample to be detected at one centimeter of one end of a glass slide, horizontally holding the glass slide by the left hand, holding a picture by the right hand from the back to approach the blood drop, expanding the blood to a certain width along the edge of a push sheet, immediately inclining the push sheet to ensure that the push sheet and the glass slide support to form an angle of thirty to forty-five degrees, and slightly pressing the edge of the push sheet to push the blood to be a sample smear with proper thickness.
The shape of the red blood cells is changed to spherical red blood cells, elliptical red blood cells, target red blood cells, lip red blood cells, sickle red blood cells, echinocyte, schizored blood cells, rouleaux red blood cells and nucleated red blood cells, and the shapes of all the red blood cells in the images are circular when the images of the sample smears are processed because the red blood cells are changed to spherical shapes by using the spirit of N-tetradecyl-N and N-dimethyl-3 amino-1 propionic acid, so that the shapes of the red blood cells in the images are not required to be added into the calculation process.
Specifically, the digital device is a color camera.
Specifically, when processing the acquired image, the method comprises the following steps:
the method comprises the following steps: preprocessing the acquired image;
step two: carrying out image form transformation on the preprocessed image;
step three: and performing image feature extraction on the image subjected to image form transformation.
The image preprocessing relates to image processing modes such as denoising, graying, binarization and the like of an image, and aims to enable the image to be convenient for recognition of cells, after the image form transformation processing, the cells are transformed into independent recognition entities, so that independent cell individuals are further segmented from the image, characteristics such as area, outline, color and the like of the cells are extracted, the image is convenient to operate in the next step, and the collected original image has the problems of adhesion, blurring and the like due to the large number and density of the cells, so that the image form transformation is needed before recognition.
Specifically, when the acquired image is preprocessed, the method comprises the following steps:
the method comprises the following steps: carrying out denoising processing on the acquired image by using a median filtering method, wherein a definition formula of the median filtering method is as follows: r ═ med { a ═k∣k=1,2,3,4,……,n};
Step two: carrying out graying processing on the obtained video image;
step three: performing mathematical morphology binarization processing on the video image subjected to the graying processing;
step four: and performing thresholding operation on the image after the graying processing and the binarization processing of mathematical morphology, and converting the image into a binary image.
Further, when the threshold operation is carried out, two groups of different threshold values are selected, and the threshold operation is carried out on the image twice, so that a binary image containing the red blood cell image and a binary image containing no red blood cell image are respectively obtained.
Specifically, when the acquired image is subjected to image morphological transformation, a cell segmentation method based on concave points is adopted for processing.
Specifically, when image feature extraction is performed on the image after the preprocessing, the extracted features include a reticulocyte contour and a red blood cell contour.
Specifically, when performing statistics, the method comprises the following steps:
the method comprises the following steps: setting the gray value of a target area of the image as 0 and the gray value of a background area as 1;
step two: scanning the binarized image from left to right and from top to bottom, scanning the next pixel if the gray value of the current pixel is 1, otherwise, checking the gray value of the adjacent pixel at the left, upper right and upper right, storing, adding a mark if the gray values of the pixels at the left, upper right and upper right are all 1, if the gray value of only one pixel A in the adjacent pixels is 0, giving the mark value of the pixel A to the current pixel, if the pixels at the left, upper right and upper right of the current pixel have the gray values of B pixels of 0, wherein 4> B >1, giving the mark values of the pixels at the left, upper right and upper left of the current pixel to the current pixel according to the priority of the scanning sequence left, upper right and upper right, and making an equivalence pair for the mark value of the pixel, it is put into an equivalent array.
Step three: and carrying out secondary scanning, sorting out an equivalence relation according to the equivalence pair array, and then re-marking the target area according to the equivalence relation.
Step four: and (4) performing statistical integration on the markers in the image to respectively obtain the number n of reticulocytes and the total number m of erythrocytes.
Specifically, when the hemoglobin concentration of a sample to be detected is measured, the method comprises the following steps: selecting a sample to be detected, adding hemolysin and the sample to be detected into a colorimetric pool, penetrating the colorimetric pool by using monochromatic light, and calculating the concentration of hemoglobin according to a formula.
In the process of measuring hemoglobin, firstly, hemolysin is added into a blood sample to break cell membranes of red blood cells and release the hemoglobin; then, the hemolysin reacts with hemoglobin to generate cyanided hemoglobin; and finally, monochromatic light with certain intensity penetrates through the colorimetric pool, and the input illumination intensity and the output illumination intensity are compared to finish the measurement of the concentration of the hemoglobin.
Further, by the formula
Figure BDA0003177092090000111
Calculating the concentration of hemoglobin in a sample to be detected; wherein HGB is the concentration of hemoglobin in a sample to be detected, and K is an absorption constant;
Linis the input light intensity; l isoutTo output light intensity.
Specifically, after thresholding operation is performed on the obtained image, the obtained binary image is counted, and the number of pixel points with gray levels larger than a threshold value is counted to obtain the total area occupied by the reticulocyte in the image.
Specifically, the parameters related to the reticulocytes include reticulocyte mean cell volume, cell mean hemoglobin concentration, cell hemoglobin content, reticulocyte absolute value, erythrocyte absolute value and reticulocyte concentration.
A reticulocyte detection system, the system comprising:
the collection module is used for collecting a blood sample of a person to be detected;
the dyeing module is used for dyeing the sample to be detected collected by the collecting module;
the image amplification and acquisition module is used for carrying out image acquisition and amplification on the dyed sample to be detected;
the image processing module is used for processing the image acquired by the image amplifying and acquiring module;
the calculation module is used for calculating the parameters of the reticulocytes according to the image processed by the image processing module;
the detection module is used for calculating the concentration of hemoglobin in a sample to be detected;
and the statistical module is used for counting and integrating the data obtained by the calculation module and the detection module.
Further, the image amplifying and collecting module comprises a microscope and a color camera.
The device further comprises a storage module and a display module, wherein the storage module is used for storing the parameters of the sample to be detected, and the display module is used for reading the data stored in the storage module by a user.
Compared with the prior art, the invention has the beneficial effects that:
1. the method adopts the reticulocyte stain to stain the sample to be detected, so that the red blood cells can be expanded into a spherical shape, and the influence of the shape of the red blood cells on the detection result is avoided.
2. According to the invention, the reticulocyte stain is adopted to stain the sample to be detected, so that the influence of fluorescence generated by red blood cells on the detection result is avoided, and the error is small.
3. The invention adopts a secondary scanning method to carry out identification statistics on the number of the reticulocytes, and has high accuracy and high speed.
Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art will understand that various changes, modifications and substitutions can be made without departing from the spirit and scope of the invention as defined by the appended claims. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims (10)

1. A reticulocyte detection method is characterized in that: the method comprises the following steps:
the method comprises the following steps: collecting a sample to be detected, labeling the sample to be detected, and dyeing a sample slide by adopting a reticulocyte dyeing agent;
step two: manufacturing a dyed sample slide by using the dyed sample to be detected;
step three: performing laser scattering on the dyed sample slide, performing image amplification on the dyed sample slide by using a microscope, and performing image acquisition by using digital equipment;
step four: processing the acquired images, and performing statistics to obtain the number of reticulocytes and the number of red blood cells;
step five: measuring the hemoglobin concentration of a sample to be detected;
step six: and calculating parameters of the reticulocytes according to the obtained number of the reticulocytes, the number of the erythrocytes and the hemoglobin concentration.
2. The method for detecting reticulocytes of claim 1, wherein: the reticulocyte stain is T03-3392-5O reticulocyte stain, and the T03-3392-5O reticulocyte stain consists of N-tetradecyl-N, N dimethyl-3 ammonia-1 propionic acid internal spirit and oxazole howl 750 dye.
3. The method for detecting reticulocytes of claim 2, wherein: when the sample slide is stained by adopting the reticulocyte stain, the method comprises the following steps: mixing a T03-3392-5O reticulocyte stain and a sample to be detected, wherein the ratio of the T03-3392-5O reticulocyte stain to the sample to be detected is 1:1, and standing for ten minutes to fifteen minutes at room temperature to obtain the sample to be detected after staining.
4. The method for detecting reticulocytes of claim 3, wherein: when making a sample smear, the method comprises the following steps: placing a drop of blood in a sample to be detected at one centimeter of one end of a glass slide, horizontally holding the glass slide by the left hand, holding a picture by the right hand from the back to approach the blood drop, expanding the blood to a certain width along the edge of a push sheet, immediately inclining the push sheet to ensure that the push sheet and the glass slide support to form an angle of thirty to forty-five degrees, and slightly pressing the edge of the push sheet to push the blood to be a sample smear with proper thickness.
5. The method for detecting reticulocytes of claim 4, wherein: when the acquired image is processed, the method comprises the following steps:
the method comprises the following steps: preprocessing the acquired image;
step two: carrying out image form transformation on the preprocessed image;
step three: and performing image feature extraction on the image subjected to image form transformation.
6. The method for detecting reticulocytes of claim 5, wherein: when the acquired image is preprocessed, the method comprises the following steps:
the method comprises the following steps: carrying out denoising processing on the acquired image by using a median filtering method, wherein a definition formula of the median filtering method is as follows: r ═ med { a ═k∣k=1,2,3,4,……,n};
Step two: carrying out graying processing on the obtained video image;
step three: performing mathematical morphology binarization processing on the video image subjected to the graying processing;
step four: and performing thresholding operation on the image after the graying processing and the binarization processing of mathematical morphology, and converting the image into a binary image.
7. The method for detecting reticulocytes of claim 6, wherein: when the threshold operation is carried out, two groups of different threshold values are selected, and the threshold operation is carried out on the image twice, so that a binary image containing the red blood cell image and a binary image containing no red blood cell image are respectively obtained.
8. The method for detecting reticulocytes of claim 7, wherein:
the method comprises the following steps: setting the gray value of a target area of the image as 0 and the gray value of a background area as 1;
step two: scanning the binarized image from left to right and from top to bottom, if the gray value of the current pixel point is 1, scanning the next pixel point, otherwise, checking the gray value of the adjacent pixel points of the left, the upper right and the upper right, storing, if the gray values of the pixel points of the left, the upper right and the upper right are all 1, adding a mark, if the gray value of only one pixel point A in the adjacent pixel points is 0, giving the mark value of the pixel point A to the current pixel point, and if the gray value of B pixel points in the pixel points of the left, the upper right and the upper right of the current pixel point is 0; wherein, if B is more than 1 and less than 5, the marking values of the left, upper right and upper right pixel points of the current pixel point are given to the current pixel point according to the priority principle of the scanning sequence of left, upper right and upper right, and the marking value of the pixel point is subjected to equivalence pair and is put into an equivalence array.
Step three: and carrying out secondary scanning, sorting out an equivalence relation according to the equivalence pair array, and then re-marking the target area according to the equivalence relation.
Step four: and (4) performing statistical integration on the markers in the image to respectively obtain the number n of reticulocytes and the total number m of erythrocytes.
9. The method for detecting reticulocytes of claim 8, wherein: when the hemoglobin concentration of a sample to be detected is measured, the method comprises the following steps: selecting a sample to be detected, adding hemolysin and the sample to be detected into a colorimetric pool, penetrating the colorimetric pool with monochromatic light, calculating hemoglobin concentration according to a formula, and calculating the hemoglobin concentration according to the formula
Figure FDA0003177092080000031
Calculating the concentration of hemoglobin in a sample to be detected; wherein HGB is the concentration of hemoglobin in a sample to be detected, and K is an absorption constant; l isinIs the input light intensity; l isoutTo output light intensity.
10. A reticulocyte detection system is characterized in that: the system comprises:
the collection module is used for collecting a blood sample of a person to be detected;
the dyeing module is used for dyeing the sample to be detected collected by the collecting module;
the image amplification and acquisition module is used for carrying out image acquisition and amplification on the dyed sample to be detected;
the image processing module is used for processing the image acquired by the image amplifying and acquiring module;
the calculation module is used for calculating the parameters of the reticulocytes according to the image processed by the image processing module;
the detection module is used for calculating the concentration of hemoglobin in a sample to be detected;
and the statistical module is used for counting and integrating the data obtained by the calculation module and the detection module.
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