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A method suitable for rapid classification and counting of fish blood cells and its application

A rapid classification and blood cell technology, applied in the field of detection, to achieve the effect of good batch repeatability, easy access, and ensure accuracy and reliability

Active Publication Date: 2021-07-30
TIANJIN NORMAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0010] A method suitable for rapid classification and counting of fish blood cells, characterized in that: Aiming at the two main problems that the number and shape of nucleated red blood cells that cannot be removed interfere with the classification and counting of white blood cells, a method using fluorescent dyes and automatic image analysis is established. Combining measurement and analysis strategies, introducing DAPI dyes, using image-assisted automated analysis for the comparison of cell nucleus morphology and position differences, and further accurately identifying and delimiting the boundaries of cell groups through fusion images of bright field and fluorescence field of view

Method used

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  • A method suitable for rapid classification and counting of fish blood cells and its application
  • A method suitable for rapid classification and counting of fish blood cells and its application
  • A method suitable for rapid classification and counting of fish blood cells and its application

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Experimental program
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Effect test

Embodiment 1

[0032] Analysis of the health range of crucian carp hemogram

[0033] Similar to medical blood examination, the type and quantity of blood cells of each animal reflect the health status of its host to a certain extent, and the type and proportion range of various types of cells in the blood of healthy crucian carp can be obtained through scale measurement.

[0034] Detection process:

[0035] (1) Select 30 healthy crucian carp with a body weight of 300±30g, draw blood from the tail, draw 100 μL of whole blood, accurately absorb 20 μL of anticoagulated whole blood and fix it in 4% paraformaldehyde fixative solution for 5 minutes; centrifuge and discard Clear, resuspend the cell mass with DAPI staining solution, stain at room temperature for 5 minutes, and resuspend in PBS.

[0036] (2) Use a 70 μm cell sieve to filter out the adherent cells, and test on the machine; collect 30,000 cell samples at a medium rate.

[0037] (3) Analyze the data with IDEAS Application v6.1 softwar...

Embodiment 2

[0039] Hemogram Analysis of Carp Under Acute Ammonia Nitrogen Stress

[0040] The invention is not only applicable to crucian carp, but also applicable to the blood cell detection of carp and other fishes containing complete nuclei. Nitrogen-containing organic matter such as residual bait, feces, and fallen debris are all sources of ammonia nitrogen in water bodies. If the concentration of ammonia nitrogen exceeds a certain range, it will cause serious health threats to fish and other aquatic animals. These health damages are usually indicated by changes in blood patterns. sign.

[0041] (1) Set up a control group without ammonia nitrogen and an experimental group containing 200ppm ammonia nitrogen; select 10 healthy carps with a body weight of 500±50g, 5 for each, and place them in the water bodies of the control group and the experimental group respectively. After 3 days, test the blood Various changes in blood cell content

[0042] (2) Use the tail blood method to extract...

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Abstract

The invention discloses a high-throughput rapid classification and analysis method for blood cells suitable for common fish such as crucian carp and carp. Aiming at the problem that nucleated red blood cells that cannot be removed interfere with white blood cell classification and counting in terms of quantity and shape, an automated measurement and analysis strategy combining fluorescent dyes and multi-channel images is used to separate the interference The red blood cell samples are separated from the white blood cells, and the range of different white blood cell groups that undertake important immune functions such as lymphocytes, monocytes, and granulocytes is divided, and automatic analysis is completed. The difficulty of effectively removing nucleated red blood cells has long plagued the automatic analysis of blood images of many types of animals. This invention combines fluorescent dyes with image analysis, and uses the high-throughput and high-sensitivity measurement modules of the multi-dimensional panoramic analyzer to accurately identify red blood cell groups. Eliminating its interference provides important technical support for focused analysis of leukocyte groups and has broad application prospects.

Description

technical field [0001] The invention belongs to the technical field of detection, and relates to a method for analyzing the types, quantities and proportions of various blood cells in crucian carp whole blood by using a novel automatic cell detection technology combining fluorescent dyes and multi-channel images. The method is simple, fast, accurate, and high-throughput detection of crucian carp blood cell types and quantitative ratios, and can perform more accurate and efficient automatic analysis of crucian carp blood cells without effective cleavage reagents, avoiding heavy manual classification and counting, and Manual identification differences lead to "different" result deviations, which solves the main problem of interference with white blood cell classification and counting caused by a large number of red blood cells that cannot be removed, and realizes a rapid "blood test" for fish. Background technique [0002] Hemogram refers to the general term of the general tes...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/06
CPCG01N33/5094
Inventor 张亦陈张树花刘逸尘晋伟耿绪云孙金生
Owner TIANJIN NORMAL UNIVERSITY
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