Method applicable for fish blood cell fast classification and counting and application thereof
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A rapid classification and blood cell technology, applied in the field of detection, to achieve the effect of less detection amount, strong competitive advantage and low cost
Active Publication Date: 2018-02-13
TIANJIN NORMAL UNIVERSITY
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[0011] 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 analys...
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Embodiment 1
[0033] Analysis on the health range of crucian carp blood picture
[0034] Similar to medical blood examination, the type and number of blood cells of each animal reflects the health of its host to a certain extent. The types and proportions of various types of cells in the blood of healthy crucian carp can be obtained through scale measurement.
[0035] Detection process:
[0036] (1) Select 30 healthy crucian carp weighing 300±30g, take blood from the tail, draw 100μL of whole blood, accurately draw 20μL of anticoagulated whole blood and fix it in 4% paraformaldehyde fixative for 5 minutes; centrifuge and discard. Clear, resuspend the cell pellet with DAPI staining solution, stain for 5 minutes at room temperature, and resuspend in PBS.
[0037] (2) Use a 70μm cell sieve to filter to remove adherent cells, and test on the machine; collect 30,000 cell samples at a medium rate.
[0038] (3) Analyze the data with IDEAS Application v6.1 software, circle the boundaries of specific cell gr...
Embodiment 2
[0040] Analysis of blood picture of carp under acute ammonia nitrogen stress
[0041] The invention is not only suitable for crucian carp, but also suitable for carp and other fish blood cell detection containing intact cell nuclei. Nitrogen-containing organic matter such as bait, feces, and shed debris are all sources of ammonia nitrogen in water bodies. Ammonia nitrogen concentration exceeding a certain range will cause serious health threats to fish and other aquatic animals. These health damages usually have obvious indicators in terms of blood changes. Levy.
[0042] (1) Set up a control group without ammonia nitrogen and an experimental group containing 200ppm ammonia nitrogen; select 10 healthy carps weighing 500±50g, take 5 from each, and place them in the water of the control group and the experimental group respectively, and test the blood after 3 days Various blood cell content changes
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Abstract
The invention discloses a method applicable for blood cell high-flux fast classification analysis of common fishes such as crucian and carps. By aiming at the problem of white blood cell classification and counting interference by removing-incapable nucleated red blood cells in two aspects of the quantity and the form, an automatic measurement and analysis strategy of combining the fluorochrome with the multi-channel images is utilized; the interference red blood cell sampling points are separated from the white blood cells according to the differences of particles in the cells, cell nucleus forms, positions and the like; different white blood cell class group range of lymphocyte, monocyte, particle cells and the like undertaking important immunologic functions is divided; the automatic analysis is completed. The difficulty that the nucleated red blood cells cannot be effectively removed troubles the automatic analysis of hemogram of various kinds of animals for a long time. The fluorochrome is combined with the imaging analysis; a high-flux and high-sensitivity measuring module of a multidimensional panoramic analyzer is used for accurately recognizing the red blood cell class groups; the interference is eliminated; the important technical support is provided for the focused analysis of the white blood cell class groups; wide application prospects are realized.
Description
[0001] This patent application was completed by Tianjin Normal University School of Life Sciences / Tianjin Key Laboratory of Animal and Plant Resistance, and received the National Natural Science Foundation of China (31472299), Tianjin Applied Basic and Frontier Technology Research Program (15JCZDJC33800), Tianjin Aquatic State and Aquaculture The key laboratory open fund (TJAE2015005), "Twelfth Five-Year" rural area national science and technology plan project (2011BAD13B04) funding. Technical field [0002] The invention belongs to the field of detection technology, and relates to a new type of automatic cell detection technology combining fluorescent dyes and multi-channel images, and a method for analyzing the types, numbers and proportions of various blood cells in the whole blood of crucian carp. This method can be simple, fast, accurate, and high-throughput detection of crucian carp blood cell types and number ratios, and can perform more accurate and efficient automated ana...
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