Method of quickly analyzing group and number of blood lymphocytes of crustaceans and application of the method

A technology for blood lymphocytes and crustaceans, applied in the field of detection, can solve the problems of poor repeatability of results, lack of image reference, time-consuming and labor-intensive, etc., and achieve the effects of low cost, simplified operation process, and low dosage.

Active Publication Date: 2017-12-19
TIANJIN NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to the lack of image references, screening objects without special markers can only be detected repeatedly through the process of sample loading-recovery-analysis
In this process, cells cannot be reused, and due to the lack of ability to recover and analyze individual cells, the number of border cells and low-abundance cells is often small, so a large number of samples are required to try again and again
How

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  • Method of quickly analyzing group and number of blood lymphocytes of crustaceans and application of the method
  • Method of quickly analyzing group and number of blood lymphocytes of crustaceans and application of the method
  • Method of quickly analyzing group and number of blood lymphocytes of crustaceans and application of the method

Examples

Experimental program
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Effect test

Embodiment 1

[0045] Analysis of Hemolymphocyte Response to Acute Blood Loss Stimulation of Litopenaeus vannamei

[0046] (1) Experimental design

[0047] This method takes the granular structure in the cells of crustaceans such as shrimp and crabs as a reference for the classification of hemolymphocytes, so it is also suitable for the classification and analysis of hemolymphocytes of Litopenaeus vannamei containing particles in the cells. Ten healthy Litopenaeus vannamei with a body weight of 25±3g were randomly selected, among which 5 were the control group and 5 were the experimental group. At the beginning of the experiment, hemolymph was extracted from each animal in the experimental group according to 3% of body weight (v / w) to establish an acute massive blood loss model. After 12 hours, hemolymph samples were taken from each animal in the control group and the experimental group, and the proportion changes of various types of hemolymphocytes were detected according to the method of ...

Embodiment 2

[0055] Analysis of Hemolymph Response to White Spot Baculovirus (WSSV) Infection in Procambarus clarkii

[0056] White spot baculovirus is the main pathogen that harms prawn farming, and has brought huge losses to prawn farming for a long time. Procambarus clarkii is also the host of this pathogen. Therefore, through quantitative infection analysis, we can know the hemolymphocytes that the virus mainly infects. .

[0057] (1) Experimental design

[0058] Randomly select 10 healthy Procambarus clarkii with a body weight of 20±2g, set up a control group and an experimental group injected with WSSV virus, with 5 experimental animals in each group, take samples after 1 week, and analyze the changes in the ratio of the four types of hemolymph cells .

[0059] (2) Sample preparation

[0060] Carefully extract about 100 microliters of hemolymph from each animal in the control group and experimental group with a syringe containing anticoagulant, quickly transfer it into the fixativ...

Embodiment 3

[0066] Differentiation and Analysis of Blood Lymphocytes in Eriocheir sinensis

[0067] The normal range can be obtained by randomly measuring the proportion of each group of blood lymphocytes in healthy mitten crabs in batches, and it can be used for breeding monitoring and evaluation.

[0068] (1) Experimental design

[0069] Randomly select 10 Chinese mitten crabs weighing 20±2g, with complete appearance and good vitality.

[0070] (2) Carefully extract about 100 microliters of hemolymph from each animal in the control group and the experimental group with a syringe containing anticoagulant, quickly transfer it into the fixative, mix well, and fix at room temperature for 5 minutes

[0071] (3) Sample preparation

[0072] Carefully extract about 100 microliters of hemolymph from each animal in the control group and experimental group with a syringe containing anticoagulant, quickly transfer it into the fixative solution, mix well, and fix at room temperature for 5 minutes ...

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Abstract

The invention discloses a method of quickly analyzing group and number of blood lymphocytes of various common crustaceans according to the principles that with the particle structure in the blood lymphocytes of crustaceans, such as litopenaeus vannamei, procambarus clarkia, eriocheir sinensis and the like, as classification reference, the blood lymphocytes, in different groups, can be discriminated and statistic analysis is automatically completed at the same time with a region, where sample points are located, and auxiliary images as evidences by means of a multi-dimensional panorama analyzer having an image auxiliary function. The method is free of dyeing and avoids influence due to manual steps such as normal dyeing, observation under a microscope and the like; by means of automatic signal analysis of a high-sensitivity detector, the method has good repeatability. The multi-dimensional panorama analyzer has high-throughput detection performance, so that the reliability of a result from tens of thousands cell sample points is far higher than that of manual counting. The method is very low in usage amount of reagents and consumables and is low in cost due to trace amount detection. The method not only can be used for scientific analysis and daily monitoring, but also can supply technical support to classifying collection of the blood lymphocytes, so that the method has great application value.

Description

[0001] This patent application was completed by the School of Life Sciences of Tianjin Normal University / Tianjin Key Laboratory of Animal and Plant Resistance, and was supported by the National Natural Science Foundation of China (31472299), Tianjin Applied Basic and Frontier Technology Research Program (15JCZDJC33800), Tianjin Aquatic Ecology and Breeding Funding from the Key Laboratory Open Fund (TJAE2015005), and the "Twelfth Five-Year" National Science and Technology Program in Rural Areas (2011BAD13B04). technical field [0002] The invention belongs to the technical field of detection, based on the characteristics of intracellular particles, combined with the image-assisted analysis function of a multi-dimensional panoramic detector, to accurately detect the hemolymphocytes of common crustaceans such as Litopenaeus vannamei, Procambarus clarkii and Chinese mitten crab. Classification and ratio analysis, to establish a high-efficiency blood image detection method suitable ...

Claims

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Application Information

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IPC IPC(8): G01N33/483G01N21/84
CPCG01N21/84G01N33/4833
Inventor 张亦陈胡锦丽刘逸尘晋伟耿绪云孙金生
Owner TIANJIN NORMAL UNIVERSITY
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