A method and application for rapid analysis of crustacean blood lymphocyte groups and quantity

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 ...

AI Technical Summary

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

However, this trial and error operation is often abandoned due to time-consuming and labor-intensive results and poor repeatability of results.

Due to the limitations of the above-mentioned factors, the research on the analysis of crustacean blood lymphocytes by conventional flow cytometry has been in the experimental stage, and there have been no general methods and commercial auxiliary detection products reported so far.

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