Method for differentiating shellfish granular blood cell types

Abstract

The invention relates to a method for differentiating a shellfish granular blood cell types. The method comprises the following steps of: (1) dissolving neutral red in a phosphate buffer solution at a mass ratio of 100:(1-3), filtering and then storing at a temperature of 4-5 DEG C, and diluting before use; (2) preparing a shellfish blood cell suspension, taking 1-2 ml of a neutral red working solution and 1-2 ml of the blood cell suspension and mixing evenly in a cell culture plate hole, dyeing for 30-40 minutes, then dropwise adding a drop of evenly mixed solution on a glass slide, and then observing the type of the shellfish granular blood cells through microscopic examination. Compared with the traditional MGG (May-Grunwald -Giemsa) dyeing method, the method provided by the invention is free of cell fixation, less in operation to the cells, and capable of clearly differentiating the granular blood cell type; therefore, a very convenient and practicable method is provided for studying the phagocytic capability, athletic capability, reactive oxygen generation capability and wound repair capability of eosinophilic granulocyte and basophilic granulocyte from the aspect of living cells.

Description

technical field [0001] The invention belongs to the field of shellfish hemocyte immunology, and particularly relates to a method for distinguishing shellfish granule blood cell types. Background technique [0002] Shellfish is the dominant species of mariculture in my country, and occupies a pivotal position in the mariculture industry. However, in recent years, with the blind expansion of aquaculture area, the continuous deterioration of the aquaculture ecological environment, and the lack of scientific and reasonable aquaculture planning and management, a series of common aquaculture problems have gradually emerged in my country's shellfish aquaculture industry. The shellfish germplasm degrades obviously, and the immunity and disease resistance of the young shellfish are reduced, and they are prone to disease. Aiming at this problem, there is no feasible control method at present, and a large part of the reason is that the immune and disease resistance mechanism of shellfi...

AI Technical Summary

Problems solved by technology

The currently commonly used cellular immunology methods often can only satisfy one of them: although the MGG method can accurately distinguish the two types of granulocytes, the cells stained by it are in a non-viable state after being fixed with ethanol; flow cytometry Although it can keep the cells under study alive, it cannot clearly distinguish the two types of granulocytes on the scatter plot

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