虹鳟吞噬实验的优化

IF 0.6 4区农林科学 Q3 VETERINARY SCIENCES

Veterinarni Medicina Pub Date : 2021-06-01 DOI:10.17221/189/2020-VETMED

H. Minářová, P. Ondračkova, M. Palíková, J. Mareš, J. Blahova, K. Jarova, M. Faldyna

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引用次数: 0

摘要

吞噬试验是最常用的功能性免疫学方法之一。有许多可能的方法来评估白细胞及其摄取不同颗粒的能力。本研究的目的是利用标记酶酶体颗粒(Alexa Fluor 488和Texas Red偶联物)优化虹鳟(Oncorhynchus mykiss)的吞噬试验。全血与颗粒在不同条件下孵育，随后在低渗环境中溶血分离白细胞。采用流式细胞术观察不同孵育时间、温度、血容量和稀释度对细胞吞噬活性的影响。我们的实验表明，在15°C条件下孵育至少2 h的效果最佳，而血容量和稀释度没有显著影响。优化后的分析方法将用于鱼类健康检查和进一步的实验研究。

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Optimisation of phagocytosis assay in rainbow trout (Oncorhynchus mykiss)

A phagocytosis assay is one of the most commonly used functional immunological methods. There are many possible ways of assessing leucocytes and their ability to ingest different particles. The aim of this study was to optimise the phagocytosis assay in rainbow trout (Oncorhynchus mykiss) using labelled zymosan particles (Alexa Fluor 488 and Texas Red conjugate). Whole blood was incubated with the particles under different conditions and leucocytes were subsequently isolated by haemolysis in a hypotonic environment. The effect of the different incubation time, temperature, blood volume and dilution on the phagocytic activity was evaluated by flow cytometry. Our experiments showed that the incubation for at least 2 h at 15 °C provided optimal results, while the blood volume and dilution had no significant effect. The optimised assay will be used for the examination of fish health and in further experimental studies.

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来源期刊

Veterinarni Medicina Veterinary Sciences-兽医学

CiteScore

1.30

自引率

0.00%

发文量

审稿时长

18-36 weeks

期刊介绍： The journal Veterinarni Medicina publishes in English original papers, short communications, critical reviews and case reports from all fields of veterinary and biomedical sciences.