Flow cytometry is an effective and widely used tool for determination of ploidy in fish, but it is not always possible to access the fresh samples for analysis. We investigated the potential for extended storage of fish tissue with sterlet and tench as representative species of Chondrostei and Teleostei, using blood and fin of subadult/adult specimens and tail of larvae. Thirteen procedures for extending storage, selected for rapidity and simplicity in both field and laboratory conditions, were tested for each tissue sample. Flow cytometry was applied to fresh tissue immediately after sampling and to tissue subjected to experimental protocols, always along with species-specific standard, after 1, 5, and 10 days storage at 0–4°C or freezing at −80°C. The fluorochrome 4′,6-diamidine-2′-phenylindole dihydrochloride was used with excitation/emission maximum 358/461 nm. Based on the measurability of stored samples, evaluation of directly measured coefficients of variation of their DNA peaks and the changes in fluorescence intensity compared to fresh tissue, optimal procedures for extended storage of the selected tissue types of the model species are suggested. © 2020 The Authors. Cytometry Part A published by Wiley Periodicals LLC. on behalf of International Society for Advancement of Cytometry.

中文翻译：

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通过流式细胞术测量 DNA 含量的鱼类样品的简单现场储存

流式细胞术是确定鱼类倍性的有效且广泛使用的工具，但并不总是能够获得新鲜样品进行分析。我们使用亚成体/成体标本的血液和鳍以及幼虫的尾巴，研究了以小背鱼和丁丁鱼为代表的软骨鱼和硬骨鱼的鱼类组织的长期储存潜力。每个组织样本均测试了 13 种延长储存时间的程序，这些程序是为了在现场和实验室条件下快速且简单地选择的。在采样后立即对新鲜组织和经过实验方案的组织进行流式细胞术，始终与物种特异性标准一起，在 0–4°C 储存 1、5 和 10 天或在 -80°C 冷冻后进行。使用荧光染料 4',6-二脒-2'-苯基吲哚二盐酸盐，激发/发射最大值为 358/461 nm。基于存储样本的可测量性、直接测量的 DNA 峰值变异系数的评估以及与新鲜组织相比荧光强度的变化，建议了模型物种所选组织类型的长期存储的最佳程序。© 2020 作者。细胞计数 A 部分由 Wiley periodicals LLC 出版。代表国际细胞计数促进会。