标题：Sysmex Partec 倍性分析仪在鱼类样本DNA含量测量中的应用
Abstract
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℃ or freezing at −80℃. 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.
Keywords: fixation; preservation; sterlet; tench; blood; fin tissue; larva tail tissue; coefficient of
variation; fluorescence intensity
摘要：
倍体分析仪是一种有效且广泛使用的鱼类倍性测定工具，但并不总是能够获取新鲜样本进行分析。我们使用亚成体/成体标本的血液和鳍以及幼虫的尾巴，研究了以小贝和藤条为软骨硬骨目和硬骨目代表物种的鱼类组织延长储存的潜力。对每个组织样本测试了13种延长储存的程序，这些程序在现场和实验室条件下都是为了快速和简单而选择的。在0-4℃下储存1、5和10天或在-80℃下冷冻后，流式细胞术在取样后立即应用于新鲜组织，并应用于受实验方案约束的组织，始终与物种特异性标准一起使用。荧光染料4'，6-二脒-2'-苯基吲哚二盐酸盐的激发/发射最大为358/461 nm。基于储存样本的可测量性，评估其DNA峰的直接测量变异系数以及与新鲜组织相比的荧光强度变化，提出了延长模型物种所选组织类型储存的最佳程序。©2020作者。Wiley Periodicals LLC.代表国际细胞计量学促进会出版的《细胞计量学A部分》。
关键词：Sysmex Partec 倍性分析仪；固定；保存；甾醇；藤条；血液；鳍组织；幼虫尾部组织；变异系数；荧光强度
