標(biāo)題：Sysmex Partec 倍性分析儀在魚類樣本DNA含量測量中的應(yīng)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：
倍體分析儀是一種有效且廣泛使用的魚類倍性測定工具，但并不總是能夠獲取新鮮樣本進(jìn)行分析。我們使用亞成體/成體標(biāo)本的血液和鰭以及幼蟲的尾巴，研究了以小貝和藤條為軟骨硬骨目和硬骨目代表物種的魚類組織延長儲存的潛力。對每個組織樣本測試了13種延長儲存的程序，這些程序在現(xiàn)場和實驗室條件下都是為了快速和簡單而選擇的。在0-4℃下儲存1、5和10天或在-80℃下冷凍后，流式細(xì)胞術(shù)在取樣后立即應(yīng)用于新鮮組織，并應(yīng)用于受實驗方案約束的組織，始終與物種特異性標(biāo)準(zhǔn)一起使用。熒光染料4'，6-二脒-2'-苯基吲哚二鹽酸鹽的激發(fā)/發(fā)射最大為358/461 nm?；趦Υ鏄颖镜目蓽y量性，評估其DNA峰的直接測量變異系數(shù)以及與新鮮組織相比的熒光強度變化，提出了延長模型物種所選組織類型儲存的最佳程序。?2020作者。Wiley Periodicals LLC.代表國際細(xì)胞計量學(xué)促進(jìn)會出版的《細(xì)胞計量學(xué)A部分》。
關(guān)鍵詞：Sysmex Partec 倍性分析儀；固定；保存；甾醇；藤條；血液；鰭組織；幼蟲尾部組織；變異系數(shù)；熒光強度
