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首页> 外文期刊>Cell biology international. >The application of image cytometry to viability assessment in dual fluorescence-stained fish spermatozoa.
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The application of image cytometry to viability assessment in dual fluorescence-stained fish spermatozoa.

机译：图像细胞仪在双荧光染色鱼精子活力评估中的应用。

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The viability of spermatozoa has been assessed using SYBR 14 staining for DNA of living cells and propidium iodide staining for DNA of degenerate cells. This dual staining was performed on four fish species (Siberian sturgeon, Acipenser baerii; common carp, Cyprinus carpio; tench, Tinca tinca and wels, Silurus glanis) and the proportions of live and dead spermatozoa were assessed by epifluorescence microscopy and image cytometry. Ten phase contrast and epifluorescent images were recorded per sample, corresponding images were overlaid, and the blended images were evaluated for live and dead spermatozoa, represented by green and red fluorescence signals. Live/dead proportions were assessed, after dual thresholding, by imaging software that counted absolute numbers of objects and computed their frequencies. All sperm heads were found to be labelled, emitting either green or red light. Mean numbers of spermatozoa per image were in the ranges 32-113, 61-105, 48-104 and 29-91 for Siberian sturgeon, common carp, tench and wels, respectively. The corresponding proportions of live spermatozoa were in the ranges 83.56-94.59%, 93.92-97.02%, 76.14-97.76% and 79.45-83.76%. Standard deviations did not exceed 5% of the means. The image cytometric system using dual staining with SYBR 14 and propidium iodide was clearly suitable for assessing the viability of freshwater fish spermatozoa.
机译：已使用SYBR 14染色检测了活细胞的DNA，碘化丙锭染色了了变性细胞的DNA，从而评估了精子的生存能力。这种双重染色是对四种鱼类（西伯利亚st鱼，A鱼;普通鲤鱼，鲤鱼;丁鱼，丁卡和韦尔斯，Silurus glanis）进行的，并通过落射荧光显微镜和图像细胞术评估了活和死精子的比例。每个样品记录十个相衬对比度和落射荧光图像，覆盖相应的图像，并评估混合图像的活和死精子，以绿色和红色荧光信号表示。在双重阈值之后，通过成像软件对生/死比例进行评估，该软件对物体的绝对数量进行计数并计算其频率。发现所有精子头均已标记，发出绿色或红色光。每个图像的平均精子数量在西伯利亚st鱼，鲤鱼，鲤鱼和威尔士的32-113、61-105、48-104和29-91范围内。活精子的相应比例在83.56-94.59％，93.92-97.02％，76.14-97.76％和79.45-83.76％之间。标准偏差不超过平均值的5％。使用SYBR 14和碘化丙锭双重染色的图像细胞仪系统显然适用于评估淡水鱼精子的生存力。

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