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Flow cytometric assessment of cell viability: a multifaceted analysis

机译:流式细胞术评估细胞活力:多方面的分析

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Flow cytometry offers the possibility to simultaneously analyze, on a cell by cell basis, different parameters related to cell viability i.e. cell size, morphology and incorporation of dyes. Different types of analysis: light absorption of unstained/stained cells, forward angle light scattering (FALS), right angle light scattering (RALS) or both, cell fluorescence based on dye retention or dye exclusion (due to erythrosin B, ethidium bromide, fluorescein diacetate, rhodamine 123) were tested and compared, with the classical Trypan blue exclusion test, for their effectiveness in the determination of cell viability. Two types of cells in monolayer cultures (L929, SIRC) and a freshly isolated suspension of mouse splenocytes were used. For each dye, the optimal dose, incubation time and conditions for analysis were determined. Viability indications by different techniques for the three type of cell line and their reliability as compared with Trypan blue were analyzed.
机译:流式细胞术提供了在逐个细胞的基础上同时分析与细胞活力有关的不同参数的可能性,即细胞大小,形态和染料掺入。不同类型的分析:未染色/染色细胞的光吸收,前向角光散射(FALS),直角光散射(RALS)或两者兼有,基于染料保留或染料排斥的细胞荧光(由于赤藓红B,溴化乙锭,荧光素测试了双乙酸酯,若丹明123)的含量,并与经典的台盼蓝排除法进行了比较,以确定它们在确定细胞活力方面的有效性。使用单层培养物中的两种类型的细胞(L929,SIRC)和新鲜分离的小鼠脾细胞悬液。对于每种染料,确定了最佳剂量,孵育时间和分析条件。分析了三种细胞系通过不同技术的存活力指示以及与台盼蓝相比的可靠性。

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