少量培养细胞的定量分析

K. Sera, S. Goto, T. Hosokawa, Y. Saitoh, T. Nagamine
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引用次数: 0

摘要

本文建立了对少量培养细胞进行定量元素分析的方法。首先建立细胞浓度大于1 × 106个/mL的溶液内标法,然后建立培养细胞的无标法。经证实,该方法可在仅含2万个细胞的样品中定量分析25种以上的元素。此外,由于使用胰蛋白酶和PBS有时会带来大量的钠、磷和钾,这对基于无标准法的分析准确性有直接影响,因此,为了提高分析的准确性和灵敏度,研究了从烧瓶中去除培养细胞的方法。发现不使用胰蛋白酶和PBS用刮刀去除细胞的方法是最好的方法。此外,为了提高分析的灵敏度,研究了使用较薄的背衬材料的影响。当使用较薄的衬底材料时,可以在无标准方法的基础上对含有近2000个细胞的样品进行准确分析。
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Quantitative analysis of very small quantities of cultured cells
In this paper, methods of quantitative elemental analysis of very small quantity of cultured cells were developed. First of all, an internal-standard method for the solution containing cells whose density is more than 1 × 106 cells/mL was established and then a standard-free method for cultured cells was developed. It was confirmed that the method allows us to quantitatively analyze more than 25 elements in the samples containing only 20,000 cells. Also, the methods for removing cultured cells from a flask were examined in order to improve accuracy and sensitivity of analysis since the use of trypsin and PBS sometimes brings a large amount of sodium, phosphorus and potassium, which have direct effect upon accuracy of analysis based on the standard-free method. It was found that the method of removing cells with a scraper without using trypsin and PBS is the best manner. Also, the effects of using thinner backing materials were examined in order to improve sensitivity of analyses. It is expected that accurate analysis of samples containing nearly two thousand cells is possible on the basis of the standard-free method when using a thinner backing material.
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