Measurement of quantity of iron in magnetically labeled cells: comparison among different UV/VIS spectrometric methods
Measurement of quantity of iron in magnetically labeled cells: comparison among different UV/VIS spectrometric methods
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DOI：
10.2144/000112599
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发表时间：
2007-11-01
期刊：
影响因子：
2.7
通讯作者：
Arbab, Ali S.
中科院分区：
文献类型：
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作者：
Rad, Ali M.;Janic, Branislava;Arbab, Ali S.
用超顺磁性氧化铁（SPIO）标记细胞正在成为细胞磁共振成像（MRI）中的常规程序。定量标记细胞中的细胞内铁是通过定量MRI研究确定累积细胞数量的先决条件。为了建立最灵敏和可重复的方法，用于测量磁标记细胞中的铁浓度，我们使用紫外-可见（UV/维斯）分光光度计研究和比较了四种不同的方法。获得了5和10 M盐酸，100 mM柠檬酸加抗坏血酸和红菲咯啉磺酸盐（BPS）的混合物的背景光谱，抗5 M盐酸加5%亚铁氰化物的混合物。还产生了含有10或5 μ g/mL氧化铁的相同溶液的光谱，以确定溶解铁的峰值吸收波长。此外，使用不同的已知铁浓度来获得每种方法的校准线。基于校准因子，在具有已知量的铁的样品和标记的细胞中测量铁。基于使用10 M盐酸的方法低估了所有实验中的铁浓度;为了使该方法得到准确的测量，样品中的铁浓度需要至少为3 μ g/mL。
Cell labeling with superparamagnetic iron oxides (SPIO) is becoming a routine procedure in cellular magnetic resonance imaging (MRI). Quantifying the intracellular iron in labeled cells is a prerequisite for determining the number of accumulated cells by quantitative MRI studies. To establish the most sensitive and reproducible method,for measuring iron concentration in magnetically labeled cells, we investigated and compared four different methods using an ultraviolet-visible (UV/VIS) spectrophotometer Background spectra were obtained for 5 and 10 M hydrochloric acids, a mixture of 100 mM citric acid plus ascorbic acid and bathophenanthroline sulphonate (BPS), anti a mixture of 5 M hydrochloric acid plus 5% ferrocyanide. Spectra of the same solutions containing either 10 or 5 mu g/mL iron oxides were also created to determine the peak absorbance wavelengths for the dissolved iron. In addition, different known iron concentrations were used to obtain calibration lines for each method. Based on the calibration factors, iron was measured in samples with a known amount of iron and in labeled cells. Methods based on the use of 10 M hydrochloric acid underestimated iron concentration in all experiments; for this method to give an accurate measurement, iron concentration in sample needs to be at least 3 mu g/mL.