Crawford Andrew M, Kurecka Patrick, Yim Tsz Kwan, Kozemchak Claire, Deb Aniruddha, Dostál Lubomír, Sun Cheng Jun, Brewe Dale L, Barrea Raul, Penner-Hahn James E
Department of Chemistry, University of Michigan, Ann Arbor, MI 48109-1055, USA.
X-ray Science Division, Argonne National Laboratory, 9700 South Cass Avenue, Argonne, IL 60439, USA.
J Synchrotron Radiat. 2016 Jul;23(Pt 4):901-8. doi: 10.1107/S1600577516008006. Epub 2016 Jun 17.
An X-ray fluorescence flow cytometer that can determine the total metal content of single cells has been developed. Capillary action or pressure was used to load cells into hydrophilic or hydrophobic capillaries, respectively. Once loaded, the cells were transported at a fixed vertical velocity past a focused X-ray beam. X-ray fluorescence was then used to determine the mass of metal in each cell. By making single-cell measurements, the population heterogeneity for metals in the µM to mM concentration range on fL sample volumes can be directly measured, a measurement that is difficult using most analytical methods. This approach has been used to determine the metal composition of 936 individual bovine red blood cells (bRBC), 31 individual 3T3 mouse fibroblasts (NIH3T3) and 18 Saccharomyces cerevisiae (yeast) cells with an average measurement frequency of ∼4 cells min(-1). These data show evidence for surprisingly broad metal distributions. Details of the device design, data analysis and opportunities for further sensitivity improvement are described.
一种能够测定单细胞总金属含量的X射线荧光流式细胞仪已被开发出来。分别利用毛细作用或压力将细胞加载到亲水性或疏水性毛细管中。一旦加载完成，细胞以固定的垂直速度通过聚焦的X射线束进行传输。然后利用X射线荧光来测定每个细胞中的金属质量。通过进行单细胞测量，可以直接测量fL样品体积中微摩尔到毫摩尔浓度范围内金属的群体异质性，而这一测量使用大多数分析方法都很困难。这种方法已被用于测定936个单个牛红细胞（bRBC）、31个单个3T3小鼠成纤维细胞（NIH3T3）和18个酿酒酵母细胞的金属组成，平均测量频率约为4个细胞/分钟。这些数据显示出令人惊讶的广泛金属分布的证据。文中描述了该设备的设计细节、数据分析以及进一步提高灵敏度的机会。
