Yang Lingling, Zhu Shaobin, Hang Wei, Wu Lina, Yan Xiaomei
Department of Chemical Biology, College of Chemistry and Chemical Engineering, The Key Laboratory for Chemical Biology of Fujian Province, Xiamen University, Xiamen 361005, China.
Anal Chem. 2009 Apr 1;81(7):2555-63. doi: 10.1021/ac802464a.
A compact, high-sensitivity, dual-channel flow cytometer (HSDCFCM) was developed for the individual analysis of nanosized particles and biomolecules. A hydrodynamic focusing technique was applied to confine the sample stream and enable small probe volume. Fluorescence bursts from single R-phycoerythrin (R-PE) molecules passing through the laser beam were well resolved from the background with signal-to-noise ratio of 17. Excellent size discrimination was demonstrated with a mixture of three sizes of polystyrene nanoparticles. Simultaneous measurement of fluorescence and light scattering signals from individual nanoparticles was demonstrated with the 100 nm fluorescent latex beads. Doxorubicin-loaded ZrO(2) nanoparticles and fluorescently stained Escherichia coli ER2738 cells were analyzed successfully with dual-channel detection. Particle counting is demonstrated with the 210 nm fluorescent latex beads, and excellent correlation (R(2) > 0.998) between the manufacturer-reported concentrations and those measured by HSDCFCM enumeration was obtained. The measured sample detection efficiency was approximately 90% on average for particle concentrations ranging from 1.62 x 10(5) to 3.93 x 10(7) particles/mL. Sample mixtures with varying proportions of fluorescently labeled and unlabeled nanoparticles were also analyzed with good ratio correspondence. By providing rapid, quantitative, and multiparameter characterization of nanoparticles, it is believed that the HSDCFCM will find many applications in the fields of bionanotechnology, bioanalytical chemistry, and biomedicine.
开发了一种紧凑、高灵敏度的双通道流式细胞仪（HSDCFCM），用于对纳米级颗粒和生物分子进行个体分析。应用了一种流体动力学聚焦技术来限制样品流并实现小的探针体积。单个藻红蛋白（R-PE）分子穿过激光束时产生的荧光猝发能够从背景中很好地分辨出来，信噪比为17。用三种尺寸的聚苯乙烯纳米颗粒混合物展示了出色的尺寸辨别能力。用100 nm荧光乳胶珠展示了对单个纳米颗粒的荧光和光散射信号的同时测量。用双通道检测成功分析了负载阿霉素的ZrO(2)纳米颗粒和荧光染色的大肠杆菌ER2738细胞。用210 nm荧光乳胶珠展示了颗粒计数，并且在制造商报告的浓度与通过HSDCFCM计数测量的浓度之间获得了出色的相关性（R(2) > 0.998）。对于浓度范围从1.62 x 10(5)到3.93 x 10(7)颗粒/mL的颗粒，平均测量的样品检测效率约为90%。还分析了具有不同比例荧光标记和未标记纳米颗粒的样品混合物，比例对应良好。通过提供对纳米颗粒的快速、定量和多参数表征，相信HSDCFCM将在生物纳米技术、生物分析化学和生物医学领域找到许多应用。
