Automated sample preparation method for suspension arrays using renewable surface separations with multiplexed flow cytometry fluorescence detection

In this paper, we describe a new method of automated sample preparation for multiplexed biological analysis systems that use flow cytometry fluorescence detection. In this approach, color-encoded microspheres derivatized to capture particular biomolecules are temporarily trapped in a renewable surface separation column to enable perfusion with sample and reagents prior to delivery to the detector. This method provides for separation of the biomolecules of interest from other sample matrix components as well as from labeling solutions. After sample preparation, the beads can be released from the renewable surface column and delivered to a flow cytometer for direct on-bead analysis one bead at a time. Using mixtures of color-encoded beads derivatized for various analytes yields suspension arrays for multiplexed analysis. Development of this approach required a new technique for automated capture and release of the color-encoded microspheres within a fluidic system. We developed a method for forming a renewable filter and demonstrate its use for capturing microspheres that are too small to be easily captured in previous flow cells for renewable separation columns. The renewable filter is created by first trapping larger beads in the flow cell, and then smaller beads are captured either within or on top of the bed of larger beads. Both the selective microspheres and filter bed are automatically emplaced and discarded for each sample. A renewable filter created with 19.9 μm beads was used to trap 5.6 μm optically encoded beads with trapping efficiencies of 99%. The larger beads forming the renewable filter did not interfere with the detection of color-encoded 5.6 μm beads by the flow cytometer fluorescence detector. The use of this method was demonstrated with model reactions for a variety of bioanalytical assay types including a one-step capture of a biotinylated label on Lumavidin beads, a two-step sandwich immunoassay, and a one-step DNA binding assay. A preliminary demonstration of multiplexed detection of two analytes using color-encoded beads was also demonstrated. The renewable filter for creating separation columns containing optically encoded beads provides a general platform for coupling renewable surface methods for sample preparation and analyte labeling with flow cytometry detectors for suspension array multiplexed analyses.     

水溶性量子点纳米微球的制备、表征及其在生物检测中的应用

通过超声乳化(O/W)法, 在CdSe/CdS荧光量子点外包覆一层双亲性高分子外壳, 制得水溶性量子点纳米微球. 用荧光发射光谱(PL)和透射电子显微镜(TEM)等手段对产品进行了表征. 结果表明, 此种方法简单易行, 制得的量子点纳米微球(70 nm)具有良好的水溶性、稳定性以及较强的荧光发射强度. 用这种改性后的量子点标记的免疫球蛋白分子能够识别专一抗原, 因此这种纳米粒子将有望进一步应用于生物检测.     

Rolling--a new application technique for luminescent bacteria on high-performance thin-layer chromatography plates

High-performance thin-layer chromatography (HPTLC) coupled with bioluminescence detection using Vibrio fischeri bacteria can be used for screening for unknown substances. This is accomplished by dipping the HPTLC plate in an aqueous bacteria solution. Especially polar substances, however, can start to dissolve during this process, which leads to blurring and tailing of the zones on the plate. To overcome this disadvantage, we applied the bacteria solution by rolling. This method has been described for chemical derivatizations, but is very rarely used. The rolling device was made of commercially available household articles. Using octhilinone and methylparaben as test compounds, rolling was compared with dipping. Despite of performing the rolling process manually, the results were reproducible. Depending on the substance and its amount on the HPTLC plate, peaks were narrower, up to a factor of 4 higher and with a higher signal-to-noise ratio than after dipping.     

多功能金属石墨纳米囊的生物医学应用进展

多功能金属石墨纳米囊由于其良好的稳定性和独特的理化性质, 在生物医学领域受到了广泛关注. 利用石墨烯外壳独特的拉曼散射特征峰作为拉曼标签或者内标, 结合等离子体纳米核优异的表面增强拉曼散射(SERS)和双光子发光(TPL)性能, 可实现SERS生物分析以及肿瘤细胞或组织的Raman/TPL双模成像. 利用表面积大的石墨烯外壳作为药物负载平台, 结合等离子体纳米核的近红外光吸收能力, 可实现光介导的病原菌杀灭以及肿瘤细胞或实体瘤的热疗与化疗的协同治疗. 此外, 利用石墨烯外壳优异的荧光猝灭性能, 还实现了生物分子的荧光检测; 利用磁性纳米核独特的磁学性能, 可实现生物样品的分离和富集、 细菌的原位磁共振成像检测以及磁靶向胃部口服药物的递送. 本综述首先介绍了金属石墨纳米囊的制备、 分类和性质, 然后概述了它们在生物检测、 生物成像和治疗3个方面的应用进展, 并进一步总结了它们的发展现状包括生物毒性和生物医学应用的优缺点, 最后对其在生物医学领域的发展方向做出了展望. 我们期望多功能的金属石墨纳米囊能够为今后的临床生物医学应用提供可靠的纳米平台.     

Versatile metal graphitic nanocapsules for SERS bioanalysis

A comprehensive summary toward the unique properties of the novel graphitic nanomaterial of metal graphitic nanocapsules (MGNs) and their applications in SERS biodetection and bioimaging were presented here.     

Versatile metal graphitic nanocapsules for SERS bioanalysis

The novel graphitic nanomaterial of metal graphitic nanocapsules (MGNs) with superior stability, unique optical properties and biocompatibility possess great potential in biomedical and bioanalytical applications. The graphitic shell can quench the background fluorescence interference from external environments via a fluorescence resonance energy transfer (FRET) process and even avoid unnecessary reactions catalyzed by inner metal core. The graphitic shell with several characteristic Raman bands itself can act as Raman signal probe or internal standard (IS), especially the 2D-band within the cellular Raman-silent region helps to reduce the interference signals from external conditions. The present context attempts to give a comprehensive overview about the preparation and unique properties of MGNs as well as their applications in SERS biodetection and bioimaging.