A water cyclone to preserve insoluble aerosols in liquid flow - An interface to flow cytometry to detect airborne nucleic acid
A water cyclone to preserve insoluble aerosols in liquid flow - An interface to flow cytometry to detect airborne nucleic acid
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DOI：
10.1080/02786820802072881
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发表时间：
2008-01-01
影响因子：
5.2
通讯作者：
Hogrefe, Olga
中科院分区：
文献类型：
--
作者：
Orsini, Douglas A.;Rhoads, Kevin;Hogrefe, Olga
设计了一个微型旋风分离器，用于将细小的气溶胶缓慢地捕获到连续的液体流中。旋风分离器的几何形状设计成使得旋转空气的摩擦在锥体底部使100 μ l体积的水旋转，产生覆盖内部沉积表面的静止液体涡流。旋风分离器的收集效率的特征是不溶性颗粒尺寸的函数，无论是在独立操作中还是在水蒸气冷凝导致气溶胶增长之前。气溶胶的增长降低了最小收集颗粒的尺寸，并在撞击点处创造了同义的样品进入基底材料的条件。对于荧光聚苯乙烯胶乳珠粒直径尺寸为50-3000 nm，旋风收集效率高于88%。旋风分离器进一步与流式细胞仪连接，以检测旋风分离器样品流中的空气传播的核酸（作为病毒测试气溶胶）。流式细胞仪通常用于通过荧光进行单细胞鉴定，对流式细胞仪进行修改，以接受来自旋风分离器的连续样品流（标称60 μ l/min（-1）），用于实时检测。将杆状植物病毒（烟草花叶病毒）和蛋白质包膜昆虫病毒（杆状病毒）雾化，通过旋风分离器收集，并使用核酸染料SYBR绿色I、SYTO-9和SYTO-24（Molecular Probes，Inc.）在线染色。此外，使用环境扫描电子显微镜（ESEM）确认单个病毒颗粒的采集，并定性评价雾化和采集过程对病毒完整性的影响程度。
A miniature cyclone was designed to gently capture fine aerosols into a continuous liquid flow. The geometry of the cyclone was designed so that the friction of the turning air swirls a 100 mu l volume of water at the base of the cone, creating a standing liquid vortex which coats the inside deposition surface. The collection efficiency of the cyclone was characterized as a function of insoluble particle size, both in stand-alone operation and preceded by aerosol growth by water vapor condensation. The aerosol growth lowered the smallest collected particle size and created synonymous sample-into-substrate material conditions at the point of impact.The cyclone collection efficiencies were higher than 88% for the fluorescent polystyrene latex bead diameter sizes 50-3000 nm. The cyclone was further interfaced to a flow cytometer to detect airborne nucleic acid (as a virus test aerosol) in the cyclone sample flow. The flow cytometer, which is commonly used for single cell identification via fluorescence, was modified to accept a continuous sample flow (nominal 60 mu l min(-1)) from the cyclone for real-time detection. A rod-shaped plant virus (Tobamovirus) and a protein-enveloped insect virus (Baculovirus) were aerosolized, collected by the cyclone, and stained inline using the nucleic acid dyes SYBR Green I, SYTO-9, and SYTO-24 (Molecular Probes, Inc.). In addition, an Environmental Scanning Electron Microscope (ESEM) was used to confirm the collection of single virus particles and qualitatively evaluate the degree to which the aerosolization and collection process affected the integrity of the virus.