基于objective-type 全内反射暗场显微镜的无标记单颗粒流感病毒成像。
Label-free single-particle imaging of the influenza virus by objective-type total internal reflection dark-field microscopy.
机构信息
Department of Applied Chemistry, School of Engineering, The University of Tokyo, Tokyo, Japan.
出版信息
PLoS One. 2012;7(11):e49208. doi: 10.1371/journal.pone.0049208. Epub 2012 Nov 15.
Abstract
Here we report label-free optical imaging of single particles of the influenza virus attached on a glass surface with a simple objective-type total internal reflection dark-field microscopy (TIRDFM). The capability of TIRDFM for the imaging of single viral particles was confirmed from fine correlation of the TIRDFM images with fluorescent immunostaining image and scanning electron microscopy image. The density of scattering spots in the TIRDFM images showed a good linearity against the virus concentration, giving the limit of detection as 1.2×10(4) plaque-forming units per milliliter. Our label-free optical imaging method does require neither elaborated sample preparation nor complex optical systems, offering a good platform for rapid and sensitive counting of viral particles.
摘要
我们在此报告了一种简单的基于全内反射暗场显微镜（TIRDFM）的方法，用于对附着在玻璃表面的单个流感病毒粒子进行无标记的光学成像。通过 TIRDFM 图像与荧光免疫染色图像和扫描电子显微镜图像的精细相关性，证实了 TIRDFM 对单个病毒粒子成像的能力。TIRDFM 图像中的散射斑点密度与病毒浓度呈良好的线性关系，检测限为 1.2×10(4)噬菌斑形成单位/毫升。我们的无标记光学成像方法既不需要复杂的样品制备，也不需要复杂的光学系统，为快速、灵敏地计数病毒粒子提供了一个良好的平台。
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