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A Mach-Zender digital holographic microscope with sub-micrometer resolution for imaging and tracking of marine micro-organisms

机译:具有亚微米分辨率的Mach-Zender数字全息显微镜,用于海洋微生物的成像和跟踪

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摘要

Digital holographic microscopy is an ideal tool for investigation of microbial motility. However, most designs do not exhibit sufficient spatial resolution for imaging bacteria. In this study we present an off-axis Mach-Zehnder design of a holographic microscope with spatial resolution of better than 800 nm and the ability to resolve bacterial samples at varying densities over a 380 μm × 380 μm × 600 μm three-dimensional field of view. Larger organisms, such as protozoa, can be resolved in detail, including cilia and flagella. The instrument design and performance are presented, including images and tracks of bacterial and protozoal mixed samples and pure cultures of six selected species. Organisms as small as 1 μm (bacterial spores) and as large as 60 μm (Paramecium bursaria) may be resolved and tracked without changes in the instrument configuration. Finally, we present a dilution series investigating the maximum cell density that can be imaged, a type of analysis that has not been presented in previous holographic microscopy studies.
机译:数字全息显微镜是研究微生物运动性的理想工具。但是,大多数设计没有为细菌成像显示足够的空间分辨率。在这项研究中,我们提出了全息显微镜的离轴Mach-Zehnder设计,其空间分辨率优于800 nm,并且能够在380μm×380μm×600μm的三维场上以不同的密度分辨细菌样品。视图。较大的生物体,例如原生动物,可以详细解析,包括纤毛和鞭毛。介绍了仪器的设计和性能,包括细菌和原生动物混合样品的图像和轨迹以及六个选定物种的纯培养物。小至1μm(细菌孢子)和大至60μm(法氏囊)的生物都可以被解析和跟踪,而无需改变仪器的配置。最后,我们提出了一个稀释系列,研究可以成像的最大细胞密度,这是以前的全息显微镜研究中未曾提出过的一种分析类型。

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