High-contrast multifocus microscopy with a single camera and z-splitter prism.

High-contrast multifocus microscopy with a single camera and z-splitter prism.
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高对比度多焦点显微镜,具有单相机和z分光棱镜。

DOI:
10.1364/optica.404678
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发表时间:
2020-11-20
期刊:
影响因子:
10.4
通讯作者:
Mertz J
Mertz J
中科院分区:
物理与天体物理1区
文献类型:
--
作者:
Xiao S;Gritton H;Tseng HA;Zemel D;Han X;Mertz J

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光学显微镜一直是研究复杂生物系统的不可或缺的工具,但在进行3D体积成像时经常受到速度和复杂性问题的阻碍。在这里,我们提出了一个多焦点成像策略的基础上使用一个简单的Z-分光棱镜,可以从现成的组件组装。我们的技术使一个宽视场的图像堆栈被分配到一个单一的相机,并同时记录。我们利用我们的图像采集的体积性质,通过进一步引入一种新的扩展体积3D去卷积策略来抑制远焦荧光背景,以显着提高我们记录的图像的对比度,赋予我们的系统准光学切片的能力。通过交换不同的z分束器配置,我们可以根据感兴趣的应用优先考虑高速或大3D视场成像。此外,我们的系统可以很容易地应用到各种成像方式,除了荧光,如相衬和暗场成像。由于其简单性,多功能性和性能,我们相信我们的系统将是一个有用的工具,一般的生物或生物医学成像应用。
Optical microscopy has been an indispensable tool for studying complex biological systems, but is often hampered by problems of speed and complexity when performing 3D volumetric imaging. Here, we present a multifocus imaging strategy based on the use of a simple z-splitter prism that can be assembled from off-the-shelf components. Our technique enables a widefield image stack to be distributed onto a single camera and recorded simultaneously. We exploit the volumetric nature of our image acquisition by further introducing a novel extended-volume 3D deconvolution strategy to suppress far-out-of-focus fluorescence background to significantly improve the contrast of our recorded images, conferring to our system a capacity for quasi-optical sectioning. By swapping in different z-splitter configurations, we can prioritize high speed or large 3D field-of-view imaging depending on the application of interest. Moreover, our system can be readily applied to a variety of imaging modalities in addition to fluorescence, such as phase-contrast and darkfield imaging. Because of its simplicity, versatility, and performance, we believe our system will be a useful tool for general biological or biomedical imaging applications.
提高神经元群体记录的数据质量。
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影响因子: 25
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