Computational structured illumination for high-content fluorescence and phase microscopy.
Computational structured illumination for high-content fluorescence and phase microscopy.
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DOI：
10.1364/boe.10.001978
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发表时间：
2018-12
影响因子：
3.4
通讯作者：
Li-Hao Yeh;Shwetadwip Chowdhury;L. Waller
中科院分区：
文献类型：
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作者：
Li-Hao Yeh;Shwetadwip Chowdhury;L. Waller
高内涵生物显微镜的目标是在大视场（FOV）内进行高分辨率成像。最近的工作表明，计算成像可以为高内容显微镜提供有效的解决方案。在这里，我们使用散斑结构照明显微镜（SIM）作为一个强大的和具有成本效益的解决方案，用于高含量的荧光显微镜，同时高含量的定量相位（QP）。这种多模式兼容性对于需要交叉相关生物学分析的研究至关重要。我们的方法使用横向平移的透明胶带来生成跨大FOV的高分辨率散斑照明图案。然后，定制优化算法联合重建样品的超分辨率荧光（非相干）和QP（相干）分布，同时对系统缺陷进行数字校正，例如未知的散斑照明图案、系统像差和图案平移。除了以前的线性SIM工作，我们实现了4倍的物镜的衍射限制的原生分辨率的分辨率增益，导致700 nm的荧光和1.2 μm的QP分辨率，在2 × 2.7 mm 2的FOV，给出了60兆像素的空间带宽积（SBP）。
High-content biological microscopy targets high-resolution imaging across large fields-of-view (FOVs). Recent works have demonstrated that computational imaging can provide efficient solutions for high-content microscopy. Here, we use speckle structured illumination microscopy (SIM) as a robust and cost-effective solution for high-content fluorescence microscopy with simultaneous high-content quantitative phase (QP). This multi-modal compatibility is essential for studies requiring cross-correlative biological analysis. Our method uses laterally-translated Scotch tape to generate high-resolution speckle illumination patterns across a large FOV. Custom optimization algorithms then jointly reconstruct the sample's super-resolution fluorescent (incoherent) and QP (coherent) distributions, while digitally correcting for system imperfections such as unknown speckle illumination patterns, system aberrations and pattern translations. Beyond previous linear SIM works, we achieve resolution gains of 4× the objective's diffraction-limited native resolution, resulting in 700 nm fluorescence and 1.2 μm QP resolution, across a FOV of 2 × 2.7 mm 2 , giving a space-bandwidth product (SBP) of 60 megapixels.