Ultraviolet multi-spectral microscopy using iterative phase-recovery from chromatic aberrations
Ultraviolet multi-spectral microscopy using iterative phase-recovery from chromatic aberrations
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使用色差迭代相位恢复的紫外多光谱显微镜
DOI：
10.1117/12.2508972
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发表时间：
2019
期刊：
影响因子：
--
通讯作者：
Park, YongKeun
中科院分区：
文献类型：
--
作者：
Ojaghi, Ashkan;Robles, Francisco E.;Popescu, Gabriel;Park, YongKeun
光谱的紫外区通过提供许多重要内源性生物分子的高度特异性定量信息，为生物样品的无标记分子成像提供了独特的能力。然而，紫外光谱成像在生物医学领域的应用受到了限制。为此，我们最近推出了紫外高光谱干涉（UHI）显微镜，它适用于干涉测量，以克服与紫外光谱应用于分子成像时的重大挑战。在这里，我们提出了一种替代方法，紫外多光谱显微镜，使更快的宽视场成像的代价更少的光谱数据点。我们使用紫外敏感相机检测宽视场，并使用几个（>5个）紫外滤光片恢复光谱信息，而不是使用成像光谱仪进行线扫描来恢复高分辨率光谱信息。此外，我们不是使用干涉测量来恢复相位以校正色差，而是利用色差本身来获得离焦强度图像的堆叠（在各种波长下），然后应用强度传输（TIE）方程的迭代解来恢复相位并在所有波长下产生聚焦图像，而无需移动样品或物镜。这种配置大大简化了仪器，减少了其占地面积，使其更便宜，同时实现了快速，广域成像，具有更好的光子效率。我们通过对红细胞的一系列模拟和实验来评估这种技术的能力，这些模拟和实验显示出与UHI和血红蛋白吸收特性的良好定量一致性。潜在的生物医学应用也进行了讨论。
The ultraviolet region of the spectrum offers unique capabilities for label-free molecular imaging of biological samples by providing highly-specific, quantitative information of many important endogenous biomolecules. However, the application of UV spectral imaging to biomedicine has been limited. To this end, we have recently introduced ultraviolet hyperspectral interferometric (UHI) microscopy, which applies interferometry to overcome significant challenges associated with UV spectroscopy when applied to molecular imaging. Here we present an alternative approach for UV multi-spectral microscopy which enables faster wide-field imaging at the expense of fewer spectral data points. Instead of line-scanning to recover high-resolution spectral information with an imaging spectrometer, we detect a wide field-of-view using a UV-sensitive camera and recover the spectral information using several (>5) UV-filters. Moreover, rather than using interferometry to recover the phase to correct for chromatic aberrations, we leverage the chromatic aberrations themselves to obtain a stack of through-focus intensity images (at various wavelengths) and then apply an iterative solution of the Transport of Intensity (TIE) equation to recover the phase and produce in-focus images at all wavelengths without moving the sample or objective. This configuration greatly simplifies the instrumentation, reducing its footprint and making it less expensive, while enabling fast, wide area imaging with better photon efficiency. We assess the capabilities of this technique through a series of simulations and experiments on red blood cells, which show good quantitative agreement with UHI and tabulated hemoglobin absorption properties. Potential biomedical applications are also discussed.
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影响因子：
4.6
作者：
Ojaghi A;Fay ME;Lam WA;Robles FE
通讯作者：
Robles FE
通过低相干干涉测量法，使用折射率的实部和虚部分离散射和吸收系数。
影响因子：
3.6
作者：
Robles,FranciscoE;Wax,Adam
通讯作者：
Wax,Adam
影响因子：
48
作者：
Zeskind, Benjamin J.;Jordan, Caroline D.;Matsudaira, Paul
通讯作者：
Matsudaira, Paul
光学活检 - 用光检测癌症
DOI：
--
发表时间：
1996
期刊：
Biomedical Optical Spectroscopy and Diagnostics
影响因子：
--
作者：
A. Katz;R. Alfano
通讯作者：
R. Alfano