3D refractive index tomography with quantitative oblique back-illumination microscopy
3D refractive index tomography with quantitative oblique back-illumination microscopy
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DOI：
10.1117/12.2544328
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发表时间：
2020-02
期刊：
影响因子：
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通讯作者：
P. Ledwig;Francisco E. Robles
中科院分区：
文献类型：
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作者：
P. Ledwig;Francisco E. Robles
我们最近开发了定量倾斜背照显微镜（qOBM），它能够对嵌入厚散射介质中的物体进行全场定量相位成像（QPI）。这种外延模式技术利用多重散射作为内部透射照明源，允许基于光路延迟的丰富结构细节。现在，我们通过计算整个光学系统的 3D 传递函数（包括多重散射照明）并将其从相位梯度对比度图像的垂直堆栈中解卷积，生成具有亚细胞细节的折射率的定量 3D 渲染。这种方法可以实现真正的非侵入性、无标记、断层扫描定量重建（如果包括厚组织样本在内的厚散射样本中的折射率），成本低廉且操作简单，从而使 QPI 对亚细胞结构细节的无与伦比的访问能力进入以前无法获得的研究领域。
We have recently developed quantitative oblique back-illumination microscopy (qOBM), which enables full-field quantitative phase imaging (QPI) of objects embedded in a thick scattering medium. This epi-mode technique makes use of multiple scattering as a source of transmissive illumination from within, allowing for rich structural detail based on optical path delay. We now produce quantitative 3D renderings of index of refraction with sub- cellular detail by computing a 3D transfer function of the entire optical system, including the multiply scattered illumination, and deconvolving it from a vertical stack of phase gradient contrast images. This approach allows truly non-invasive, label-free, tomographic quantitative reconstructions if index of refraction in thick scattering samples including thick tissue samples, at low cost and with simple operation, bringing QPI’s unparalleled access to sub-cellular structural detail to previously unavailable domains of investigation.