Computational illumination for high-speed in vitro Fourier ptychographic microscopy
Computational illumination for high-speed in vitro Fourier ptychographic microscopy
复制标题
DOI：
10.1364/optica.2.000904
复制
发表时间：
2015-10-20
期刊：
影响因子：
10.4
通讯作者：
Waller, Laura
中科院分区：
文献类型：
--
作者：
Tian, Lei;Liu, Ziji;Waller, Laura
我们展示了一种新的计算照明技术，该技术实现了大的空间-带宽-时间积，用于体外未染色的活体样品的定量相位成像。显微镜镜头可以有大视场(FOV)，也可以有高分辨率，但不能两者兼有。傅里叶变换显微技术是一种新的计算成像技术，它通过融合不同照度下拍摄的多幅图像的信息来绕过这一限制。其结果是具有宽视场和高分辨率(即大空间带宽乘积)的十亿像素规模的图像。显微显微技术具有革命性的巨大潜力，并已在数字病理学中得到应用。然而，它存在获取时间长(大约几分钟)的问题，从而限制了吞吐量。更快的捕捉时间不仅可以提高成像速度，还可以研究运动伪影会降低结果的实时样本。与固定的(例如，病理的)玻片不同，活的样本在不同的空间和时间尺度上不断演变。在这里，我们提出了一种新的信源编码方案，并结合实时硬件控制，在亚秒捕获时间的4x视场中实现了0.8NA的分辨率。我们提出了一种改进的算法和一种新的初始化方案，允许在长时间推移的实验中进行稳健的相位重建。我们提出了在体外细胞培养中生长和融合的第一个FPM结果，捕捉了流行的细胞系在经历分裂和迁移过程中的亚细胞动力学现象的视频。我们的方法将FPM应用于实时样本，用于观察空间和时间中的罕见事件。(C)2015年美国光学学会
We demonstrate a new computational illumination technique that achieves a large space-bandwidth-time product, for quantitative phase imaging of unstained live samples in vitro. Microscope lenses can have either a large field of view (FOV) or high resolution, and not both. Fourier ptychographic microscopy (FPM) is a new computational imaging technique that circumvents this limit by fusing information from multiple images taken with different illumination angles. The result is a gigapixel-scale image having both a wide FOV and high resolution, i.e., a large space-bandwidth product. FPM has enormous potential for revolutionizing microscopy and has already found application in digital pathology. However, it suffers from long acquisition times (of the order of minutes), limiting throughput. Faster capture times would not only improve the imaging speed, but also allow studies of live samples, where motion artifacts degrade results. In contrast to fixed (e.g., pathology) slides, live samples are continuously evolving at various spatial and temporal scales. Here, we present a new source coding scheme, along with real-time hardware control, to achieve 0.8 NA resolution across a 4x FOV with subsecond capture times. We propose an improved algorithm and a new initialization scheme, which allow robust phase reconstruction over long time-lapse experiments. We present the first FPM results for both growing and confluent in vitro cell cultures, capturing videos of subcellular dynamical phenomena in popular cell lines undergoing division and migration. Our method opens up FPM to applications with live samples, for observing rare events in both space and time. (C) 2015 Optical Society of America