Division of Engineering & Applied Science, California Institute of Technology, 1200 E California Blvd, Pasadena, CA, 91125, USA.
Samsung Engineering, 26 Sangil-ro 6-gil, Gangdong-gu, Seoul, Korea.
Sci Rep. 2019 Jul 31;9(1):11114. doi: 10.1038/s41598-019-47146-z.
We report the implementation of a parallel microscopy system (96 Eyes) that is capable of simultaneous imaging of all wells on a 96-well plate. The optical system consists of 96 microscopy units, where each unit is made out of a four element objective, made through a molded injection process, and a low cost CMOS camera chip. By illuminating the sample with angle varying light and applying Fourier Ptychography, we can improve the effective brightfield imaging numerical aperture of the objectives from 0.23 to 0.3, and extend the depth of field from ±5 μm to ±15 μm. The use of Fourier Ptychography additionally allows us to computationally correct the objectives' aberrations out of the rendered images, and provides us with the ability to render phase images. The 96 Eyes acquires raw data at a rate of 0.7 frame per second (all wells) and the data are processed with 4 cores of graphical processing units (GPUs; GK210, Nvidia Tesla K80, USA). The system is also capable of fluorescence imaging (excitation = 465 nm, emission = 510 nm) at the native resolution of the objectives. We demonstrate the capability of this system by imaging S1P-eGFP-Human bone osteosarcoma epithelial (U2OS) cells.
我们报告了一种并行显微镜系统(96 眼)的实现,该系统能够同时对 96 孔板上的所有孔进行成像。光学系统由 96 个显微镜单元组成,每个单元由四个元素物镜组成,通过注塑成型工艺制成,以及低成本 CMOS 相机芯片。通过用角度变化的光照射样品并应用傅里叶叠层术,我们可以将物镜的有效明场成像数值孔径从 0.23 提高到 0.3,并将景深从±5 μm扩展到±15 μm。傅里叶叠层术的使用还允许我们从渲染图像中计算校正物镜的像差,并为我们提供渲染相图的能力。96 眼以 0.7 帧/秒(所有孔)的速率获取原始数据,并且使用 4 个图形处理单元(GPU;GK210、Nvidia Tesla K80、美国)来处理数据。该系统还能够以物镜的固有分辨率进行荧光成像(激发=465nm,发射=510nm)。我们通过对 S1P-eGFP-人骨肉瘤上皮(U2OS)细胞进行成像来证明该系统的能力。