Sun Minglu, Shao Lina, Zhang Jinrui, Zhu Youqiang, Wu Peilin, Wang Yukun, Diao Zhihui, Mu QuanQuan, Li Dayu, Wang Hongda, Xuan Li
State Key Laboratory of Applied Optics, Changchun Institute of Optics, Fine Mechanics and Physics, Chinese Academy of Sciences, Changchun 130033, China.
Center of Materials Science and Optoelectronics Engineering, University of Chinese Academy of Sciences, Beijing 100049, China.
Biomed Opt Express. 2022 Mar 14;13(4):2050-2067. doi: 10.1364/BOE.452363. eCollection 2022 Apr 1.
Fourier ptychography is a promising and flexible imaging technique that can achieve 2D quantitative reconstruction with higher resolution beyond the limitation of the system. Meanwhile, by using different imaging models, the same platform can be applied to achieve 3D refractive index reconstruction. To improve the illumination NA as much as possible while reducing the intensity attenuation problem caused by the LED board used in the traditional FP platform, we apply a hemispherical lighting structure and design a new LED arrangement according to 3D Fourier diffraction theory. Therefore, we could obtain the illumination of 0.98NA using 187 LEDs and achieve imaging half-pitch resolutions of ∼174 nm and ∼524 nm for the lateral and axial directions respectively, using a 40×/0.6NA objective lens. Furthermore, to reduce the number of captured images required and realize real-time data collection, we apply the multiplexed-coded illumination strategy and compare several coded patterns through simulation and experiment. Through comparison, we determined a radial-coded illumination pattern that could achieve more similar results as sequential scanning and increase the acquisition speed to above 1 Hz. Therefore, this paper provides the possibility of this technique in real-time 3D observation of live samples.
傅里叶叠层成像术是一种很有前景且灵活的成像技术,它能够突破系统限制,以更高分辨率实现二维定量重建。同时,通过使用不同的成像模型,同一平台可用于实现三维折射率重建。为了在减少传统傅里叶叠层成像平台中使用的LED板所导致的强度衰减问题的同时,尽可能提高照明数值孔径,我们采用了半球形照明结构,并根据三维傅里叶衍射理论设计了一种新的LED排列方式。因此,使用187个LED,我们能够获得数值孔径为0.98的照明,并使用40×/0.6NA物镜分别在横向和轴向上实现约174 nm和约524 nm的成像半间距分辨率。此外,为了减少所需采集的图像数量并实现实时数据采集,我们应用了多路复用编码照明策略,并通过模拟和实验比较了几种编码图案。通过比较,我们确定了一种径向编码照明图案,它能够实现与顺序扫描更相似的结果,并将采集速度提高到1 Hz以上。因此,本文为该技术在实时三维观察活样本方面提供了可能性。