Institute of Electronic Structure and Laser, Foundation for Research and Technology Hellas, GR-70013, Heraklion, Greece.
Department of Materials Science and Technology, University of Crete, GR-71003, Heraklion, Greece.
Sci Rep. 2017 Sep 19;7(1):11854. doi: 10.1038/s41598-017-12193-x.
We present a new Phase-Retrieved Tomography (PRT) method to radically improve mesoscopic imaging at regimes beyond one transport mean-free-path and achieve high resolution, uniformly throughout the volume of opaque samples. The method exploits multi-view acquisition in a hybrid Selective Plane Illumination Microscope (SPIM) and Optical Projection Tomography (OPT) setup and a three-dimensional Gerchberg-Saxton phase-retrieval algorithm applied in 3D through the autocorrelation sinogram. We have successfully applied this innovative protocol to image optically dense 3D cell cultures in the form of tumor spheroids, highly versatile models to study cancer behavior and response to chemotherapy. We have thus achieved a significant improvement of resolution in depths not yet accessible with the currently used methods in SPIM/OPT, while overcoming all registration and alignment problems inherent to these techniques.
我们提出了一种新的相衬层析成像（PRT）方法，可以在超越一个传输平均自由程的情况下，从根本上改善介观成像，并在不透明样本的整个体积中实现高分辨率、均匀的成像。该方法利用混合选择性平面照明显微镜（SPIM）和光学投影层析成像（OPT）设置中的多角度采集，以及通过自相关正弦图在 3D 中应用的三维 Gerchberg-Saxton 相位恢复算法。我们已经成功地将这种创新的方案应用于以肿瘤球体形式存在的光学密集 3D 细胞培养物成像，这些球体是研究癌症行为和对化疗反应的高度多功能模型。因此，我们在 SPIM/OPT 目前使用的方法尚无法达到的深度上实现了分辨率的显著提高，同时克服了这些技术固有的所有配准和对准问题。
