Self-interference 3D super-resolution microscopy for deep tissue investigations
Self-interference 3D super-resolution microscopy for deep tissue investigations
复制标题
DOI：
10.1038/s41592-018-0005-3
复制
发表时间：
2018-06-01
期刊：
影响因子：
48
通讯作者：
Cognet, Laurent
中科院分区：
文献类型：
--
作者：
Bon, Pierre;Linares-Loyez, Jeanne;Cognet, Laurent
荧光定位显微镜已达到近分子的分辨率，能够揭示超微结构，具有广泛的应用，特别是在细胞生物学。然而，在三维空间和第一细胞层以外的生物组织内部实现这种分辨率仍然具有挑战性。在这里，我们介绍SELFI，一个框架内的多细胞标本和组织的3D单分子定位。该方法依赖于显微镜的点扩散函数（PSF）内产生的自干涉，以同时编码等相位和强度荧光信号，它们一起提供发射器的3D位置。我们结合SELFI与传统的定位显微镜可视化F-肌动蛋白三维细丝网络，并揭示了空间分布的转录因子OCT 4在人类诱导多能干细胞在深度达50 μ m内未清除的组织球体。SELFI为完整组织中天然细胞过程的纳米级研究铺平了道路。
Fluorescence localization microscopy has achieved near-molecular resolution capable of revealing ultra-structures, with a broad range of applications, especially in cellular biology. However, it remains challenging to attain such resolution in three dimensions and inside biological tissues beyond the first cell layer. Here we introduce SELFI, a framework for 3D single-molecule localization within multicellular specimens and tissues. The approach relies on self-interference generated within the microscope's point spread function (PSF) to simultaneously encode equiphase and intensity fluorescence signals, which together provide the 3D position of an emitter. We combined SELFI with conventional localization microscopy to visualize F-actin 3D filament networks and reveal the spatial distribution of the transcription factor OCT4 in human induced pluripotent stem cells at depths up to 50 mu m inside uncleared tissue spheroids. SELFI paves the way to nanoscale investigations of native cellular processes in intact tissues.