通过扫描图案照明实现超分辨率双光子显微镜。
Super-resolution two-photon microscopy via scanning patterned illumination.
作者信息
Urban Ben E, Yi Ji, Chen Siyu, Dong Biqin, Zhu Yongling, DeVries Steven H, Backman Vadim, Zhang Hao F
机构信息
Department of Biomedical Engineering, Northwestern University, Evanston, Illinois, USA.
Department of Ophthalmology, Northwestern University, Chicago, Illinois, USA.
出版信息
Phys Rev E Stat Nonlin Soft Matter Phys. 2015 Apr;91(4):042703. doi: 10.1103/PhysRevE.91.042703. Epub 2015 Apr 7.
Abstract
We developed two-photon scanning patterned illumination microscopy (2P-SPIM) for super-resolution two-photon imaging. Our approach used a traditional two-photon microscopy setup with temporally modulated excitation to create patterned illumination fields. Combing nine different illuminations and structured illumination reconstruction, super-resolution imaging was achieved in two-photon microscopy. Using 2P-SPIM we achieved a lateral resolution of 141 nm, which represents an improvement by a factor of 1.9 over the corresponding diffraction limit. We further demonstrated super-resolution cellular imaging by 2P-SPIM to image actin cytoskeleton in mammalian cells and three-dimensional imaging in highly scattering retinal tissue.
摘要
我们开发了用于超分辨率双光子成像的双光子扫描图案照明显微镜（2P-SPIM）。我们的方法是在传统双光子显微镜设置中采用时间调制激发来创建图案化照明场。通过结合九种不同的照明和结构照明重建，在双光子显微镜中实现了超分辨率成像。使用2P-SPIM，我们实现了141 nm的横向分辨率，相较于相应的衍射极限提高了1.9倍。我们进一步通过2P-SPIM展示了超分辨率细胞成像，以对哺乳动物细胞中的肌动蛋白细胞骨架进行成像，并在高度散射的视网膜组织中进行三维成像。
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