Three dimensional super-resolved live cell imaging through polarized multi-angle TIRF
Posted on 2018-03-15 - 20:21
Measuring three dimension nanoscale cellular structures is challenging, especially when the structure is dynamic. Owing to the informative total internal reflection fluorescence (TIRF) imaging under varied illumination angles, multi-angle (MA) TIRF has been examined to offer a nanoscale axial and a sub-second temporal resolution. However, conventional MA-TIRF still performs badly in lateral resolution and fails to characterize the depth-image in densely-distributed regions. Here, we for the first time, emphasize the lateral super-resolution in the MA-TIRF and exampled by simply introducing polarization modulation into the illumination procedure. Equipped with a sparsity and accelerated proximal algorithm, we examine a more precise 3D sample structure compared with previous methods, enabling live cell imaging with temporal resolution of 2 seconds, recovering high-resolution mitochondria fission and fusion process. Since the introduced vortex half wave retarder is an add-on component to the existing MA-TRIF system and the algorithm is the first open sourced, we anticipate that the method and algorithm introduced here would be adopted rapidly by the biological community.
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Kuang, Cuifang; Zheng, Cheng; Zhao, Guangyuan; Liu, Wenjie; Chen, Youhua; Zhang, Zhimin; et al. (2018). Three dimensional super-resolved live cell imaging through polarized multi-angle TIRF. Optica Publishing Group. Collection. https://doi.org/10.6084/m9.figshare.c.3965565.v1
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AUTHORS (9)
CK
Cuifang Kuang
CZ
Cheng Zheng
GZ
Guangyuan Zhao
WL
Wenjie Liu
YC
Youhua Chen
ZZ
Zhimin Zhang
LJ
Luhong Jin
XL
Xu Liu
YX
Yingke Xu