title: Fast super-resolution imaging with ultra-high labeling density achieved by joint tagging super-resolution optical fluctuation imaging.
abstract: Previous stochastic localization-based super-resolution techniques are largely limited by the labeling density and the fidelity to the morphology of specimen. We report on an optical super-resolution imaging scheme implementing joint tagging using multiple fluorescent blinking dyes associated with super-resolution optical fluctuation imaging (JT-SOFI), achieving ultra-high labeling density super-resolution imaging. To demonstrate the feasibility of JT-SOFI, quantum dots with different emission spectra were jointly labeled to the tubulin in COS7 cells, creating ultra-high density labeling. After analyzing and combining the fluorescence intermittency images emanating from spectrally resolved quantum dots, the microtubule networks are capable of being investigated with high fidelity and remarkably enhanced contrast at sub-diffraction resolution. The spectral separation also significantly decreased the frame number required for SOFI, enabling fast super-resolution microscopy through simultaneous data acquisition. As the joint-tagging scheme can decrease the labeling density in each spectral channel, thereby bring it closer to single-molecule state, we can faithfully reconstruct the continuous microtubule structure with high resolution through collection of only 100 frames per channel. The improved continuity of the microtubule structure is quantitatively validated with image skeletonization, thus demonstrating the advantage of JT-SOFI over other localization-based super-resolution methods.
authors: Department of Biomedical Engineering, College of Engineering, Peking University, Beijing, China.
authors: Zhiping
authors: Z
authors: Zeng
authors: Department of Biomedical Engineering, College of Engineering, Peking University, Beijing, China.
authors: Xuanze
authors: X
authors: Chen
authors: Department of Biomedical Engineering, College of Engineering, Peking University, Beijing, China.
authors: Hening
authors: H
authors: Wang
authors: College of Life Sciences, Peking University, Beijing, China.
authors: Ning
authors: N
authors: Huang
authors: College of Life Sciences, Peking University, Beijing, China.
authors: Chunyan
authors: C
authors: Shan
authors: Department of Biomedical Engineering, College of Engineering, Peking University, Beijing, China.
authors: Hao
authors: H
authors: Zhang
authors: College of Life Sciences, Peking University, Beijing, China.
authors: Junlin
authors: J
authors: Teng
authors: Department of Biomedical Engineering, College of Engineering, Peking University, Beijing, China.
authors: Peng
authors: P
authors: Xi