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Super-resolution localization microscopy with large field-of-view for mapping synaptic connectivity at multiple scales (Conference Presentation)
Author(s): Zhenli Huang; Bo Xin; Yujie Wang; Luchang Li; Zhaoning Zhang; Mingtao Shang
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Paper Abstract

Super-resolution localization microscopy (SRLM), including PALM, STORM, dSTORM and many others, achieves ultra-high spatial resolution up to 20~30 nanometers by positioning and reconstructing single molecules from thousands or even tens of thousands of raw images. As intrinsically a wide-field imaging technique, SRLM has the advantage of increasing field-of-view (FOV) without sacrificing either imaging speed or spatial resolution. Currently, limited by the number of active pixels in EMCCD cameras (typically 512 x 512), the maximum FOV of popular SRLM is approximately 50 um x 50 um at the sample plane. Such an FOV is insufficient for observing many biological phenomena which are best interpreted in large FOV, for example, volumetric mapping of synaptic connectivity at multiple scales. In this talk, we will report our recent progresses in the technology development and applications of SRLM with large FOV. We will firstly report the imaging performance of a back-illuminated sCMOS cameras with 95% QE for SRLM. Then, we discuss a high-power homogeneous illumination system which is capable of providing sufficient illumination intensity and excellent illumination homogeneity for SRLM with large FOV. Finally, we present some preliminary results of using large FOV SRLM in mapping synaptic connectivity at multiple scales.

Paper Details

Date Published: 14 March 2018
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Proc. SPIE 10481, Neural Imaging and Sensing 2018, 104810U (14 March 2018); doi: 10.1117/12.2286842
Show Author Affiliations
Zhenli Huang, Wuhan National Lab. for Optoelectronics (China)
Bo Xin, Wuhan National Lab. for Optoelectronics (China)
Yujie Wang, Wuhan National Lab. for Optoelectronics (China)
Luchang Li, Wuhan National Lab. for Optoelectronics (China)
Zhaoning Zhang, Wuhan National Lab. for Optoelectronics (China)
Mingtao Shang, Wuhan National Lab. for Optoelectronics (China)


Published in SPIE Proceedings Vol. 10481:
Neural Imaging and Sensing 2018
Qingming Luo; Jun Ding, Editor(s)

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