Two-step phase-shifting sectioning structured illumination microscopy

Hongting Wang; Zhixiong Hu; Wenli Liu; Fei Li; Xiuyu Li

doi:10.1117/12.2551040

31 January 2020 Two-step phase-shifting sectioning structured illumination microscopy

Hongting Wang, Zhixiong Hu, Wenli Liu, Fei Li, Xiuyu Li

Proceedings Volume 11427, Second Target Recognition and Artificial Intelligence Summit Forum; 1142713 (2020) https://doi.org/10.1117/12.2551040
Event: Second Target Recognition and Artificial Intelligence Summit Forum, 2019, Changchun, China

Abstract

Two-step phase-shifting sectioning structured light illumination microscopy (TSSIM) that reconstructs a three-dimensional structure using Fourier transform is proposed. Undesirable background signals corresponding to out-of-focus signals are eliminated using this method. Since there is no restriction for accurate phase shifts, this method does not suffer from large retrieval errors, unlike conventional sectioning structured illumination wide-field fluorescence microscopy (SSIWM). It can be used directly without modifying the conventional SSIWM microscope setup employing two of the three captured images, and can be applied to both shape measurements and biological observation. Less exposure time is required; thus, photobleaching and phototoxicity in biological observation are reduced. Further, the impact of the phase-shift difference on the signal-to-noise ratio of reconstruction image is analyzed. Both simulations and experiments are presented to show the validity of the proposed method.

Citation Download Citation

Hongting Wang, Zhixiong Hu, Wenli Liu, Fei Li, and Xiuyu Li "Two-step phase-shifting sectioning structured illumination microscopy", Proc. SPIE 11427, Second Target Recognition and Artificial Intelligence Summit Forum, 1142713 (31 January 2020); https://doi.org/10.1117/12.2551040

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