Multi‐color two‐photon scanning structured illumination microscopy imaging of live cells

Author:

Wang Meiting1ORCID,Chen Jiajie2ORCID,Wu Wenshuai2,Wang Lei2,Zheng Xiaomin2,Xu Gaixia1,Qu Junle2ORCID,Gao Bruce Zhi3,Shao Yonghong2

Affiliation:

1. Guangdong Key Laboratory for Biomedical Measurements and Ultrasound Imaging, National‐Regional Key Technology Engineering Laboratory for Medical Ultrasound, School of Biomedical Engineering Shenzhen University Shenzhen China

2. Key Laboratory of Optoelectronic Devices and Systems of the Ministry of Education and Guangdong Province, College of Physics and Optoelectronic Engineering Shenzhen University Shenzhen China

3. Department of Bioengineering and COMSET Clemson University Clemson South Carolina USA

Abstract

AbstractMulti‐color two‐photon microscopy imaging of live cells is essential in biology. However, the limited diffraction resolution of conventional two‐photon microscopy restricts its application to subcellular organelle imaging. Recently, we developed a laser scanning two‐photon non‐linear structured illumination microscope (2P‐NLSIM), whose resolution improved three‐fold. However, its ability to image polychromatic live cells under low excitation power has not been verified. Here, to improve the reconstruction super‐resolution image quality under low excitation power, we increased the image modulation depth by multiplying the raw images with the reference fringe patterns in the reconstruction process. Simultaneously, we optimized the 2P‐NLSIM system to image live cells, including the excitation power, imaging speed, and field of view. The proposed system could provide a new imaging tool for live cells.

Publisher

Wiley

Subject

General Physics and Astronomy,General Engineering,General Biochemistry, Genetics and Molecular Biology,General Materials Science,General Chemistry

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