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Aberration Correction to Optimize the Performance of Two-Photon Fluorescence Microscopy Using the Genetic Algorithm

Published online by Cambridge University Press:  25 January 2022

Wei Yan Open the ORCID record for Wei Yan [Opens in a new window] ,
Yangrui Huang ,
Luwei Wang ,
Yong Guo ,
Jin Li ,
Yinru Zhu ,
Zhigang Yang  and
Junle Qu
Wei Yan
Affiliation:
Center for Biomedical Photonics & College of Physics and Optoelectronic Engineering, Key Laboratory of Optoelectronic Devices and Systems of Ministry of Education and Guangdong Province, Shenzhen University, Shenzhen 518060, China
Yangrui Huang
Affiliation:
Center for Biomedical Photonics & College of Physics and Optoelectronic Engineering, Key Laboratory of Optoelectronic Devices and Systems of Ministry of Education and Guangdong Province, Shenzhen University, Shenzhen 518060, China
Luwei Wang*
Affiliation:
Center for Biomedical Photonics & College of Physics and Optoelectronic Engineering, Key Laboratory of Optoelectronic Devices and Systems of Ministry of Education and Guangdong Province, Shenzhen University, Shenzhen 518060, China
Yong Guo
Affiliation:
Center for Biomedical Photonics & College of Physics and Optoelectronic Engineering, Key Laboratory of Optoelectronic Devices and Systems of Ministry of Education and Guangdong Province, Shenzhen University, Shenzhen 518060, China
Jin Li
Affiliation:
Center for Biomedical Photonics & College of Physics and Optoelectronic Engineering, Key Laboratory of Optoelectronic Devices and Systems of Ministry of Education and Guangdong Province, Shenzhen University, Shenzhen 518060, China
Yinru Zhu
Affiliation:
Center for Biomedical Photonics & College of Physics and Optoelectronic Engineering, Key Laboratory of Optoelectronic Devices and Systems of Ministry of Education and Guangdong Province, Shenzhen University, Shenzhen 518060, China
Zhigang Yang
Affiliation:
Center for Biomedical Photonics & College of Physics and Optoelectronic Engineering, Key Laboratory of Optoelectronic Devices and Systems of Ministry of Education and Guangdong Province, Shenzhen University, Shenzhen 518060, China
Junle Qu
Affiliation:
Center for Biomedical Photonics & College of Physics and Optoelectronic Engineering, Key Laboratory of Optoelectronic Devices and Systems of Ministry of Education and Guangdong Province, Shenzhen University, Shenzhen 518060, China
*
*Corresponding author: Luwei Wang, E-mail: [email protected]
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Abstract

Due to less light scattering and a better signal-to-noise ratio in deep imaging, two-photon fluorescence microscopy (TPFM) has been widely used in biomedical photonics since its advent. However, optical aberrations degrade the performance of TPFM in terms of the signal intensity and the imaging depth and therefore restrict its application. Here, we introduce adaptive optics based on the genetic algorithm to detect the distorted wavefront of the excitation laser beam and then perform aberration correction to optimize the performance of TPFM. By using a spatial light modulator as the wavefront controller, the correction phase is obtained through a signal feedback loop and a process of natural selection. The experimental results show that the signal intensity and imaging depth of TPFM are improved after aberration correction. Finally, the method was applied to two-photon fluorescence lifetime imaging, which helps to improve the signal-to-noise ratio and the accuracy of lifetime analysis. Furthermore, the method can also be implemented in other experiments, such as three-photon microscopy, light-sheet microscopy, and super-resolution microscopy.

Keywords

adaptive opticsgenetic algorithmoptical aberrationtwo-photon microscopy
Type
Software and Instrumentation
Information
Microscopy and Microanalysis , Volume 28 , Issue 2 , April 2022 , pp. 383 - 389
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Copyright
Copyright © The Author(s), 2022. Published by Cambridge University Press on behalf of the Microscopy Society of America

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