Multi-Color Two-Photon Microscopic Imaging Based on a Single-Wavelength Excitation

Yan, Wei; Huang, Yangrui; Wang, Luwei; Li, Jin; Guo, Yong; Yang, Zhigang; Qu, Junle

doi:10.3390/bios12050307

Open AccessCommunication

Multi-Color Two-Photon Microscopic Imaging Based on a Single-Wavelength Excitation

by

Wei Yan

,

Yangrui Huang

,

Luwei Wang

^*,

Jin Li

,

Yong Guo

,

Zhigang Yang

and

Junle Qu

Key Laboratory of Optoelectronic Devices and Systems of Ministry of Education and Guangdong Province, College of Physics and Optoelectronic Engineering, Shenzhen University, Shenzhen 518060, China

^*

Author to whom correspondence should be addressed.

Biosensors 2022, 12(5), 307; https://doi.org/10.3390/bios12050307

Submission received: 7 April 2022 / Revised: 2 May 2022 / Accepted: 4 May 2022 / Published: 6 May 2022

(This article belongs to the Special Issue Optical Imaging and Biophotonic Sensors (OIBS))

Download

Browse Figures

Versions Notes

Abstract

Two-photon probes with broad absorption spectra are beneficial for multi-color two-photon microscopy imaging, which is one of the most powerful tools to study the dynamic processes of living cells. To achieve multi-color two-photon imaging, multiple lasers and detectors are usually required for excitation and signal collection, respectively. However, one makes the imaging system more complicated and costly. Here, we demonstrate a multi-color two-photon imaging method with a single-wavelength excitation by using a signal separation strategy. The method can effectively solve the problem of spectral crosstalk by selecting a suitable filter combination and applying image subtraction. The experimental results show that the two-color and three-color two-photon imaging are achieved with a single femtosecond laser. Furthermore, this method can also be combined with multi-photon imaging technology to reveal more information and interaction in thick biological tissues.

Keywords: fluorescence microscopy; two-photon microscopy; multi-color imaging; signal separation

Share and Cite

MDPI and ACS Style

Yan, W.; Huang, Y.; Wang, L.; Li, J.; Guo, Y.; Yang, Z.; Qu, J. Multi-Color Two-Photon Microscopic Imaging Based on a Single-Wavelength Excitation. Biosensors 2022, 12, 307. https://doi.org/10.3390/bios12050307

AMA Style

Yan W, Huang Y, Wang L, Li J, Guo Y, Yang Z, Qu J. Multi-Color Two-Photon Microscopic Imaging Based on a Single-Wavelength Excitation. Biosensors. 2022; 12(5):307. https://doi.org/10.3390/bios12050307

Chicago/Turabian Style

Yan, Wei, Yangrui Huang, Luwei Wang, Jin Li, Yong Guo, Zhigang Yang, and Junle Qu. 2022. "Multi-Color Two-Photon Microscopic Imaging Based on a Single-Wavelength Excitation" Biosensors 12, no. 5: 307. https://doi.org/10.3390/bios12050307

APA Style

Yan, W., Huang, Y., Wang, L., Li, J., Guo, Y., Yang, Z., & Qu, J. (2022). Multi-Color Two-Photon Microscopic Imaging Based on a Single-Wavelength Excitation. Biosensors, 12(5), 307. https://doi.org/10.3390/bios12050307

Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.

Article Menu

Multi-Color Two-Photon Microscopic Imaging Based on a Single-Wavelength Excitation

Abstract

Share and Cite

Article Metrics

Article Access Statistics

Further Information

Guidelines

MDPI Initiatives

Follow MDPI