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

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 exci...

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Detalles Bibliográficos
Autores principales: Yan, Wei, Huang, Yangrui, Wang, Luwei, Li, Jin, Guo, Yong, Yang, Zhigang, Qu, Junle
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9138471/
https://www.ncbi.nlm.nih.gov/pubmed/35624608
http://dx.doi.org/10.3390/bios12050307
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author Yan, Wei
Huang, Yangrui
Wang, Luwei
Li, Jin
Guo, Yong
Yang, Zhigang
Qu, Junle
author_facet Yan, Wei
Huang, Yangrui
Wang, Luwei
Li, Jin
Guo, Yong
Yang, Zhigang
Qu, Junle
author_sort Yan, Wei
collection PubMed
description 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.
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spelling pubmed-91384712022-05-28 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 Biosensors (Basel) Communication 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. MDPI 2022-05-06 /pmc/articles/PMC9138471/ /pubmed/35624608 http://dx.doi.org/10.3390/bios12050307 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Communication
Yan, Wei
Huang, Yangrui
Wang, Luwei
Li, Jin
Guo, Yong
Yang, Zhigang
Qu, Junle
Multi-Color Two-Photon Microscopic Imaging Based on a Single-Wavelength Excitation
title Multi-Color Two-Photon Microscopic Imaging Based on a Single-Wavelength Excitation
title_full Multi-Color Two-Photon Microscopic Imaging Based on a Single-Wavelength Excitation
title_fullStr Multi-Color Two-Photon Microscopic Imaging Based on a Single-Wavelength Excitation
title_full_unstemmed Multi-Color Two-Photon Microscopic Imaging Based on a Single-Wavelength Excitation
title_short Multi-Color Two-Photon Microscopic Imaging Based on a Single-Wavelength Excitation
title_sort multi-color two-photon microscopic imaging based on a single-wavelength excitation
topic Communication
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9138471/
https://www.ncbi.nlm.nih.gov/pubmed/35624608
http://dx.doi.org/10.3390/bios12050307
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