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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...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2022
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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. |
format | Online Article Text |
id | pubmed-9138471 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
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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