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Two-Photon Excitation Spectra of Various Fluorescent Proteins within a Broad Excitation Range
Two-photon excitation fluorescence laser-scanning microscopy is the preferred method for studying dynamic processes in living organ models or even in living organisms. Thanks to near-infrared and infrared excitation, it is possible to penetrate deep into the tissue, reaching areas of interest releva...
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/PMC9656010/ https://www.ncbi.nlm.nih.gov/pubmed/36362194 http://dx.doi.org/10.3390/ijms232113407 |
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author | Leben, Ruth Lindquist, Randall L. Hauser, Anja E. Niesner, Raluca Rakhymzhan, Asylkhan |
author_facet | Leben, Ruth Lindquist, Randall L. Hauser, Anja E. Niesner, Raluca Rakhymzhan, Asylkhan |
author_sort | Leben, Ruth |
collection | PubMed |
description | Two-photon excitation fluorescence laser-scanning microscopy is the preferred method for studying dynamic processes in living organ models or even in living organisms. Thanks to near-infrared and infrared excitation, it is possible to penetrate deep into the tissue, reaching areas of interest relevant to life sciences and biomedicine. In those imaging experiments, two-photon excitation spectra are needed to select the optimal laser wavelength to excite as many fluorophores as possible simultaneously in the sample under consideration. The more fluorophores that can be excited, and the more cell populations that can be studied, the better access to their arrangement and interaction can be reached in complex systems such as immunological organs. However, for many fluorophores, the two-photon excitation properties are poorly predicted from the single-photon spectra and are not yet available, in the literature or databases. Here, we present the broad excitation range (760 nm to 1300 nm) of photon-flux-normalized two-photon spectra of several fluorescent proteins in their cellular environment. This includes the following fluorescent proteins spanning from the cyan to the infrared part of the spectrum: mCerulean3, mTurquoise2, mT-Sapphire, Clover, mKusabiraOrange2, mOrange2, LSS-mOrange, mRuby2, mBeRFP, mCardinal, iRFP670, NirFP, and iRFP720. |
format | Online Article Text |
id | pubmed-9656010 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-96560102022-11-15 Two-Photon Excitation Spectra of Various Fluorescent Proteins within a Broad Excitation Range Leben, Ruth Lindquist, Randall L. Hauser, Anja E. Niesner, Raluca Rakhymzhan, Asylkhan Int J Mol Sci Article Two-photon excitation fluorescence laser-scanning microscopy is the preferred method for studying dynamic processes in living organ models or even in living organisms. Thanks to near-infrared and infrared excitation, it is possible to penetrate deep into the tissue, reaching areas of interest relevant to life sciences and biomedicine. In those imaging experiments, two-photon excitation spectra are needed to select the optimal laser wavelength to excite as many fluorophores as possible simultaneously in the sample under consideration. The more fluorophores that can be excited, and the more cell populations that can be studied, the better access to their arrangement and interaction can be reached in complex systems such as immunological organs. However, for many fluorophores, the two-photon excitation properties are poorly predicted from the single-photon spectra and are not yet available, in the literature or databases. Here, we present the broad excitation range (760 nm to 1300 nm) of photon-flux-normalized two-photon spectra of several fluorescent proteins in their cellular environment. This includes the following fluorescent proteins spanning from the cyan to the infrared part of the spectrum: mCerulean3, mTurquoise2, mT-Sapphire, Clover, mKusabiraOrange2, mOrange2, LSS-mOrange, mRuby2, mBeRFP, mCardinal, iRFP670, NirFP, and iRFP720. MDPI 2022-11-02 /pmc/articles/PMC9656010/ /pubmed/36362194 http://dx.doi.org/10.3390/ijms232113407 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 | Article Leben, Ruth Lindquist, Randall L. Hauser, Anja E. Niesner, Raluca Rakhymzhan, Asylkhan Two-Photon Excitation Spectra of Various Fluorescent Proteins within a Broad Excitation Range |
title | Two-Photon Excitation Spectra of Various Fluorescent Proteins within a Broad Excitation Range |
title_full | Two-Photon Excitation Spectra of Various Fluorescent Proteins within a Broad Excitation Range |
title_fullStr | Two-Photon Excitation Spectra of Various Fluorescent Proteins within a Broad Excitation Range |
title_full_unstemmed | Two-Photon Excitation Spectra of Various Fluorescent Proteins within a Broad Excitation Range |
title_short | Two-Photon Excitation Spectra of Various Fluorescent Proteins within a Broad Excitation Range |
title_sort | two-photon excitation spectra of various fluorescent proteins within a broad excitation range |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9656010/ https://www.ncbi.nlm.nih.gov/pubmed/36362194 http://dx.doi.org/10.3390/ijms232113407 |
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