Method for Multiplexed Dynamic Intravital Multiphoton Imaging.
Fluorescing proteins
Intravital imaging
Multiphoton microscopy
Near-infrared fluorescent dyes
Optical parametric oscillator
Similarity unmixing
Two-color two-photon excitation
Journal
Methods in molecular biology (Clifton, N.J.)
ISSN: 1940-6029
Titre abrégé: Methods Mol Biol
Pays: United States
ID NLM: 9214969
Informations de publication
Date de publication:
2021
2021
Historique:
entrez:
31
7
2021
pubmed:
1
8
2021
medline:
2
9
2021
Statut:
ppublish
Résumé
Intravital two-photon microscopy enables monitoring of cellular dynamics and communication of complex systems, in genuine environment-the living organism. Particularly, its application in understanding the immune system brought unique insights into pathophysiologic processes in vivo. Here we present a method to achieve multiplexed dynamic intravital two-photon imaging by using a synergistic strategy combining a spectrally broad range of fluorophore emissions, a wave-mixing concept for simultaneous excitation of all targeted fluorophores, and an effective unmixing algorithm based on the calculation of spectral similarities with previously acquired fluorophore fingerprints. Our unmixing algorithm allows us to distinguish 7 fluorophore signals corresponding to various cellular and tissue compartments by using only four detector channels.
Identifiants
pubmed: 34331284
doi: 10.1007/978-1-0716-1593-5_10
doi:
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
145-156Informations de copyright
© 2021. Springer Science+Business Media, LLC, part of Springer Nature.
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