Publikationsdatenbank

Method for multiplexed dynamic intravital multiphoton imaging (2021)

Art

Buchbeitrag

Autoren

Rakhymzhan, Asylkhan

Acs, Andreas

Leben, Ruth

Winkler, Thomas H.

Hauser, Anja E.

Niesner, Raluca A. (WE 2)

Quelle

Multiplexed imaging : methods and protocols — Eli Zamir (Hrsg.)

1. Auflage

New York, NY: Humana Press, 2021. Methods in molecular biology ; 2350 — S. 145–156

ISBN: 978-1-0716-1592-8

Sprache

Englisch

Verweise

URL (Volltext): https://link.springer.com/protocol/10.1007/978-1-0716-1593-5_10

DOI: 10.1007/978-1-0716-1593-5_10

Kontakt

Institut für Veterinär-Physiologie

Oertzenweg 19 b
14163 Berlin
+49 30 838 62600
physiologie@vetmed.fu-berlin.de

Abstract / Zusammenfassung

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.