Monitoring the first line of defence: rhodamine 6 G and CFDA-SE and their suitability for in-vivo assessment of leukocyte-endothelium interactions

in: Journal of Biophotonics (2014)
Claus, Ralf A.; Jbeily, Nayla; Neugebauer, Ute; Bauer, Michael; Gonnert, Falk A.; Dahlke, Katja
Intravital fluorescence microscopy (IVM) is a predestined tool for investigating the fate of leukocytes during the process of leukocyte recruitment. In the present study, the commonly used dye for this purpose, rhodamine 6G, and carboxyfluorescein diacetate succinimidyl ester (CFDA-SE) were compared for leukocytes labelling with respect to suitability for IVM studies. Their potential in labelling different leukocytes subpopulations as well as their fluorescence intensities were assessed by flow cytometry revealing distinct differences between both dyes. These differences had a profound impact on its application for in-vivo imaging of leukocyte-endothelium interactions. In summary, CFDA-SE revealed superior to rhodamine 6G in the analysis of leukocyte-endothelium interactions by IVM due to a higher specificity in labelling leukocytes as well as an improved quality in imaging. In addition, we could show the efficiency of CFDA-SE also under disease condition in an animal model of peritonitis

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