Comparative suitability of CFDA-SE and rhodamine 6G for in vivo assessment of leukocyte-endothelium interactions.
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 their application for in vivo imaging of leukocyte-endothelium interactions. In summary, CFDA-SE revealed superior in labelling leukocytes for in vivo microscopy with respect to image quality. In addition, we could show the efficiency of CFDA-SE also under disease condition in an animal model of sepsis.
Integrated Research and Treatment Center, Center for Sepsis Control and Care CSCC, Jena University Hospital, Erlanger Allee 101, 07747 Jena, Germany; International Leibniz Research School, Jena, Germany.
Nayla
N
Jbeily
Ralf A
RA
Claus
Katja
K
Dahlke
Ute
U
Neugebauer
Michael
M
Bauer
Falk A
FA
Gonnert
D000818:Animals; D002450:Cell Communication; D004727:Endothelium / Q000166:cytology / Q000473:pathology; D005240:Feasibility Studies; D005452:Fluoresceins / Q000378:metabolism*; D005456:Fluorescent Dyes / Q000378:metabolism; D007962:Leukocytes / Q000166:cytology*; D008099:Liver / Q000166:cytology / Q000473:pathology; D008297:Male; D051379:Mice; D008856:Microscopy, Fluorescence / Q000379:methods*; D012235:Rhodamines / Q000378:metabolism*; D018805:Sepsis / Q000276:immunology / Q000473:pathology; D013388:Succinimides / Q000378:metabolism*
C087165:5-(6)-carboxyfluorescein diacetate succinimidyl ester; D005452:Fluoresceins; D005456:Fluorescent Dyes; D012235:Rhodamines; D013388:Succinimides; C026188:rhodamine 6G
carboxyfluorescein diacetate succinimidyl ester (CFDA-SE); intravital microscopy; leukocyte-endothelium interaction; rhodamine 6G