Jbeily Nayla, Claus Ralf A, Dahlke Katja, Neugebauer Ute, Bauer Michael, Gonnert Falk A
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.
J Biophotonics. 2014 Jun;7(6):369-75. doi: 10.1002/jbio.201300179. Epub 2014 Jan 31.
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.
活体荧光显微镜检查（IVM）是研究白细胞募集过程中白细胞命运的理想工具。在本研究中，比较了常用于此目的的染料罗丹明6G和羧基荧光素二乙酸琥珀酰亚胺酯（CFDA-SE）在白细胞标记方面对IVM研究的适用性。通过流式细胞术评估了它们标记不同白细胞亚群的潜力以及它们的荧光强度，结果显示两种染料之间存在明显差异。这些差异对它们在白细胞-内皮细胞相互作用体内成像中的应用产生了深远影响。总之，就图像质量而言，CFDA-SE在标记用于体内显微镜检查的白细胞方面表现更优。此外，我们还能够在脓毒症动物模型的疾病条件下证明CFDA-SE的有效性。
