Dojčilović Radovan, Pajović Jelena D, Božanić Dušan K, Bogdanović Una, Vodnik Vesna V, Dimitrijević-Branković Suzana, Miljković Miona G, Kaščaková Slavka, Réfrégiers Matthieu, Djoković Vladimir
Vinča Institute of Nuclear Sciences, University of Belgrade, P.O. Box 522, 11001 Belgrade, Serbia.
Faculty of Physics, University of Belgrade, P. O. Box 368, 11001 Belgrade, Serbia; DISCO Beamline, Synchrotron SOLEIL, F-91192 Gif sur Yvette, France.
Colloids Surf B Biointerfaces. 2017 Jul 1;155:341-348. doi: 10.1016/j.colsurfb.2017.04.044. Epub 2017 Apr 23.
The interaction of the tryptophan functionalized Ag nanoparticles and live Candida albicans cells was studied by synchrotron excitation deep-ultraviolet (DUV) fluorescence imaging at the DISCO beamline of Synchrotron SOLEIL. DUV imaging showed that incubation of the fungus with functionalized nanoparticles results in significant increase in the fluorescence signal. The analysis of the images revealed that the interaction of the nanoparticles with (pseudo)hyphae polymorphs of the diploid fungus was less pronounced than in the case of yeast cells or budding spores. The changes in the intensity of the fluorescence signals of the cells after incubation were followed in [327-353nm] and [370-410nm] spectral ranges that correspond to the fluorescence of tryptophan in non-polar and polar environment, respectively. As a consequence of the environmental sensitivity of the silver-tryptophan fluorescent nanoprobe, we were able to determine the possible accumulation sites of the nanoparticles. The analysis of the intensity decay kinetics showed that the photobleaching effects were more pronounced in the case of the functionalized nanoparticle treated cells. The results of time-integrated emission in the mentioned spectral ranges suggested that the nanoparticles penetrate the cells, but that the majority of the nanoparticles attach to the cells' surfaces.
通过在同步加速器SOLEIL的DISCO光束线进行同步激发深紫外(DUV)荧光成像,研究了色氨酸功能化的银纳米颗粒与活的白色念珠菌细胞之间的相互作用。DUV成像显示,用功能化纳米颗粒孵育真菌会导致荧光信号显著增加。图像分析表明,纳米颗粒与二倍体真菌的(假)菌丝多晶型物的相互作用不如与酵母细胞或出芽孢子的相互作用明显。孵育后,在分别对应于非极性和极性环境中色氨酸荧光的[327 - 353nm]和[370 - 410nm]光谱范围内跟踪细胞荧光信号强度的变化。由于银 - 色氨酸荧光纳米探针的环境敏感性,我们能够确定纳米颗粒可能的积累位点。强度衰减动力学分析表明,在功能化纳米颗粒处理的细胞中光漂白效应更明显。在上述光谱范围内的时间积分发射结果表明,纳米颗粒穿透细胞,但大多数纳米颗粒附着在细胞表面。