Kočišová Eva, Praus Petr, Bok Jiří, Bonneau Stéphanie, Sureau Franck
Faculty of Mathematics and Physics, Institute of Physics, Charles University in Prague, 12116, Prague 2, Czech Republic.
Laboratoire Jean Perrin, Université Pierre et Marie Curie, case courrier 114, 4 Place Jussieu, 75005, Paris, France.
J Fluoresc. 2015 Sep;25(5):1245-50. doi: 10.1007/s10895-015-1612-3. Epub 2015 Jul 16.
Time-resolved microspectrofluorimetry and fluorescence microscopy imaging-two complementary fluorescence techniques-provide important information about the intracellular distribution, level of uptake and binding/interactions inside living cell of the labeled molecule of interest. They were employed to monitor the "fate" of AS1411 aptamer labeled by ATTO 425 in human living cells. Confocal microspectrofluorimeter adapted for time-resolved intracellular fluorescence measurements by using a phase-modulation principle with homodyne data acquisition was employed to obtain emission spectra and to determine fluorescence lifetimes in U-87 MG tumor brain cells and Hs68 non-tumor foreskin cells. Acquired spectra from both the intracellular space and the reference solutions were treated to observe the aptamer localization and its interaction with biological structures inside the living cell. The emission spectra and the maximum emission wavelengths coming from the cells are practically identical, however significant lifetime lengthening was observed for tumor cell line in comparison to non-tumor one.
时间分辨显微光谱荧光测定法和荧光显微镜成像——两种互补的荧光技术——提供了有关目标标记分子在活细胞内的细胞内分布、摄取水平以及结合/相互作用的重要信息。它们被用于监测在人类活细胞中由ATTO 425标记的AS1411适配体的“命运”。使用一种采用零差数据采集的相位调制原理、适用于时间分辨细胞内荧光测量的共聚焦显微光谱荧光计，来获取发射光谱并测定U - 87 MG肿瘤脑细胞和Hs68非肿瘤包皮细胞中的荧光寿命。对从细胞内空间和参比溶液获得的光谱进行处理，以观察适配体在活细胞内的定位及其与生物结构的相互作用。来自细胞的发射光谱和最大发射波长实际上是相同的，然而与非肿瘤细胞系相比，肿瘤细胞系中观察到了显著的寿命延长。
