Fixler D, Tirosh R, Eisenthal A, Lalchuk S, Marder O, Irlin Y, Deutsch M
J Biomed Opt. 1998 Jul;3(3):312-25. doi: 10.1117/1.429858.
The aim of the present study was to detect prelytic intracellular changes induced in target and effector cells following their conjugation at room temperature. Changes in the cytoplasmic matrix were measured by means of intracellular fluorescein fluorescence polarization (IFFP) using the Cellscan apparatus. Both natural killer and lymphocyte activated killer cells were used as effector cells, while K562 and Daudi cell lines were used as targets. The results show that following their conjugation, both the effector and the target cells show significant reductions (>10%) in IFFP values. Changes in IFFP were induced by specific interaction and only between viable cells. No evidence of fluorescein transfer from a stained cell to its nonstained counterpart was found. To the best of our knowledge, this is the first time that effector-target interaction is monitored on an individual cell basis within a population, by means of IFFP measurements. In addition, in order to explain the physical phenomena, measurements of physical parameters which might affect the IFFP, such as changes in osmolality and pH, were performed and discussed. © 1998 Society of Photo-Optical Instrumentation Engineers.
本研究的目的是检测靶细胞和效应细胞在室温下结合后诱导的裂解前细胞内变化。使用Cellscan仪器通过细胞内荧光素荧光偏振(IFFP)来测量细胞质基质的变化。自然杀伤细胞和淋巴细胞激活杀伤细胞均用作效应细胞,而K562和Daudi细胞系用作靶细胞。结果表明,结合后,效应细胞和靶细胞的IFFP值均显著降低(>10%)。IFFP的变化是由特异性相互作用诱导的,且仅发生在活细胞之间。未发现荧光素从染色细胞转移至未染色细胞的证据。据我们所知,这是首次通过IFFP测量在群体内的单个细胞基础上监测效应细胞 - 靶细胞相互作用。此外,为了解释物理现象,还进行并讨论了可能影响IFFP的物理参数测量,如渗透压和pH值的变化。© 1998 光电仪器工程师协会。