Bals S, Bechem M, Paffhausen W, Pott L
Institut für Zellphysiologie, Ruhr-Universität, Bochum, FRG.
Cell Calcium. 1990 Jun-Jul;11(6):385-96. doi: 10.1016/0143-4160(90)90050-5.
A setup for dual wavelength-excitation fluorescence measurements is introduced which permits a temporal resolution of up to 1 KHz, using the Ca2(+)-sensitive fluorescent dye Fura-2. The system makes use of a novel technical solution for chopping between two excitation wavelengths which does not move any optical components. Two beams, which are alternatively opened or shut by a rotating chopper wheel, are united by a dichroic mirror and are used for low-noise epifluorescence microscopy. The system includes a device for fast changes of extracellular solution that can be used for studying various components of [Ca2+]i-regulation in excitable and non-excitable cells. Sample recordings of spontaneous and experimentally-evoked [Ca2+]i-transients from cardiac myocytes are presented. Cardiac myocytes are a cell species that produces particularly fast [Ca2+]i-transients and therefore, a high temporal resolution is required in order to study physiological and/or pharmacological properties of these transients.
介绍了一种用于双波长激发荧光测量的装置,该装置使用对Ca2(+)敏感的荧光染料Fura-2,可实现高达1千赫兹的时间分辨率。该系统采用了一种新颖的技术解决方案,用于在两个激发波长之间进行切换,无需移动任何光学组件。由旋转斩光轮交替打开或关闭的两束光,通过二向色镜合并,用于低噪声落射荧光显微镜。该系统包括一个用于快速改变细胞外溶液的装置,可用于研究可兴奋细胞和不可兴奋细胞中[Ca2+]i调节的各种成分。展示了来自心肌细胞的自发和实验诱发的[Ca2+]i瞬变的样本记录。心肌细胞是一种产生特别快速的[Ca2+]i瞬变的细胞类型,因此,为了研究这些瞬变的生理和/或药理特性,需要高时间分辨率。