活细胞成像可以通过监测钙通量来揭示动态和多模态细胞信号传导。Ca2+浓度的时空变化激发了特定的下游过程,并通过对这些事件进行分类,我们可以检查语言细胞用来与自己和彼此交流。因此,钙成像是一种可以理解的流行和通用的技术,它依赖于通过荧光强度测量的高分辨率光学数据.这在贴壁细胞上相对容易地执行,作为荧光强度的变化可以随时间在感兴趣的固定区域中监测。然而,非粘附或轻度粘附的细胞的灌注导致它们的机械位移,从而阻碍荧光强度随时间变化的空间分辨率。在这里,我们提供了使用明胶防止在记录期间发生的溶液交换期间细胞移位的简单且具有成本效益的方案的细节。
Live-cell imaging can reveal dynamic and multimodal cell signaling by monitoring calcium flux. Spatiotemporal changes in Ca2+ concentrations instigate specific downstream processes and by categorizing these events, we can examine the language cells use to communicate both to themselves and with each other. Thus, calcium imaging is an understandably popular and versatile technique that relies on high-resolution optical data as measured by fluorescence intensity. This is executed with relative ease on adherent cells, as changes in fluorescence intensity can be monitored over time in fixed regions of interest. However, perfusion of non-adherent or mildly adherent cells leads to their mechanical displacement thereby hindering the spatial resolution of fluorescence intensity changes through time. Here we provide details of a simple and cost-effective protocol using gelatin to prevent cell dislodgement during the solution exchanges that occur during recording.