Oviedo Néstor J, Nicolas Cindy L, Adams Dany S, Levin Michael
Center for Regenerative and Developmental Biology, Forsyth Institute and Developmental Biology Department, Harvard School of Dental Medicine, Boston, MA 02115, USA.
CSH Protoc. 2008 Oct 1;2008:pdb.prot5055. doi: 10.1101/pdb.prot5055.
INTRODUCTIONThis protocol describes how to use the anionic membrane voltage-reporting dye DiBAC(4)(3) to generate images of cell membrane potential in live planarians. These images qualitatively reveal variations in time-averaged membrane potential across different regions of the organism. Changes in these images due to experimental treatments reveal how the particular treatment affects this physiological parameter. This method is a great improvement over standard electrophysiological techniques, which cannot be used to gain an understanding of the electrical properties of an entire worm or a regenerating fragment, due to small cell size and large cell number. When the proper controls are performed, this technique is a very powerful and simple way to gather physiologic data.
引言
本方案描述了如何使用阴离子膜电压报告染料DiBAC(4)(3)来生成活涡虫细胞膜电位的图像。这些图像定性地揭示了整个生物体不同区域的时间平均膜电位变化。实验处理导致的这些图像变化揭示了特定处理如何影响这一生理参数。与标准电生理技术相比，该方法有很大改进，由于细胞体积小且数量多，标准电生理技术无法用于了解整个蠕虫或再生片段的电特性。当进行适当的对照实验时，该技术是一种收集生理数据的非常强大且简单的方法。
