Department of Chemistry, University of California Berkeley, Berkeley, California.
Department of Molecular & Cell Biology, University of California Berkeley, Berkeley, California.
Curr Protoc. 2021 Mar;1(3):e48. doi: 10.1002/cpz1.48.
Accurately mapping changes in cellular membrane potential across large groups of neurons is crucial for understanding the organization and maintenance of neural circuits. Measuring cellular voltage changes by optical means allows greater spatial resolution than traditional electrophysiology methods and is adaptable to high-throughput imaging experiments. VoltageFluors, a class of voltage-sensitive dyes, have recently been used to optically study the spontaneous activity of many neurons simultaneously in dissociated culture. VoltageFluors are particularly useful for experiments investigating differences in excitability and connectivity between neurons at different stages of development and in different disease models. The protocols in this article describe general procedures for preparing dissociated cultures, imaging spontaneous activity in dissociated cultures with VoltageFluors, and analyzing optical spontaneous activity data. © 2021 Wiley Periodicals LLC. Basic Protocol 1: Preparation of dissociated rat hippocampal or cortical cultures Alternate Protocol: Preparation of microisland dissociated cultures Basic Protocol 2: Imaging of spontaneous activity in dissociated cultures using voltage-sensitive dyes Basic Protocol 3: Analysis of spontaneous activity imaging data.
准确绘制大量神经元细胞膜电位变化图对于理解神经回路的组织和维持至关重要。通过光学手段测量细胞电压变化比传统的电生理学方法具有更高的空间分辨率，并且适用于高通量成像实验。电压荧光染料是一类电压敏感染料，最近已被用于在分离培养物中同时对许多神经元的自发活动进行光学研究。电压荧光染料对于研究不同发育阶段和不同疾病模型的神经元之间的兴奋性和连接性差异的实验特别有用。本文中的方案描述了制备分离培养物、用电压荧光染料对分离培养物中的自发活动进行成像以及分析光学自发活动数据的一般程序。© 2021 威立出版公司。基本方案 1：分离的大鼠海马或皮质培养物的制备可选方案：微岛分离培养物的制备基本方案 2：使用电压敏感染料对分离培养物中的自发活动进行成像基本方案 3：自发活动成像数据的分析。
