Department of Medical Chemistry, Kansai Medical University, Graduate School of Medicine, Hirakata, Osaka 573-1010, Japan.
Department of Neurophysiology and Neural Repair, Gunma University Graduate School of Medicine, Maebashi, Gunma 371-8511, Japan.
STAR Protoc. 2020 Nov 25;1(3):100166. doi: 10.1016/j.xpro.2020.100166. eCollection 2020 Dec 18.
This protocol describes BATTLE-1EX, which is a combined method of BATTLE-1 and expansion microscopy to obtain high-resolution imaging of whole synaptic structures and their components of hippocampal neural circuits. BATTLE-1 uses two genetically engineered recombinase proteins and competition between two recombinases that can be independently titrated, resulting in a tunable proportion of mCherry+/YFP- and YFP+/mCherry- cells. As a combinational method, BATTLE-1EX has the potential to visualize and dissect whole synaptic structures in numerous regions in the brain. For complete details on the use and execution of this protocol, please refer to Kohara et al. (2020).
本方案描述了 BATTLE-1EX,这是一种结合 BATTLE-1 和扩展显微镜的方法,可获得海马神经回路全突触结构及其成分的高分辨率图像。BATTLE-1 使用两种基因工程重组酶蛋白和两种可独立滴定的重组酶之间的竞争,从而产生 mCherry+/YFP-和 YFP+/mCherry-细胞的可调比例。作为一种组合方法,BATTLE-1EX 有可能可视化和剖析大脑中许多区域的全突触结构。有关此方案使用和执行的完整详细信息,请参见 Kohara 等人。(2020)。