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  • Protocol for BATTLE-1EX: A High-Resolution Imaging Method to Visualize Whole Synaptic Structures and their Components in the Nervous System.

Protocol for BATTLE-1EX: A High-Resolution Imaging Method to Visualize Whole Synaptic Structures and their Components in the Nervous System.

STAR protocols (2020-12-31)
Akitoshi Inoue, Takuya Kobayashi, Hirokazu Hirai, Noriko Kanaya, Keigo Kohara
摘要

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).

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Sigma-Aldrich
杜氏改良 Eagle 培养基 - 高葡萄糖, With 4500 mg/L glucose, sodium pyruvate, and sodium bicarbonate, without L-glutamine, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
过硫酸铵, Molecular Biology, suitable for electrophoresis, ≥98%
Sigma-Aldrich
四甲基乙二胺, BioReagent, Molecular Biology, ≥99% (GC)
Sigma-Aldrich
丙烯酸钠, 97%
Sigma-Aldrich
丙烯酰胺, Molecular Biology, ≥99% (HPLC)
Sigma-Aldrich
N,N′-亚甲基双丙烯酰胺, powder, Molecular Biology, suitable for electrophoresis, ≥99.5%
Sigma-Aldrich
哌啶醇氧化物, 95%