The resulting ligand-lipid vesicle solution was translucent. Then it was incubated in the dark for 12h before use. Transmission electron microscopy (TEM) was carried out on a JEM-100CX II electronmicroscope. The sample was prepared by freeze-fracture replication according to standard techniques with a high-vacuum freeze-etching system(Balzers BAF-400D). freeze-fracture replication /high-vacuum freeze-etching 是什么意思啊？是不是得到的溶液是透明或半透明时必须通过这种技术才能用TEM观察，染色是不是也需冻结才有效？ 我刚刚看几篇文献都是这样做的，对这方面不怎么了解，望大家多多给予指教
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