Sabeva Nadezhda S, Bykhovskaia Maria
Neuroscience Department, Universidad Central del Caribe, Bayamón, Puerto Rico, USA.
Department of Neurology, Department of Anatomy and Cell Biology, Wayne State University School of Medicine, Detroit, Michigan, USA.
Bio Protoc. 2017 Sep 5;7(17). doi: 10.21769/BioProtoc.2523.
We developed a protocol for photoconversion of endocytic marker FM1-43 followed by electron microscopy analysis of synaptic boutons at the neuromuscular junction. This protocol allows detection of stained synaptic vesicle even when release rates are very low, such as during the spontaneous release mode. The preparations are loaded with the FM1-43 dye, pre-fixed, treated and illuminated to photoconvert the dye, and then processed for conventional electron microscopy. This procedure enables clear identification of stained synaptic vesicles at electron micrographs.
我们开发了一种用于内吞标记物FM1-43光转化的方案，随后对神经肌肉接头处的突触小体进行电子显微镜分析。即使在释放速率非常低的情况下，如在自发释放模式期间，该方案也能检测到染色的突触小泡。将制剂用FM1-43染料加载、预固定、处理并照射以光转化染料，然后进行常规电子显微镜处理。该程序能够在电子显微镜照片上清晰地识别染色的突触小泡。
