{{'Search' | translate}}



Company: Leica Biosystems
Catalog#: Leica EM UC6
Other protocol()

FM1-43 Photoconversion and Electron Microscopy Analysis at the Drosophila Neuromuscular Junction
[Abstract]  We developed a protocol for photoconversion of endocytic marker FM1-43 followed by electron microscopy analysis of synaptic boutons at the Drosophila 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. [摘要]  我们开发了内吞标记FM1-43的光转换方案,然后在果蝇神经肌肉接头处进行突触引物的电子显微镜分析。 即使在释放速率非常低时,例如在自发释放模式期间,该方案允许检测染色的突触小泡。 该制剂装载有FM1-43染料,经预先固定,处理和照射,以使染料转变为染料,然后进行常规电子显微镜处理。 该方法能够在电子显微照片下清楚鉴定染色的突触小泡。
【背景】神经元发射体通过突触小泡与神经元质膜的融合而释放。囊泡可以自发融合或响应动作电位。随后,囊泡通过内吞作用获得回收。通过分子生物学,电生理学和显微镜的工具广泛研究了突触小泡回收的分子机制(Slepnev和De Camilli,2000; Sudhof,2004; Rizzoli和Betz,2005; Kavalali,2006)。加载内参标记FM1-43与染料光转换耦合,然后进行电子显微镜分析是一种强大的技术,允许调查和测量回收囊泡池(Harata et al。,2001; Schikorski and Stevens,2001; Rizzoli和Betz, 2004)。果蝇神经肌肉接头(NMJ)是具有明确定义的突触引物的有利制剂,其能够快速产生具有突变突触蛋白的细胞系和严格评估囊泡回收池(Akbergenova和Bykhovskaia,2009; ...

Cytohistochemical Determination of Calcium Deposition in Plant Cells
[Abstract]  Calcium plays important roles in maintaining plant cellular structure and also acts as a key secondary messenger in intercellular signaling. Thirty years ago, methods of detecting calcium in sub-cellular level had been established (Stockwell and Hanchey, 1982; Borgers et al., 1982) and reviewed extensively (Wick and Heplerm, 1982). We had used the method of testing calcium localization in salt tolerance improved transgenic alfalfa plant (Zhang and Wang, 2015). Here, we describe the protocol of testing calcium deposition by staining with potassium pyroantimonate (PPA) in detail, which was adapted from former reports (Stockwell and Hanchey, 1982; Borgers et al., 1982). The principle of this protocol is that the Ca2+ can react with antimonite and from black ... [摘要]  钙在维持植物细胞结构中起重要作用,并且还在细胞间信号传导中作为关键的第二信使。三十年前,已经建立了在亚细胞水平检测钙的方法(Stockwell和Hanchey,1982; Borgers等人,1982)并广泛地综述(Wick和Heplerm,1982)。我们使用测试钙定位的方法在耐盐改良的转基因苜蓿植物中(Zhang和Wang,2015)。在这里,我们描述了通过用详细的焦锑酸钾(PPA)染色测试钙沉积的方案,其改编自以前的报道(Stockwell和Hanchey,1982; Borgers等人,1982)。该方案的原理是Ca 2+ 2+可以与锑矿和黑色颗粒反应,这可以在透射电子显微镜下观察到。该协议包括植物组织的常见显微操作技术,用透射电子显微镜和照相观察。