| FM1-43 Photoconversion and Electron Microscopy Analysis at the Drosophila Neuromuscular Junction
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Author:
Date:
2017-09-05
[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; ...
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| Determination of Molecular Structures of Condensed Tannins from Plant Tissues Using HPLC-UV Combined with Thiolysis and MALDI-TOF Mass Spectrometry
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Author:
Date:
2016-10-20
[Abstract] Condensed tannins extracted from plant tissues are suitable substitutes for phenolic resins. Their molecular structure, which might influence their chemical reactivity, can be assessed by the use of both HPLC-UV after acid thiolysis and MALDI-TOF mass spectrometry. Thiolysis of plant extracts in acidic methanol with cysteamine hydrochloride results in the release of the monomeric units of the condensed tannin oligomers that can be further quantified by reversed-phase HPLC-UV by comparison with analytical standards. MALDI-TOF mass spectrometry using 2,5-dihydroxybenzoic acid as matrix and K+ as cationization agent highlights the molecular structural characteristics (e.g., monomeric unit sequence) of the tannin oligomers. The methodologies permit the estimation of the ...
[摘要] 从植物组织提取的缩合单宁是酚醛树脂的合适替代物。它们的可能影响它们的化学反应性的分子结构可以通过在酸硫解和MALDI-TOF质谱之后使用HPLC-UV来评估。用半胱胺盐酸盐在酸性甲醇中溶解植物提取物导致缩合的单宁寡聚体的单体单元的释放，其可以通过与分析标准比较通过反相HPLC-UV进一步定量。使用2,5-二羟基苯甲酸作为基质和K sup +作为阳离子化试剂的MALDI-TOF质谱分析突出了单宁的分子结构特征（例如单体单元序列）低聚物。该方法允许估计平均和最大（可观察）聚合度，单体单元的类型和单宁单体的糖基化和/或酯化的存在。
[背景] 缩合单宁是由可从几种植物组织（例如软木树皮）中提取的黄烷-3-醇单体单元组成的多酚低聚物。它们已被认为是树脂配方（例如木材粘合剂和泡沫材料）中合成酚醛树脂的合适替代品。在缩合鞣酸中检测到的最常见的黄烷-3-醇单体，其羟基化模式和立体化学不同，如图1所示。
![]() "src ="/attached/image/20161012/20161012024340_4416.jpg"/>
图1.在缩合单宁结构中确定的最常见的单体
低聚物中单体单元的具体结构和聚合度强烈影响单宁的化学反应性和物理性质，例如与醛的缩合反应速率，重金属螯合能力和水溶液的粘度溶液（Pizzi和Stephanou，1994; ...
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| Micro Neutralization (MN) Assay of Influenza Viruses with Monoclonal Antibodies
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Author:
Ying Wu, MyungSam Cho, David Shore, Manki Song, JungAh Choi, Tao Jiang, Yong-Qiang Deng, Melissa Bourgeois, Lynn Almli, Hua Yang, Li-Mei Chen, Yi Shi, Jianxu Qi, An Li, Kye Sook Yi, MinSeok Chang, Jin Soo Bae, HyunJoo Lee, JiYoung Shin, James Stevens, SeoungSuh Hong, Cheng-Feng Qin, George F. Gao, Shin Jae Chang and Ruben O. Donis,
Date:
2016-06-05
[Abstract] The human monoclonal antibodies generated from single human B cells were tested to characterize their ability to neutralize virus infectivity. The microneutralization assay is a highly sensitive and specific assay for detecting virus-specific neutralizing antibodies to influenza viruses. This protocol is to measure the ability of human monoclonal antibody to neutralize influenza virus by microneutralization assay.
[摘要] 测试从单个人B细胞产生的人单克隆抗体，以表征其中和病毒感染性的能力。 微量中和测定法是用于检测流感病毒的病毒特异性中和抗体的高度灵敏和特异性的测定法。 该方案是测量人单克隆抗体通过微量中和测定中和流感病毒的能力。
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