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Potassium chloride

氯化钾

Company: Sigma-Aldrich
Catalog#: P3911
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Structural Analysis of Bordetella pertussis Biofilms by Confocal Laser Scanning Microscopy
Author:
Date:
2018-08-05
[Abstract]  Biofilms are sessile communities of microbial cells embedded in a self-produced or host-derived exopolymeric matrix. Biofilms can both be beneficial or detrimental depending on the surface. Compared to their planktonic counterparts, biofilm cells display enhanced resistance to killing by environmental threats, chemicals, antimicrobials and host immune defenses. When in biofilms, the microbial cells interact with each other and with the surface to develop architecturally complex multi-dimensional structures. Numerous imaging techniques and tools are currently available for architectural analyses of biofilm communities. This allows examination of biofilm development through acquisition of three-dimensional images that can render structural features of the sessile community. A frequently ... [摘要]  生物膜是嵌入自生或宿主衍生的外聚合物基质中的微生物细胞的固着群落。根据表面,生物膜可以是有益的或有害的。与浮游生物相比,生物膜细胞表现出更强的抗环境威胁,化学物质,抗菌药物和宿主免疫防御能力。当处于生物膜中时,微生物细胞彼此相互作用并与表面相互作用以形成结构复杂的多维结构。目前,许多成像技术和工具可用于生物膜群落的建筑分析。这允许通过获取可以呈现无柄群落的结构特征的三维图像来检查生物膜的发展。经常使用的工具是共聚焦激光扫描显微镜。我们提出了一个详细的协议,以生长,观察和分析呼吸道人类病原体,百日咳博德特氏菌的生物膜在空间和时间。

【背景】百日咳博德特氏菌(Bordetella pertussis)是上呼吸道的专性人类病原体,引起百日咳或百日咳(Mooi,2010; Dorji et al。,2018)。 B的生物膜。百日咳在各种人造表面上以及静态,摇动和流体流动条件下形成(Mishra et al。,2005; Sloan et al。,2007 ; Serra et al。,2011)。对这些生物膜的显微评估表明,这种细菌产生不规则形状的微集落,由流体通道分隔,嵌入由细胞外DNA(eDNA),蛋白质和多糖组成的外聚合物基质中(Parise et al。,2007; Sloan et al。,2007; Serra et al。,2008; ...

Dual-sided Voltage-sensitive Dye Imaging of Leech Ganglia
Author:
Date:
2018-03-05
[Abstract]  In this protocol, we introduce an effective method for voltage-sensitive dye (VSD) loading and imaging of leech ganglia as used in Tomina and Wagenaar (2017). Dissection and dye loading procedures are the most critical steps toward successful whole-ganglion VSD imaging. The former entails the removal of the sheath that covers neurons in the segmental ganglion of the leech, which is required for successful dye loading. The latter entails gently flowing a new generation VSD, VF2.1(OMe).H, onto both sides of the ganglion simultaneously using a pair of peristaltic pumps. We expect the described techniques to translate broadly to wide-field VSD imaging in other thin and relatively transparent nervous systems. [摘要]  在这个协议中,我们介绍了一种有效的方法,用于Tomina和Wagenaar(2017)中使用的电压敏感染料(VSD)加载和水蛭神经节成像。 解剖和染料加载程序是成功完成全神经节VSD成像的关键步骤。 前者需要去除覆盖水蛭节段神经节神经元的鞘,这是成功染料加载所需的。 后者需要使用一对蠕动泵同时轻柔地将新一代VSD VF2.1(OMe).H流入神经节的两侧。 我们期望所描述的技术广泛地转化为其他薄且相对透明的神经系统中的宽视场VSD成像。

【背景】双面显微镜是一种宽视野荧光成像系统,由一对精确对准的显微镜组成,用于观察来自对面的神经元制剂并且一次显示不同的焦平面(Tomina and Wagenaar,2017)。通过将该光学系统与新一代电压敏感染料(VSD),VoltageFluor(Miller等人,2012; Woodford等人,2015),荧光可以同时从不同深度的神经元捕获编码具有高保真度膜电压的信号。我们将这种泛神经元记录系统应用于药用水蛭的神经系统,我们利用电生理学方法诱发虚构行为并定量控制可识别神经元的膜电位(Tomina and ...

High Throughput NPY-Venus and Serotonin Secretion Assays for Regulated Exocytosis in Neuroendocrine Cells
Author:
Date:
2018-01-05
[Abstract]  Here we describe two assays to measure dense core vesicle (DCV) exocytosis-mediated cargo secretion in neuroendocrine cells. To conduct siRNA screens for novel genes in regulated DCV exocytosis, we developed a plate reader-based secretion assay using DCV cargo, NPY-Venus, and an orthogonal 3H-serotonin secretion assay. The NPY-Venus secretion assay was successfully used for a high throughput siRNA screen, and the serotonin secretion assay was used to validate hits identified from the screen (Sorensen, 2017; Zhang et al., 2017). [摘要]  在这里我们描述了两种测定方法来测量神经内分泌细胞中的致密核心囊泡(DCV)胞吐作用介导的货物分泌。 为了在受调控的DCV胞吐作用中对新基因进行siRNA筛选,我们开发了使用DCV货物,NPY-金星以及正交3H-血清素分泌测定的基于读板器的分泌测定法。 NPY-金星分泌测定成功地用于高通量siRNA筛选,并且使用血清素分泌测定来验证从筛选中鉴定的命中(Sorensen,2017; Zhang等人,2017)。


【背景】致密核心囊泡(DCV)胞吐作用介导内分泌和神经内分泌细胞分泌蛋白质,肽和小分子。当蛋白质和肽激素被内质网上的核糖体合成时,蛋白质和肽激素进入分泌途径,并在跨高尔基网络(TGN)(Borgonovo等人,2006; Bowman等, >等人,2009)。小分子物质,如5-羟色胺,直接被胞质单胺转运蛋白(VMATs)从细胞质直接吸收到DCV的内腔(Sudhof,2004)。在非刺激条件下,货物分子被储存在靠近质膜的DCV内。当细胞接受增加细胞质Ca ...

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