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Sodium Phosphate Monobasic

磷酸氢二钠

Company: Wako Pure Chemical Industries
Catalog#: 197-02865
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A Streamlined Method for the Preparation of Gelatin Embedded Brains and Simplified Organization of Sections for Serial Reconstructions
Author:
Date:
2017-11-20
[Abstract]  Gelatin embedding of whole brains for sectioning is a critical procedure used in neuroscience to ensure all morphological and spatial details are preserved intact. Here, we describe an inexpensive, reproducible and efficient means to embed post-fixed brains ready for sectioning in gelatin within a week’s time. The sections obtained are distortion-free and their fragile internal structures preserved which can be used for serial reconstructions for lesion studies and mapping of viral expression after stereotaxic injections. In addition, the separation of adjacent slices into a series of 3-4 vials facilitates subsequent organization and assembly of serial sections at the mounting step. [摘要]  整个脑切片明胶嵌入是神经科学中使用的关键程序,以确保所有的形态和空间的细节保存完好。 在这里,我们描述了一个廉价,可重复和有效的方法来嵌入后固定的大脑准备切片明胶一个星期的时间。 获得的部分是无畸变的,它们的脆弱内部结构被保留下来,可用于系列重建病变研究和立体定位注射后的病毒表达图谱。 此外,将相邻切片分离成3-4个小瓶系列便于在安装步骤中对连续切片进行组织和组装。
【背景】行为神经科学的最新进展已经允许将视蛋白和抗体靶向毒素引入特定亚群的神经元和大脑区域。这些研究通常需要全脑切片的可视化用于组织学和形态学分析,以通过用于行为验证的立体定位注射(Aoki等,,2015)或引言定位细胞类型特异性抗体靶向毒素诱导的损伤的病毒递送转基因(Aquili等人,2014)。利用狂犬病病毒(Suzuki等人,2012)对神经元回路进行了详细的绘图,并且使用明胶作为包埋剂实现了连续切片的定位调查,所述包埋剂作为组织内和周围的结构基质。明胶浸渍的脑组织提供了加强的支持精致的内部结构,如海马和脑室空间,这是容易损坏免疫组织化学(IHC)处理和随后安装到幻灯片上。嵌入式脑切片在安装时通常也不会变形,并且可以将相邻切片用于连续重建。

凝胶嵌入和IHC处理后的质量取决于几个程序性的细节,这可能是乏味和耗时的。在先前公开的方案中,明胶向脑和心室空间的适当渗透和渗透需要真空烘箱(Griffioen等,1992)。此外,将大脑嵌入小模具中是具有挑战性的,因为大脑具有漂浮的趋势。另外,在连续重构的IHC处理之后识别和定向相邻切片可能是艰巨的。 ...

EAE Induction by Passive Transfer of MOG-specific CD4+ T Cells
Author:
Date:
2017-07-05
[Abstract]  Experimental autoimmune encephalomyelitis (EAE) is an animal model of multiple sclerosis (MS), which is a chronic inflammatory disease of the central nervous system (CNS). It is characterized by focal demyelination and inflammatory responses mediated by myelin-specific autoreactive CD4+ T cells. Using a passive transfer model of EAE in mice, we have demonstrated that regional specific neural signals by sensory-sympathetic communications create gateways for immune cells at specific blood vessels of the CNS, a phenomenon known as the gateway reflex (Arima et al., 2012; Tracey, 2012; Arima et al., 2013; Sabharwal et al., 2014; Arima et al., 2015b). Here we describe protocols for passive transfer model of EAE using freshly isolated (MOG)-specific ... [摘要]  实验性自身免疫性脑脊髓炎(EAE)是多发性硬化症(MS)的动物模型,其是中枢神经系统(CNS)的慢性炎性疾病。其特征在于由髓磷脂特异性自身反应性CD4 + T细胞介导的局灶性脱髓鞘和炎症反应。在小鼠中使用EAE的被动转移模型,我们已经证明,通过感觉交感通信的区域特异性神经信号在CNS的特定血管上产生免疫细胞的网关,这被称为网关反射(Arima等, ,2012; Tracey,2012; Arima等人,2013; Sabharwal等人,2014; Arima等人。 >,2015b)。在这里,我们描述了使用新鲜分离的(MOG)特异性CD4 + T细胞或周期性再刺激的MOG特异性CD4 + T细胞系的EAE的被动转移模型的方案,其是适用于体内追踪致病性CD4 T细胞,特别是CNS(Ogura等人,2008; Arima等人) 。,2012和2015b)。
【背景】广泛接受的是,自身反应性CD4 + T细胞在MS和EAE的发病机理中起重要作用(Reboldi,2009; International Multiple Sclerosis Genetics,et al。,2011; Steinman,2014),它们是CNS的慢性炎性疾病。 ...

In vivo Live Imaging of Calcium Waves and Other Cellular Processes during Fertilization in Caenorhabditis elegans
Author:
Date:
2017-04-05
[Abstract]  Fertilization calcium waves are a conserved trigger for animal development; however, genetic analysis of these waves has been limited due to the difficulty of imaging in vivo fertilization. Here we describe a protocol to image calcium dynamics during in vivo fertilization in the genetic animal model Caenorhabditis elegans. This protocol consists of germline microinjection of a chemical calcium indicator, worm immobilization, live imaging, and image processing that quantifies the calcium fluorescence in the oocyte region moving in the field-of-view during ovulation. This imaging protocol can also be used to image other cellular processes during in vivo fertilization in C. elegans, such as membrane fusion and cytoskeletal dynamics. [摘要]  受精钙波是动物发育的保守触发因素; 然而,这些波的遗传分析由于在体内成像难以受精而受到限制。 在这里,我们描述了在遗传动物模型秀丽隐杆线虫中的体内受精期间成像钙动力学的方案。 该方案包括化学钙指示剂的种系显微注射,蠕虫固定,活体成像和图像处理,其量化在排卵期间在视野内移动的卵母细胞区域中的钙荧光。 该成像方案也可以用于在体内受精期间对其他细胞过程进行成像。 线虫,如膜融合和细胞骨架动力学。

受精钙波在卵活化中起关键作用,并通过使用体外施肥系统在各种生物体中进行了分析。 线虫秀丽隐杆线虫由于其半透明的身体,适合于在体内成像受精。 受精钙成像在C. 塞缪尔等人报道了使用化学钙指示剂的线虫。 (2001)。 我们在这里描述了通过应用高速旋转盘共焦显微镜和图像处理方法从成像方法修改的方案,其在排卵期间分割在视场中移动的受精卵母细胞区域。 该协议能够精确定量描述钙波的时间动力学和波形图的遗传分析。

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