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Thomas Cover Glass Staining Outfits

托马斯盖玻璃染色服装

Company: Thomas Scientific
Catalog#: 8542E40
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Cloud-point PEG Glass Surfaces for Imaging of Immobilized Single Molecules by Total-internal-reflection Microscopy
Author:
Date:
2016-04-05
[Abstract]  This effective, robust protocol generates glass coverslips coated with biotin-functionalized polyethylene glycol (PEG), making the glass surface resistant to non-specific absorption of biomolecules, and permitting immobilization of biomolecules for subsequent single-molecule tracking of biochemical reactions. The protocol can be completed in one day, and the coverslips can be stored for at least 1 month. We have confirmed that the PEG surfaces prepared according to the protocol are resistant to non-specific adsorption by a wide range of biomolecules (bacterial, mitochondrial, and human transcription factors, DNA, and RNA) and biological buffers. [摘要]  这种有效的,稳健的方案产生涂覆有生物素官能化聚乙二醇(PEG)的玻璃盖玻片,使得玻璃表面对生物分子的非特异性吸收具有抗性,并允许固定生物分子以用于随后的单分子跟踪生化反应。 协议可以在一天内完成,盖玻片可以存储至少1个月。 我们已经证实,根据方案制备的PEG表面可以通过广泛的生物分子(细菌,线粒体和人转录因子,DNA和RNA)和生物缓冲液的非特异性吸附抗性。
【背景】研究生物分子动力学的单分子成像方法通过允许实时跟踪多步反应而不需要同步试剂来补充传统的“大量”生物化学方法(Weiss,1999)。在大多数单分子成像方法中,首先用单个荧光团标记感兴趣的生物分子,然后将标记的生物分子固定在光学透明的表面(通常为玻璃或二氧化硅)上,并且被检测为衍射受限图像(“斑点”)使用配有高灵敏度相机的光学显微镜(Selvin和Ha,2008)。表面固定有两个目的。首先,它允许在超过几百毫秒的时间尺度上跟踪分子态(否则,生物分子将扩散到焦平面外)。第二,表面允许在全内反射几何中激发荧光(Axelrod,1981),其显着地增加了与表面接近(<100nm)位置的分子的检测的信噪比(selvin和哈,2008)。尽管有这些明显的优势,表面也是单分子分析中最常见的人为因素(visnapuu et=""> ...

Neuron Culture from Mouse Superior Cervical Ganglion
Author:
Date:
2014-01-20
[Abstract]  The rodent superior cervical ganglion (SCG) is a useful and readily accessible source of neurons for studying the mechanisms of sympathetic nervous system (SNS) development and growth in vitro. The sympathetic nervous system (SNS) of early postnatal animals undergoes a great deal of remodeling and development; thus, neurons taken from mice at this age are primed to re-grow and establish synaptic connections after in situ removal. The stereotypic location and size of the SCG make it ideal for rapid isolation and dissociation. The protocol described here details the requirements for the dissection, culture and differentiation of SCG neurons. The protocol is suitable for culturing neurons from late embryonic gestation to approximately postnatal day 3. The culture technique ... [摘要]  啮齿动物上级子宫颈神经节(SCG)是一种有用且易于获取的神经元来源,用于研究交感神经系统(SNS)体外发育和生长的机制。 早期出生后动物的交感神经系统(SNS)经历了大量的重塑和发育; 因此,在这个年龄的小鼠中取出的神经元被引发再生长并在原位移除后建立突触连接。 SCG的刻板位置和尺寸使其成为快速隔离和解离的理想选择。 这里描述的方案详述了SCG神经元的解剖,培养和分化的要求。 该方案适用于从晚期胚胎妊娠至大约出生后第3天培养神经元。下面讨论的培养技术利用玻璃盖玻片进行固定细胞的显微镜检查。

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