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Sucrose

蔗糖

Company: Sigma-Aldrich
Catalog#: S8501
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Registration and Alignment Between in vivo Functional and Cytoarchitectonic Maps of Mouse Visual Cortex
Author:
Date:
2018-02-20
[Abstract]  This protocol describes a method for registration of in vivo cortical retinotopic map with cytochrome c oxidase (CO) labeled architectonic maps of the same mouse brain through the alignment of vascular fiducials. By recording surface blood vessel pattern and sequential alignment at each step, this method overcomes the challenge imposed by tissue distortion during perfusion, mounting, sectioning and histology procedures. This method can also be generalized to register and align other types of in vivo functional maps like ocular dominance map and spatial/temporal frequency tuning map with various anatomical maps of mouse cortex. [摘要]  该协议描述了通过血管基准点的对齐使用细胞色素c氧化酶(CO)标记的相同小鼠脑的构建图来注册体内皮质视网膜地图的方法。 通过记录每个步骤的表面血管图案和顺序对准,该方法克服了在灌注,贴壁,切片和组织学程序期间由组织变形所施加的挑战。 这种方法也可以推广到注册和对齐其他类型的体内功能地图,如眼优势地图和空间/时间频率调整地图与小鼠皮层的各种解剖图。

【背景】通过体内视网膜映射(Marshel等人,2011; Garrett等人,),可以将小鼠视觉皮层分隔成功能上不同的视觉区域。 (Olavarria and Montero,1989; Wang and Burkhalter,2007),或者通过建筑结构辅助的神经元追踪技术,这些不同的视觉区域具有不同的响应特性和皮质皮质连接(Andermann等人,2011; Marshel等人,2011; Roth等人, >,2012; Wang等人,2011和2012)。这些结果表明,鼠标视觉区域形成处理不同类型的视觉信息的分离的视觉流(Murakami等人,2017; Smith等人,2017)。在视觉区域地图的背景下研究鼠标视觉系统对于理解视觉皮层的组织是至关重要的。然而,虽然功能图和结构图大致相似,但是这两幅图显示的并不完美(Zhuang et ...

Visualising Differential Growth of Arabidopsis Epidermal Pavement Cells Using Thin Plate Spline Analysis
Author:
Date:
2016-11-20
[Abstract]  Epidermal pavement cells in Arabidopsis leaves and cotyledons develop from relatively simple shapes to form complex cells that have multiple undulations of varying sizes. Analyzing the growth of individual parts of the cell wall boundaries over time is essential to understanding how pavement cells develop their complex shapes. Thin plate spline analysis is a method for visualizing the change of size and shape of objects through warping or deformation of a regular mesh and can be applied to understand cell wall growth. This protocol describes the application of thin plate spline analysis to visualize the development of individual pavement cells over time. [摘要]  拟南芥叶和子叶中的表皮铺路细胞从相对简单的形状发育而形成具有不同大小的多个起伏的复合细胞。分析细胞壁边界的各个部分随时间的生长对理解铺路细胞如何发展其复杂形状是至关重要的。薄板样条分析是通过规则网格的翘曲或变形来可视化物体的尺寸和形状的变化的方法,并且可以用于理解细胞壁生长。该协议描述了薄板样条分析的应用,以便随时间可视化单个路面细胞的发育。

[背景] 了解细胞生长的空间模式提供了洞察植物细胞如何形成不同的形状。拟南芥子叶和叶的表皮铺路细胞是用于研究复杂细胞如何生长的良好模型系统,因为它们的细胞壁边界从最初为简单弧的边界开始形成不同大小的多个起伏(Armor et al。,2015; Fu et al。,2005)。通过将外部施加的标记物固定到细胞例如藻类氮细胞节间(Green等人,1970),根细胞(Shaw等人, ,2000)和毛状体(Schwab等人,2003)。然而,从外部施加的界标测量细胞生长有时是不可行的,例如当外部施加的荧光标记物的强荧光会遮蔽细胞内荧光标记的细胞骨架元件时(Armor等人,2015)。显示定义数量的同源界标随时间或在不同物体之间的变化位置的薄板样条分析先前已用于分析诸如人类头骨的物体的三维大小和形状的变化(Rosas和Bastir,2002; Gunz ,2014)和叶子(Polder et al ...

Lipid Extraction from HeLa Cells, Quantification of Lipids, Formation of Large Unilamellar Vesicles (LUVs) by Extrusion and in vitro Protein-lipid Binding Assays, Analysis of the Incubation Product by Transmission Electron Microscopy (TEM) and by Flotation across a Discontinuous Sucrose Gradient
Author:
Date:
2016-10-20
[Abstract]  Dissecting the interactions established between proteins and membranes in a given type of cells is not an easy task. Using a cell-free system of large unilamellar vesicles (LUVs) to analyze these interactions may help decipher these interactions and identify potential membrane deformations induced by the proteins incubated with these LUVs. This article describes the protocols for 1) extraction of total lipids from eukaryotic cells using the method developed by Bligh and Dyer (1959), 2) the quantification of glycerophospholipids by gas chromatography after methanolysis, followed by 3) the formation of LUVs by extrusion, 4) protein-lipid binding assay, 5) analysis of the incubation product by transmission electron microscopy (TEM) and by flotation across a discontinuous sucrose gradient and ... [摘要]  解剖在给定类型的细胞中蛋白质和膜之间建立的相互作用不是一个容易的任务。使用大单层囊泡(LUV)的无细胞系统来分析这些相互作用可以帮助破译这些相互作用和识别由与这些LUV孵育的蛋白质诱导的潜在的膜变形。本文介绍了1)使用由Bligh和Dyer(1959)开发的方法从真核细胞中提取总脂质,2)在甲醇分解后通过气相色谱法定量甘油磷脂,然后3)通过挤出形成LUV的方案, 4)蛋白质 - 脂质结合测定,5)通过透射电子显微镜(TEM)和通过不连续蔗糖梯度浮选分析孵育产物,最后,6)通过免疫印迹分析蛋白质并通过碘素熏蒸显示甘油磷脂。

[背景] 包含巨单层囊泡(GUV;由单个磷脂双层组成,直径大于1μm)或脂质体孵育的无细胞系统与重组蛋白可能有助于了解这些相互作用。根据它们的直径和层数,脂质体被分为小的单层囊泡(SUV;由单个磷脂双层构成的囊泡,直径在20和100nm之间),大的单层囊泡(LUV;由单个双层磷脂,并且直径在100和400nm之间),大多层囊泡(MLV;由多个磷脂双层构成且直径在200nm和3μm之间的囊泡)和多泡囊泡(MVV);由囊泡组成的大囊泡单个双层磷脂,并含有几个较小的囊泡,每个囊泡由单个双层磷脂组成)。 ...

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