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Company: Gibco
Catalog#: 14190250
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Non-separate Mouse Sclerochoroid/RPE/Retina Staining and Whole Mount for the Integral Observation of Subretinal Layer
Author:
Date:
2021-01-05
[Abstract]  

The subretinal layer between retinal pigment epithelium (RPE) and photoreceptors is a region involved in inflammation and angiogenesis during the procession of diseases such as age-related macular degeneration. The current protocols of whole mounts (retina and RPE) are unable to address the intact view of the subretinal layer because the separation between retina and RPE is required, and each separate tissue is then stained. Non-separate Sclerochoroid/RPE/Retina whole mount staining was recently developed and reported. The method can be further combined and optimized with melanin bleaching and tissue clearing. Here, we describe steps of both non-pigmented and pigmented mouse Sclerochoroid/RPE/Retina whole mount including eyeball preparation, staining, mounting and confocal scanning. In

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[摘要]  [摘要]视网膜色素上皮(RPE)和感光细胞之间的视网膜下层是与年龄相关的黄斑变性等疾病进程中涉及炎症和血管生成的区域。由于需要在视网膜和RPE之间分离,因此整个安装(视网膜和RPE)的当前方案无法解决视网膜下层的完整视图,然后对每个单独的组织进行染色。最近开发并报道了非分离的巩膜脉络膜/ RPE /视网膜整装染色。该方法可以与黑色素漂白和组织清除进一步结合和优化。在这里,我们描述了非色素和色素小鼠硬化脉络膜的步骤/ RPE /视网膜整个安装,包括眼球准备,染色,安装和共聚焦扫描。此外,我们提出了巩膜脉络膜/ RPE /视网膜整个支架中的RPE,视网膜下小胶质细胞和邻近的感光体的共聚焦图像。

[背景]在眼睛视网膜是由视网膜色素上皮(RPE)包围,脉络膜和巩膜。通常,在整装染色中,视网膜组织与脉络膜/ RPE分开,并且分开的视网膜和脉络膜/ RPE的每个部分都被染色。因此,视网膜或脉络膜/ RPE的单独的整体染色不能解决完整的视网膜下信息。最近,开发了脉络膜/ RP E /视网膜整个安装方案(Kim等,2016; ...

Triacylglycerol Measurement in HeLa Cells
Author:
Date:
2020-12-20
[Abstract]  

Lipid droplets store triacylglycerols (triglycerides) and sterol esters to regulate lipid and energy homeostasis. Triacylglycerol measurement is often performed during the investigation of lipid droplet formation and growth. This protocol describes a reliable method using a fluorometric lipid quantification kit to measure triacylglycerols extracted from HeLa cells, which were treated with oleic acid to trigger the formation of lipid droplets. The lipid quantification kit employs a lipid-binding molecule that emits bright fluorescence only when bound to extracted triacylglycerols, whose content can be quantified by a simple fluorescence readout.

[摘要]  [摘要]脂滴商店吨riacylglycerols (甘油三酯)和甾醇酯调节脂质和能量稳态。通常在研究脂质滴的形成和生长过程中进行三酰基甘油的测量。该协议描述了一种使用荧光脂质定量试剂盒测量从HeLa细胞提取的三酰基甘油的可靠方法,该甘油经油酸处理后触发了脂质滴的形成。脂质定量试剂盒采用的脂质结合分子仅在与提取的三酰基甘油结合时才发出明亮的荧光,其含量可以通过简单的荧光读数来定量。

[背景]脂滴(LD)的是具有特征功能独特的细胞器。与常见的细胞器不同,每个LD由磷脂的单层定义,该磷脂包裹着中性脂质(如三酰甘油(TAG),也称为甘油三酸酯)和甾醇酯的核心。作为存储细胞器,LDs在脂质和能量的动态平衡以及其他细胞事件中起主要作用。通过以TAG的形式存储过量的脂肪酸,LDs可以防止脂毒性并保护细胞免受氧化应激。重要的是,LDs几乎与所有其他细胞器接触并调节关键的细胞过程,包括膜运输,蛋白质更新和病毒感染(Olzmann和Carvalho,2018; Gao等,2019)。此外,LD表面装饰有各种蛋白质,这些蛋白质参与TAG的合成和水解以及LD与其他细胞器之间的脂质运输(Kory等,2016; Kumar等,2018; Du等。 。,2020)。

真核细胞具有完善的途径来合成LDs中存储的TAG ...

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