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DAPI

Company: Sigma-Aldrich
Catalog#: D9542
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Qualitative in vivo Bioluminescence Imaging
Author:
Date:
2018-09-20
[Abstract]  Bioluminescence imaging (BLI) technology is an advanced method of carrying out molecular imaging on live laboratory animals in vivo. This powerful technique is widely-used in studying a variety of biological processes, and it has been an ideal tool in exploring tumor growth and metastatic spread in real-time. This technique ensures the optimal use of laboratory animal resources, particularly the ethical principle of reduction in animal use, given its non-invasive nature, ensuring that ongoing biological processes can be studied over time in the same animal, without the need to euthanize groups of mice at specific time points. In this protocol, the luciferase imaging technique was developed to study the effect of co-inoculating pericytes (contractile, αSMA+ mesenchymal ... [摘要]  生物发光成像(BLI)技术是一种在体内实验室动物上进行分子成像的先进方法。 这种强大的技术广泛应用于研究各种生物过程,是实时探索肿瘤生长和转移扩散的理想工具。 该技术确保实验室动物资源的最佳利用,特别是减少动物使用的伦理原则,考虑到其非侵入性,确保可以在同一动物中随时间研究正在进行的生物过程,而无需安乐死 小鼠在特定的时间点。 在该方案中,开发了荧光素酶成像技术以研究共同接种周细胞(收缩性,αSMA + 间充质干细胞样细胞,位于微血管内的细胞)对生长和转移性扩散的影响。 卵巢癌使用侵袭性卵巢癌细胞系-OVCAR-5-作为例子。

【背景】生物发光成像(BLI)的原理是基于相对简单的生化过程的发光特性,即,荧光素酶介导的分子底物荧光素氧化产生光。在癌症研究中,BLI是一种流行的工具(Contag et ...

Embryonic Intravitreous Injection in Mouse
Author:
Date:
2018-07-20
[Abstract]  Axons of retinal ganglion cells (RGCs) relay visual information from the retina to lateral geniculate nucleus (LGN) and superior colliculus (SC), which are two major image-forming visual nuclei. Wiring of these retinal projections completes before vision begins. However, there are few studies on retinal axons at embryonic stage due to technical difficulty. We developed a method of embryonic intravitreous injection of dyes in mice to visualize retinal projections to LGN and SC. This study opens up the possibility of understanding early visual circuit wiring in mice embryos. [摘要]  视网膜神经节细胞(RGC)的轴突将视觉信息从视网膜传递到外侧膝状核(LGN)和上丘(SC),这是两个主要的成像视觉核。 在视力开始之前完成这些视网膜投射的布线。 然而,由于技术困难,很少有关于胚胎期视网膜轴突的研究。 我们开发了一种胚胎玻璃体内注射染料的方法,用于观察LGN和SC的视网膜投射。 这项研究开辟了理解小鼠胚胎早期视觉电路布线的可能性。

【背景】早在胚胎第14.5天(E14.5)小鼠中,视网膜轴突开始向LGN和SC投射。 为了研究胚胎中的轴突投射,需要在注射手术前保持胚胎存活至少10小时,玻璃内注射染料。 以前的研究表明,胚胎可以使用含氧小鼠血清进行体外培养。 然而,营养多样性,氧饱和度和温度调节等问题会影响胚胎的生理状况。 在我们的研究中,我们通过胚胎中的子宫壁和眼睑进行玻璃体内注射,并在注射后将它们保存在子宫中。 从E15.5到E18.5很好地标记了对LGN和SC的视网膜轴突投射。

Quantification of Extracellular Double-stranded RNA Uptake and Subcellular Localization Using Flow Cytometry and Confocal Microscopy
Author:
Date:
2018-06-20
[Abstract]  Double-stranded RNA is a potent pathogen-associated molecular pattern (PAMP) produced as a by-product of viral replication and a well-known hallmark of viral infection. Viral dsRNAs can be released from infected cells into the extracellular space and internalized by neighboring cells via endocytosis. Mammals possess multiple pattern recognition receptors (PRRs) capable of detecting viral dsRNAs such as endosomal toll-like receptor 3 (TLR3) and cytosolic RIG-I-like receptors (RLRs) which lead to the production of type I interferons (IFNs). Thus, intracellular localization of viral dsRNA can provide insight into the downstream signaling pathways leading to innate immune activation. Here, we describe a quantitative method for measuring extracellular dsRNA uptake and visualizing subcellular ... [摘要]  双链RNA是一种有效的病原体相关分子模式(PAMP),作为病毒复制的副产物和病毒感染的众所周知的标志产生。 病毒dsRNA可以从感染的细胞释放到细胞外空间并通过胞吞作用被邻近细胞内化。 哺乳动物具有能够检测导致产生I型干扰素(IFN)的病毒dsRNA(例如内体Toll样受体3(TLR3)和胞质RIG-1样受体(RLR))的多模式识别受体(PRR)。 因此,病毒dsRNA的细胞内定位可以提供对导致先天免疫激活的下游信号传导途径的了解。 在这里,我们描述了一种测量细胞外dsRNA摄取和分别通过流式细胞仪和共聚焦显微镜观察内化dsRNA的亚细胞定位的定量方法。

【背景】双链RNA(dsRNA)是病毒复制的常见副产物,通过产生I型干扰素(IFN)和其他促炎细胞因子(Nellimarla和Mossman,2014)是抗病毒免疫的有效激活剂。病毒的dsRNA通过TLR3核内体内所感测(松本等人,2003年)或在由RIG-I样受体(RLRS),RIG-I和MDA-5(加藤等人,2006)。在裂解感染,这些dsRNA可以被释放到细胞外空间,它们结合于相邻小区,如A类清道夫受体(SR-A)和RAFTLIN表面受体,并且随后经由网格蛋白介导的内吞作用内在化(伊藤制 2008; DeWitte-Orr等人,2010; Watanabe等人,2011; Dansako等人,, ...

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