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4’,6-diamidino-2-phenylindole (DAPI)

DAPI

Company: Sigma-Aldrich
Catalog#: D9542
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Direct Reprogramming of Mouse Embryonic Fibroblasts to Conventional Type 1 Dendritic Cells by Enforced Expression of Transcription Factors
Author:
Date:
2020-05-20
[Abstract]  Ectopic expression of transcription factor combinations has been recently demonstrated to reprogram differentiated somatic cells towards the dendritic cell (DC) lineage without reversion to a multipotent state. DCs have the ability to induce potent and long-lasting adaptive immune responses. In particular, conventional type 1 DCs (cDC1s) excel on antigen cross-presentation, a critical step for inducing CD8+ T cell cytotoxic responses. The rarity of naturally occurring cDC1s and lack of in vitro methodologies for the generation of pure cDC1 populations strongly hinders the study of cDC1 lineage specification and function. Here, we describe a protocol for the generation of induced DCs (iDCs) by lentiviral-mediated expression of the transcription factors PU.1, IRF8 and ... [摘要]  [摘要] 转录因子组合的异位表达最近被证明可以将分化的体细胞重编程为树突状细胞(DC)谱系,而不会回复到多能状态。DC具有诱导有效和持久的适应性免疫应答的能力。在特定的常规类型1的DC(cDC1s)练成抗原交叉呈递,用于诱导CD8的关键步骤+ T细胞的细胞毒性应答。天然存在的cDC1的稀有性和缺乏用于生成纯cDC1群体的体外方法论,严重阻碍了对cDC1谱系规格和功能的研究。在这里,我们描述了用于生成感应DC(iDC)的协议 慢病毒介导的转录因子PU.1,IRF8和BATF3在小鼠胚胎成纤维细胞中的表达。iDC 在9天内获得DC形态,cDC1表型和转录特征。使用此协议生成的iDC 具有对炎症刺激,吞噬死细胞,将抗原加工并交叉呈递给CD8 + T细胞的功能。DC重新编程提供了一个简单易处理的系统,可以生成大量的cDC1类细胞用于高内涵筛选,从而开辟了新途径,可以更好地了解cDC1的规格和功能。将来,在成纤维细胞中忠实诱导cDC1命运可能会导致产生患者特定的疫苗接种DC。

[背景技术树突状细胞(DC)是专业的抗原呈递细胞,专门用于识别,加工和呈递T细胞抗原,在诱导适应性免疫应答和免疫记忆中起关键作用(Me rad 等,2013)。DC可以分为4个主要子集:浆细胞样DC(pDC ),大量1型干扰素的产生者,循环单核细胞衍生的单核细胞衍生DC 和常规DC(cDC ...

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。

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