| Combination of Fluorescent in situ Hybridization (FISH) and Immunofluorescence Imaging for Detection of Cytokine Expression in Microglia/Macrophage Cells
|
|
Author:
Date:
2017-11-20
[Abstract] Microglia and macrophage cells are the primary producers of cytokines in response to neuroinflammatory processes. But these cytokines are also produced by other glial cells, endothelial cells, and neurons. It is essential to identify the cells that produce these cytokines to target their different levels of activation. We used dual RNAscope® fluorescence in situ hybridization (FISH) and immunohistochemistry (IHC) techniques to visualize the mRNA expression pattern of pro- and anti-inflammatory cytokines in microglia/macrophages cells. Using these methods, we can associate one mRNA to specific cell types when combining with different cellular markers by immunofluorescence. Results from RNAscope® probes IL-1β, TNFα, TGFβ, IL-10 or Arg1, showed colocalization ...
[摘要] 小神经胶质细胞和巨噬细胞是响应神经炎症过程的细胞因子的主要生产者。但是这些细胞因子也是由其他神经胶质细胞，内皮细胞和神经元产生的。鉴定产生这些细胞因子的细胞以靶向其不同水平的活化是至关重要的。我们使用双RNAscope荧光原位杂交（FISH）和免疫组织化学（IHC）技术来观察小胶质细胞/巨噬细胞中促炎细胞因子和抗炎细胞因子的mRNA表达模式细胞。使用这些方法，我们可以联合一个mRNA与特定的细胞类型时，通过免疫荧光与不同的细胞标志物结合。来自RNAscope探针的结果IL-1β，TNFα，TGFβ，IL-10或Arg1显示与小胶质细胞/巨噬细胞抗体的共定位。这些靶标探针显示出足够的灵敏度和特异性来检测mRNA表达。新的FISH检测技术结合免疫组化技术将有助于共同确定蛋白质和mRNA的定位，以及提供可靠的mRNA表达水平的量化。
【背景】mRNA原位杂交技术是一种有用的工具，其允许以细胞依赖性方式特异性和选择性标记脑切片中的RNA序列（Grabinski等人，2015 ...
|
|
|
| Fluoro-Jade B Staining for Neuronal Cell Death
|
|
Author:
Date:
2016-01-05
[Abstract] Fluoro-Jade is a fluorescent derivative used for histological staining of degenerating neurons. This technique is simple and sensitive enough to label distal dendrites, axons, axon terminals as well as neuronal bodies. Fluoro-Jade has excitation and emission peak of 480 and 525 nanometer respectively. It can be visualized using a fluorescein/FITC filter. Some reports have demonstrated that Fluoro-Jade can also be useful to detect glial cell death (Anderson et al., 2013; Damjanac et al., 2007).
[摘要] Fluoro-Jade是用于退化神经元的组织学染色的荧光衍生物。 这种技术是简单和足够敏感到标记远端树突，轴突，轴突终端以及神经元体。 翡翠的激发和发射峰分别为480和525纳米。 它可以使用荧光素/FITC过滤器可视化。 一些报道已经证明氟 - 玉也可以用于检测胶质细胞死亡（Anderson等人，2013; Damjanac等人，2007）。
|
|
|
| Mouse Models of Uncomplicated and Fatal Malaria
|
|
Author:
Date:
2015-07-05
[Abstract] Mouse models have demonstrated utility in delineating the mechanisms underlying many aspects of malaria immunology and physiology. The most common mouse models of malaria employ the rodent-specific parasite species Plasmodium berghei, P. yoelii, and P. chabaudi, which elicit distinct pathologies and immune responses and are used to model different manifestations of human disease. In vitro culture methods are not well developed for rodent Plasmodium parasites, which thus require in vivo maintenance. Moreover, physiologically relevant immunological processes are best studied in vivo. Here, we detail the processes of infecting mice with Plasmodium, maintaining the parasite in vivo, and monitoring parasite levels ...
[摘要] 小鼠模型已经证明了用于描绘疟疾免疫学和生理学的许多方面的机制的效用。 最常见的疟疾小鼠模型使用啮齿动物特异性寄生虫物种伯氏疟原虫 。 yoelii 和 P。 chabaudi ，其引起不同的病理学和免疫应答，并用于模拟人类疾病的不同表现。 体外培养方法对于啮齿动物疟原虫寄生虫不是很好发展，因此需要在体内维持。 此外，生理相关的免疫过程最好在体内研究。 在这里，我们详细的感染小鼠与疟原虫，维持寄生虫体内，并监测寄生虫水平和健康参数整个感染的过程。
|
|
|