| Optimized Immunostaining of Embryonic and Early Postnatal Mouse Brain Sections
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Author:
Date:
2021-01-05
[Abstract] The mammalian neocortex, the outer layer of the cerebrum and most recently evolved brain region, is characterized by its unique areal and laminar organization. Distinct cortical layers and areas can be identified by the protein expression of graded transcription factors and molecular determinants that define the identity of different projection neurons. Thus, specific detection and visualization of protein expression is crucial for assessing the identity of neocortical neurons and, more broadly, for understanding early and late developmental mechanisms and function of this complex system. Several immunostaining/immunofluorescence methods exist to detect protein expression. Published protocols vary with regard to subtle details, which may impact the final outcome of the immunofluorescence. ...
[摘要] [摘要]哺乳动物的新皮层是大脑的外层，也是最近进化的大脑区域，其特征在于其独特的区域和层状组织。不同的皮质层和区域可以通过分级转录因子的蛋白质表达和定义不同投射神经元身份的分子决定簇来鉴定。因此，蛋白质表达的特异性检测和可视化对于评估新皮层神经元的身份至关重要，更广泛地来说，对于理解这个复杂系统的早期和晚期发育机制和功能至关重要。一些 存在免疫染色/免疫荧光方法来检测蛋白质表达。已发布的方案在细节方面有所不同，可能会影响免疫荧光的最终结果。在这里，我们提供了详细的协议，适用于低温恒温器薄切片和厚振动切片器薄切片，该协议已成功地开发了针对新皮质发育关键分子的抗体。从早期的技术措施测距的大脑采集到的图像分析和统计，我们包括有关样品包容和每一个细节部分ING，幻灯片存储和旨在减少非特异性背景最佳抗体稀释。在实验室中常规使用的，我们的背景优化的免疫染色协议允许区域的有效检测-和层-独特新皮层投射神经元的特定的分子决定因素。
图形摘要：
优化的小鼠脑切片免疫染色方案的工作流程图。一。一个流程图的不同步骤的优化免疫染色协议薄低温恒温器和厚振动切片。乙。例如，用于在薄冠状免疫染色对SATB2和CTIP2部（20 μ ...
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| Isolation of Splenic Dendritic Cells Using Fluorescence-activated Cell Sorting
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Author:
Date:
2015-03-05
[Abstract] The spleen is a vastly vasculated organ and consists of a complex organized network of innate and adaptive immune cells. This permits the specialized functions of the spleen such as antibacterial and antifungal immunity and iron metabolism among others (Mebius and Kraal, 2005). Different dendritic cell (DC) subsets reside in the spleen and can be defined by the expression of unique surface markers. These DC subsets are recognized to perform non-redundant functions in the immune system (Merad et al., 2013). In our recent study, we found that Inositol Requiring Enzyme (IRE)-1 is specifically activated in splenic CD8a+ DCs. Furthermore, loss of X-box binding protein (XBP)-1 – the transcription factor regulated by IRE-1 – resulted in defective cross-presentation of dead ...
[摘要] 脾脏是一个巨大的血管的器官，由一个复杂的有组织的网络的先天和适应性免疫细胞组成。 这允许脾的特殊功能，例如抗菌和抗真菌免疫和铁代谢等（Mebius和Kraal，2005）。 不同的树突状细胞（DC）子集驻留在脾脏中，并且可以通过表达独特的表面标志物来定义。 认识到这些DC亚类在免疫系统中执行非冗余功能（Merad等人，2013）。 在我们最近的研究中，我们发现肌醇需求酶（IRE）-1在脾CD8a + DC中特异性激活。 此外，X-box结合蛋白（XBP）-1（由IRE-1调节的转录因子）的缺失导致脾CD8a + DC（Osorio em> et al。，2014）。 该协议允许分离用于实验使用的特异性DC亚群ex-vivo 。
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| Allergen Sensitization and Challenge to Ovalbumin
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Author:
Date:
2013-04-05
[Abstract] This protocol describes the sensitization and challenge of mice with ovalbumin for use as an acute murine model of asthma. This protocol induces reproducible airway inflammation and remodelling, and bronchial hyperresponsiveness to methacholine as measured by barometric plethysmography, as well as by the Flexivent® technique in Balb/c mice.
[摘要] 该协议描述了用卵清蛋白作为哮喘的急性鼠模型的小鼠的致敏和攻击。 该协议诱导可重复的气道炎症和重塑，以及通过气压体积描记术以及通过Flexb/c小鼠中的Flexivent 技术测量的对乙酰甲胆碱的支气管高反应性。
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