| Histological and Immunohistochemical Examination of Stem Cell Proliferation and Reepithelialization in the Wounded Skin
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Author:
Date:
2021-01-20
[Abstract] The skin is the largest organ that protects our body from the external environment and it is constantly exposed to pathogenic insults and injury. Repair of damage to this organ is carried out by a complex process involving three overlapping phases of inflammation, proliferation and remodeling. Histological analysis of wounded skin is a convenient approach to examine broad alterations in tissue architecture and investigate cells in their indigenous microenvironment. In this article we present a protocol for immunohistochemical examination of wounded skin to study mechanisms involved in regulating stem cell activity, which is a vital component in the repair of the damaged tissue. Performing such histological analysis enables the understanding of the spatial relationship between cells that ...
[摘要] [摘要]皮肤是保护我们的身体免受外界环境影响最大的器官,并且不断受到病原体的伤害和伤害。修复该器官的水坝年龄是通过复杂的过程进行的,该过程涉及炎症,增殖和重塑的三个重叠阶段。对受伤的皮肤进行组织学分析是检查组织结构的广泛变化并研究其固有微环境中的细胞的便捷方法。在本文中,我们提出了一种用于受伤皮肤的免疫组织化学检查的方案,以研究参与调节干细胞活性的机制,这是修复受损组织的重要组成部分。进行这种组织学分析可以了解在特殊伤口微环境中相互作用的细胞之间的空间关系。本文所述的分析工具,允许邻近伤口和重新程度的干细胞的再生能力的定量测量-伤口闭合过程中的上皮。这些协议可以适用于研究受伤皮肤内的众多细胞过程和细胞类型。
[背景]皮肤具有巨大的再生能力,这归因于位于整个器官不同部位的众多常驻干细胞种群的存在。免疫组织学方法与目标细胞的遗传标记相结合,已大大提高了体内对干细胞调节和组织稳态的认识(Blanpain等,2004)。这些方法已用于鉴定与毛囊相关的多个干细胞群体,包括位于毛发隆突中的表皮和黑素细胞干细胞,次级毛发芽中的快速循环祖细胞以及真皮鞘中存在的真皮干细胞(Greco等。,2009;拉赫马尼等人,2020)。这些干细胞具有自我更新,多能性和高度增殖性,可在生理性毛发循环过程中再生组织,并在伤口愈合过程中修复损伤。引人注目的是,由于这些细胞分裂并迅速向伤口部位进展,使上皮中的干细胞对皮肤损伤迅速做出反应,从而使伤口重新上皮化(Ito等人,2005)。 ...
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| 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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