Flow cytometer (5 laser system)

免疫学和神经免疫学研究的一个关键方法是通过流式细胞术分析不同的细胞群。在免疫学研究中，大多数细胞群体的多参数分析都是使用高端流式细胞仪进行的，这使得对这些群体的分析变得容易和详细。流式细胞术使研究人员不仅可以通过荧光团偶联抗体标记不同的标记，还可以同时分析不同的染料和荧光蛋白。尤其是在神经免疫学新的拓展领域中，对不同的中枢神经系统驻留和相关细胞群体进行详细的多参数分析变得越来越重要。最近的研究指出，在健康和患病的中枢神经系统中，这些免疫细胞群具有一定的异质性。这些新的发现突出了这些中枢神经系统免疫细胞群的复杂性、动力学和可塑性，进一步证明了在流式细胞术中通过应用多参数面板在不同的环境下进行分析来尽可能好地解剖这些细胞群的迫切需要。具有五个激光的最先进的流式细胞仪将为我们的研究在选择荧光团、染料和荧光蛋白方面提供一个新的维度和灵活性。五条激光线路提供了广泛的颜色选择，并允许将用于分析的标记分布在不同的激光线路上；因此可以减少不同通道之间的溢出效应。在五路激光流式细胞仪中，许多常见的标记和荧光蛋白可以被两条不同的激光谱线激发，而额外的一条紫外激光将为标记分析提供新的通道。这将导致对单独通道中的信号进行更灵敏的检测，并在通道之间实现更高的分辨率。通过使用强度可调的激光线路，可以进一步减小溢出效应或微弱信号的损失。到目前为止，研究所有一台三激光流式细胞仪(10个参数)，但这种流式细胞仪不足以满足计划中的研究项目和复杂的研究问题。迫切需要一个新的五激光系统，使我们能够进一步推进这些项目，并在我们的研究项目中取得新的进展。因此，我们想申请一个新的五激光流式细胞仪与这一建议。

A key method of immunological, as well as neuroimmunological, research is the analysis of different cell populations via flow cytometry. In immunological research, most multi-parameter analyses of cell populations are performed with high end flow cytometers which allow an easy and detailed analysis of these populations. Flow cytometry allows the researcher to not only label distinct markers via fluorophore-coupled antibodies, but also to analyze different dyes and fluorescent proteins at once. Especially in the new expanding field of neuroimmunology, detailed multi-parameter analysis of different CNS-resident and –associated cell populations become more and more important. Recent studies point to a certain heterogeneity within these immune cell populations in the healthy and diseased CNS. These new findings highlight the complexity, dynamics and plasticity of these CNS immune cell populations, further demonstrating the urgent need of dissecting these cell populations as good as possible in flow cytometry by applying multi-parameter panels for analysis in various settings.A state of the art flow cytometer with five lasers would offer our research a new dimension and flexibility in the choice of fluorophores, dyes and fluorescent proteins. Five laser lines offer a wide spectrum of color choices and allow distributing the markers for the analysis on different laser lines; therefore spill-over effects between different channels can be reduced. Many common markers and fluorescent proteins can be excited by two different laser lines in a five laser flow cytometer and an additional UV-laser would offer new channels for marker analysis. This will lead to a more sensitive detection of the signals in separate channels and a higher resolution between the channels. Spill-over effects or the loss of weak signals can be further diminished by using laser lines with adjustable intensities. A three laser flow cytometer (10 parameters) is so far available in the institute; however this flow cytometer is not sufficient for the planned research projects and the complexity of the research questions. There is an urgent need for a new five laser system to allow us to further pursue the projects and to achieve new advances in our research projects. Therefore, we would like to apply for a new five laser flow cytometer with this proposal.