Spectral flow cytometer

光谱流式细胞仪

• 批准号: 499549016
• 金额: --
• 依托单位: Ludwig-Maximilians-Universität München (LMU)
• 依托单位国家: 德国
• 项目类别: Major Research Instrumentation
• 财政年份: 2022
• 资助国家: 德国
• 起止时间: 2021-12-31 至 无数据
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/499549016?language=en
• 关键词: Spectral; flow; cytometer

This request is for the addition of a spectral flow cytometer to the Core Facility for Flow Cytometry at the Campus Innenstadt (LMU Klinikum) to meet the needs for cutting-edge research in the coming years. The Core Facility is used by research groups focusing on immunology, tumor immunology, autoimmunity and vaccinology. With conventional flow cytometers, technically only a limited number of colors can be analyzed simultaneously. In the Core Facility three flow cytometers are currently in use. The most modern instrument in the facility allows the discrimination of 16 parameters. This is not sufficient for the measurement of many newly defined cell types or an exploratory analysis of immune cells. Spectral analysis clearly shifts this limitation. With currently 40 different colors that can be measured simultaneously, a similar order of magnitude is achieved as with the much more expensive and complex mass cytometry. By measuring many parameters simultaneously, even patient samples with limited cell numbers can be defined with high resolution. Spectral analysis offers the following advantages: - the significantly improved sensitivity allows improved resolution of low-expressed surface antigens by photon measurement in the whole spectrum;- similar fluorochromes can also be distinguished by whole spectrum analysis; - fluorescent proteins can be better differentiated, significantly increasing experimental possibilities;- cell population-dependent subtraction of autofluorescence offers a distinct advantage in the analysis of low-expressed markers on cells with strong autofluorescence, e.g. granulocytes;- in addition to the advantages described above, the requested device has an integrated plate reader so that a large number of samples (e.g., in a 96-well plate) can be analyzed automatically. This increases reproducibility and accelerates the throughput rate.

这一请求是为了在校园Innenstadt（LMU Klinikum）的流式细胞术核心设施中增加一台光谱流式细胞仪，以满足未来几年尖端研究的需求。核心设施由专注于免疫学，肿瘤免疫学，自身免疫学和疫苗学的研究小组使用。使用传统的流式细胞仪，技术上只能同时分析有限数量的颜色。核心设施目前使用三台流式细胞仪。该设施中最现代化的仪器可以识别16个参数。这不足以测量许多新定义的细胞类型或免疫细胞的探索性分析。光谱分析显然改变了这一限制。由于目前可以同时测量40种不同的颜色，因此可以实现与昂贵得多且复杂得多的质谱仪类似的数量级。通过同时测量多个参数，即使是细胞数量有限的患者样本也可以高分辨率进行定义。光谱分析提供了以下优点：-显著提高的灵敏度允许通过全光谱中的光子测量来提高低表达表面抗原的分辨率;-类似的荧光染料也可以通过全光谱分析来区分; -荧光蛋白可以更好地区分，显著增加实验可能性;- 自发荧光的细胞群体依赖性扣除在分析具有强自发荧光的细胞（例如粒细胞）上的低表达标志物方面提供了明显的优势;- 除了上述优点之外，所请求的设备具有集成的板读取器，使得大量样品（例如，在96孔板中）可以自动分析。这提高了再现性并加快了生产率。