Spectral Flow Cytometer

光谱流式细胞仪

• 批准号: 543853730
• 金额: --
• 依托单位: Medizinische Hochschule Hannover (MHH)
• 依托单位国家: 德国
• 项目类别: Major Research Instrumentation
• 财政年份: 2024
• 资助国家: 德国
• 起止时间: 2023-12-31 至 无数据
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/543853730?language=en
• 关键词: Spectral; Flow; Cytometer

An essential part of the research in the Department of General, Visceral and Transplant Surgery is the characterization and purification of cell populations from blood samples, dissociated tissue material, primary cell cultures or immortalized cell lines on the basis of their surface proteins in the murine, porcine or human system. For the research of the working group, a flow cytometer with a maximum of 16 parameters to be measured was available in Berlin. The use of the flow cytometer by the applicant's working group was constantly at a high level (approx. 650 - 1000 hours/year). The FACS device available at the MHH, in the research laboratory of the Clinic for General, Visceral and Transplant Surgery, can only measure 6 different parameters (4 colors + SSC, FSC). A new multispectral flow cytometer is indispensable for the continuation of the research activities. The requested multispectral flow cytometer is an instrument that measures the entire emitted fluorescence spectrum for each fluorochrome used, rather than individual spectral regions. This allows the use of fluorochromes with closely adjacent emission spectra. A multispectral flow cytometer equipped with five lasers can measure more than 40 parameters in parallel, allowing the analysis of complex cell phenotypes. In addition, signal quality and sensitivity can be improved by subtracting autofluorescence based on cell population. This is especially important for tissue-specific autofluorescence, but also for granulocytes or fluorescent proteins. A multispectral characterization of the cells under investigation allows a significant reduction in the number of samples required and thus in the amount of material and time needed. In view of the often very limited samples available, e.g. from murine animal models or patient biopsies, such an instrument allows a maximum gain of information. Populations within a cell mixture can be characterized in detail. Patient samples with a limited number of cells can be defined with high resolution and animal experiments can be reduced according to the 3R principles. We are requesting a multispectral flow cytometer that will meet the needs of cutting-edge research in the coming years. The requested instrument will allow us to continue the research of the past years, to expand existing research approaches and to enable new research projects.

普通、内脏和移植外科研究的一个重要部分是从血液样本、分离的组织材料、原代细胞培养或永生化细胞系中根据它们在小鼠、猪或人类系统中的表面蛋白来表征和纯化细胞群。对于工作组的研究，柏林提供了一台最多可测量16个参数的流式细胞仪。申请人的工作组对流式细胞仪的使用一直处于较高水平(约650-1000小时/年)。在全科、内脏和移植外科诊所的研究实验室，MHH提供的FACS设备只能测量6种不同的参数(4种颜色+SSC、FSC)。为了继续开展研究活动，新的多光谱流式细胞仪是必不可少的。所要求的多光谱流式细胞仪是一种测量每个使用的荧光色素的整个发射荧光光谱的仪器，而不是单独的光谱区域。这允许使用发射光谱相近的荧光染料。配备五个激光的多光谱流式细胞仪可以并行测量40多个参数，从而能够分析复杂的细胞表型。此外，信号质量和灵敏度可以通过减去基于细胞数量的自体荧光来改善。这对组织特异性的自体荧光尤其重要，对粒细胞或荧光蛋白也是如此。对正在调查的细胞进行多光谱表征可以大大减少所需样品的数量，从而减少所需材料的数量和所需的时间。鉴于可获得的样本往往非常有限，例如来自小鼠动物模型或患者活检组织的样本，这种仪器允许最大限度地获得信息。细胞混合物中的种群可以被详细描述。细胞数量有限的患者样本可以用高分辨率定义，动物实验可以根据3R原则减少。我们需要一种多光谱流式细胞仪，以满足未来几年尖端研究的需要。所要求的仪器将使我们能够继续过去几年的研究，扩大现有的研究方法，并使新的研究项目成为可能。