Spectral Flow Cytometer

一般内脏和移植外科部门研究的一个重要部分是在小鼠，猪或人类系统中基于其表面蛋白质的血液样本，解离组织材料，原代细胞培养或永生化细胞系中细胞群体的特征和纯化。为了工作组的研究，柏林有一台最多可测量16个参数的流式细胞仪。申请人的工作小组对流式细胞仪的使用一直处于较高水平(约。650 - 1000小时/年)。MHH在普通、内脏和移植手术诊所的研究实验室中提供的FACS设备只能测量6种不同的参数（4种颜色+ SSC， FSC）。一种新型的多光谱流式细胞仪是研究工作继续进行的必要条件。所要求的多光谱流式细胞仪是一种测量所使用的每种荧光染料的整个发射荧光光谱的仪器，而不是单个光谱区域。这就允许使用发射光谱非常接近的荧光染料。配备5个激光器的多光谱流式细胞仪可以并行测量40多个参数，允许分析复杂的细胞表型。此外，信号质量和灵敏度可以通过基于细胞群减去自身荧光来提高。这对组织特异性自身荧光尤其重要，但对粒细胞或荧光蛋白也很重要。对所研究的细胞进行多光谱表征可以显著减少所需样品的数量，从而减少所需的材料和时间。鉴于通常可获得的样本非常有限，例如来自小鼠动物模型或患者活检，这种仪器可以最大限度地获得信息。细胞混合物内的种群可以被详细地描述。细胞数量有限的患者样本可以用高分辨率定义，根据3R原则可以减少动物实验。我们正在寻求一种多光谱流式细胞仪，以满足未来几年前沿研究的需要。所要求的工具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。

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.