5-laser spectral -analysis flow cytometer

在过去的2-3年中，波恩流式细胞术的应用已经明显转向多参数分析。3激光系统，其中一些已经使用了10多年，并在一个持续的高水平很长一段时间（平均约10年）。1200小时/年），而两台5激光设备（LSR Fortessa）的使用稳步增加。在2018年夏天购买了第二台5激光分析仪后，几乎从一开始就实现了与现有设备类似的高利用率。这两款设备现在都接近其利用率限制，在过去的一年里几乎有2000小时的预订时间。因此，科学家们经常被迫进行测量，直到深夜。光谱流式细胞仪旨在减轻现有基础设施的负担。此外，光谱操作流式细胞仪具有相同的设备和激发光源，提供了传统流式细胞仪的优势。显著扩展了多参数流式细胞仪的应用范围。这包括使用具有密切相关的发射光谱的染料或通过“光谱解混”和“自体荧光提取”对整个光谱进行后续处理，以提高信号质量，从而提高灵敏度。荧光蛋白在模式生物中的应用，与之相关的组织特异性自体荧光和由此在随后的多参数免疫表型分析中产生的问题是一个很好的解决方案，新的应用，例如在无标记表型领域和生物信息学中大量数据集的处理是当前研究的主题。对核心设施用户的调查能够确定对于大多数潜在的未来用户来说，由于上述原因，来自该中心的光谱流式细胞术是核心设施基础设施的首选技术扩展。受访的科学家涵盖了大学医学和科学研究的最多样化的领域。这里要提到的主要是：免疫学，神经科学，癌症研究，传染病和心脏病学。本申请中总结的许多研究项目将直接受益于所提到的光谱分析的优点，或者实际上只能使用这种设备进行。这方面的详细信息见第3点“研究用途”。

In the last 2-3 years there has been a noticeable shift in applications in flow cytometry towards multiparametric analyzes in Bonn. The 3-laser systems, some of which have been in use for over 10 years and for a long time at a consistently high level (average approx. 1200 hours / year), whereas the use of the two 5-laser devices (LSR Fortessa) has steadily increased. After purchasing the second 5-laser analyzer in the summer of 2018, a similarly high level of utilization as the existing device was achieved almost right from the start. Both devices are now close to their utilization limitalmost 2000 hours of booked time in the last year. The scientists are therefore often forced to carry out their measurements until late at night. The spectral flow cytometer applied for here is intended to relieve the existing infrastructure.In addition, spectrally operating flow cytometers, with identical equipment with excitation light sources, offer advantages over conventional flow cytometersSignificantly expand the range of applications for multiparametric flow cytometry. This includes the use of dyes with closely related emission spectra or the subsequent processing of the entire spectrum through “spectral unmixing” and “autofluorescence extraction” to improve the signal quality and thus the sensitivity. The use of fluorescent proteins in model organisms, the tissue-specific autofluorescence associated therewith and the problems resulting therefrom in a subsequent multi-parametric immunophenotyping are a well-known example of this.Novel applications e.g. in the field of markerless phenotyping and theProcessing of the extensive data sets in bioinformatics are the subject of thiscurrent research.A survey of the users of the Core Facility was able to determine that for a large majority of potential future users, the spectral flow cytometry from therepresents the preferred technological expansion of the infrastructure of the core facility for the reasons mentioned above. The interviewed scientists cover the most diverse areas of medical and scientific research at the university. The main ones to be mentioned here are: immunology, neurosciences, cancer research, infectious diseases and cardiology. Many of the research projects summarized in this application would benefit directly from the advantages of spectral analysis mentioned or can in fact only be carried out with such a device. Detailed information on this can be found under point 3 "Use in research".