German Rheumatology Research Center (DRFZ) - Flow Cytometry Core Facility, Charitéplatz 1 (Virchowweg 12), 10117, Berlin, Germany.
APE Angewandte Physik und Elektronik GmbH, Plauener Straße 163-165 / Haus N, 13053, Berlin, Germany.
Commun Biol. 2024 Aug 30;7(1):1063. doi: 10.1038/s42003-024-06759-5.
Flow cytometry is a key technology for the analysis and sorting of cells or particles at high throughput. Conventional and current flow cytometry is primarily based on fluorescent stains to detect the cells of interest. However, such stains also have disadvantages, as their effect on cells must be carefully tested to avoid effects on the results of the experiments. Alternative approaches using imaging or other label-free techniques often require highly sophisticated setups, are commonly limited in resolution, and produce challenging amounts of data. Our technology exploits scattered light instead. The custom-built flow cytometry setup comprises a fiber array in forward scatter detection for angular resolution and captures the whole pulse shape with advanced signal processing. Thereby this setup enables cell analysis and sorting purely based on scattered light signals without the need for fluorescent labels. We demonstrate the feasibility of this cell sorting technology by sorting cell lines for their cell cycle stages based on scattered light. Furthermore, we demonstrate the ability to classify human peripheral blood T- and B-cell subsets.
流式细胞术是一种高通量分析和分选细胞或颗粒的关键技术。传统和当前的流式细胞术主要基于荧光染色来检测感兴趣的细胞。然而，这些染色剂也有缺点，因为必须仔细测试它们对细胞的影响，以避免对实验结果产生影响。使用成像或其他无标记技术的替代方法通常需要高度复杂的设置，通常在分辨率上受到限制，并且产生具有挑战性的数据量。我们的技术利用散射光。定制的流式细胞术设置包括用于角分辨率的前向散射检测中的光纤阵列，并通过先进的信号处理捕获整个脉冲形状。因此，该设置能够仅基于散射光信号进行细胞分析和分选，而无需荧光标记。我们通过基于散射光对细胞周期阶段进行分选来证明这种细胞分选技术的可行性。此外，我们还证明了对人外周血 T 细胞和 B 细胞亚群进行分类的能力。
