Holzner Gregor, Mateescu Bogdan, van Leeuwen Daniel, Cereghetti Gea, Dechant Reinhard, Stavrakis Stavros, deMello Andrew
Institute for Chemical & Bioengineering, ETH Zürich, Vladimir Prelog Weg 1, 8093 Zürich, Switzerland.
Brain Research Institute, University of Zurich, Winterthurerstrasse 190, 8057 Zurich, Switzerland.
Cell Rep. 2021 Mar 9;34(10):108824. doi: 10.1016/j.celrep.2021.108824.
We present a sheathless, microfluidic imaging flow cytometer that incorporates stroboscopic illumination for blur-free fluorescence detection at ultra-high analytical throughput. The imaging platform is capable of multiparametric fluorescence quantification and sub-cellular localization of these structures down to 500 nm with microscopy image quality. We demonstrate the efficacy of the approach through the analysis and localization of P-bodies and stress granules in yeast and human cells using fluorescence and bright-field detection at analytical throughputs in excess of 60,000 and 400,000 cells/s, respectively. Results highlight the utility of our imaging flow cytometer in directly investigating phase-separated compartments within cellular environments and screening rare events at the sub-cellular level for a range of diagnostic applications.
我们展示了一种无鞘微流控成像流式细胞仪，该仪器采用频闪照明技术，可在超高分析通量下实现无模糊荧光检测。该成像平台能够进行多参数荧光定量分析，并能以显微镜图像质量对这些结构进行亚细胞定位，定位精度可达500纳米。我们通过分别在超过60000个细胞/秒和400000个细胞/秒的分析通量下，利用荧光和明场检测对酵母和人类细胞中的P小体和应激颗粒进行分析和定位，证明了该方法的有效性。结果突出了我们的成像流式细胞仪在直接研究细胞环境中相分离区室以及在亚细胞水平筛选罕见事件以用于一系列诊断应用方面的实用性。
