Genome Biology Unit, European Molecular Biology Laboratory (EMBL), Heidelberg, Germany.
Cell Biology and Biophysics Unit, EMBL, Heidelberg, Germany.
Science. 2022 Jan 21;375(6578):315-320. doi: 10.1126/science.abj3013. Epub 2022 Jan 20.
Fast and selective isolation of single cells with unique spatial and morphological traits remains a technical challenge. Here, we address this by establishing high-speed image-enabled cell sorting (ICS), which records multicolor fluorescence images and sorts cells based on measurements from image data at speeds up to 15,000 events per second. We show that ICS quantifies cell morphology and localization of labeled proteins and increases the resolution of cell cycle analyses by separating mitotic stages. We combine ICS with CRISPR-pooled screens to identify regulators of the nuclear factor κB (NF-κB) pathway, enabling the completion of genome-wide image-based screens in about 9 hours of run time. By assessing complex cellular phenotypes, ICS substantially expands the phenotypic space accessible to cell-sorting applications and pooled genetic screening.
快速而选择性地分离具有独特空间和形态特征的单细胞仍然是一项技术挑战。在这里，我们通过建立高速图像引导的细胞分选（ICS）来解决这个问题，该方法可以记录多色荧光图像，并根据图像数据的测量结果以高达每秒 15,000 个事件的速度对细胞进行分选。我们表明，ICS 可以定量细胞形态和标记蛋白的定位，并通过分离有丝分裂阶段来提高细胞周期分析的分辨率。我们将 ICS 与 CRISPR 池筛选相结合，以鉴定核因子 κB（NF-κB）途径的调节剂，从而能够在大约 9 小时的运行时间内完成基于全基因组的图像筛选。通过评估复杂的细胞表型，ICS 大大扩展了可用于细胞分选应用和基因池筛选的表型空间。
