据研究人员介绍，对具有独特空间和形态特征的单细胞进行快速和选择性的分离仍然是一项技术挑战。
研究人员通过建立高速图像驱动的细胞分拣（ICS）来解决了这个问题，它可以记录多色荧光图像，并根据图像数据的测量结果以每秒15,000次的速度分拣细胞。结果表明，ICS可以量化细胞形态和标记蛋白的定位，并通过分离有丝分裂阶段提高细胞周期分析的分辨率。研究人员将ICS与CRISPR-pooled筛选相结合来确定了核因子κB（NF-κB）途径的调节因子，并实现全基因组图像筛选在大约9小时的运行时间内完成。通过评估复杂的细胞表型，ICS大大扩展了细胞分选应用和集合遗传筛选所能获得的表型空间。
附：英文原文
Title: High-speed fluorescence image–enabled cell sorting
Author: Daniel Schraivogel, Terra M. Kuhn, Benedikt Rauscher, Marta Rodríguez-Martínez, Malte Paulsen, Keegan Owsley, Aaron Middlebrook, Christian Tischer, Beáta Ramasz, Diana Ordoez-Rueda, Martina Dees, Sara Cuylen-Haering, Eric Diebold, Lars M. Steinmetz
Issue&Volume: 2022-01-21
Abstract: 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.
DOI: abj3013
Source: https://www.science.org/doi/10.1126/science.abj3013