High-speed fluorescence image-enabled cell sorting

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 kappa B (NF-kappa B) 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.

Automated summary

In this study, a high-speed image-enabled cell sorting method was established to isolate single cells with unique spatial and morphological traits. This method allows for quantitative analysis of cell morphology and localization of labeled proteins, and improves the resolution of cell cycle analysis. By combining it with CRISPR-pooled screens, comprehensive genome-wide image-based screens can be completed in a short period of time.