Journal Article

Label-free cell cycle analysis for high-throughput imaging flow cytometry

Summers, Huw D., Cerveira, Joana, Patterson, James O., Filby, Andrew, Blasi, Thomas, Hennig, Holger, Rees, Paul, Davies, Derek, Carpenter, Anne E., Theis, Fabian J.

2016

Overview

Imaging flow cytometry combines the high-throughput capabilities of conventional flow cytometry with single-cell imaging. Here we demonstrate label-free prediction of DNA content and quantification of the mitotic cell cycle phases by applying supervised machine learning to morphological features extracted from brightfield and the typically ignored darkfield images of cells from an imaging flow cytometer. This method facilitates non-destructive monitoring of cells avoiding potentially confounding effects of fluorescent stains while maximizing available fluorescence channels. The method is effective in cell cycle analysis for mammalian cells, both fixed and live, and accurately assesses the impact of a cell cycle mitotic phase blocking agent. As the same method is effective in predicting the DNA content of fission yeast, it is likely to have a broad application to other cell types. Imaging flow cytometry enables high-throughput acquisition of fluorescence, brightfield and darkfield images of biological cells. Here, Blasi et al. demonstrate that applying machine learning algorithms on brightfield and darkfield images can detect cellular phenotypes without the need for fluorescent stains, enabling label-free assays.

• Publisher: Nature Publishing Group UK,Nature Publishing Group,Nature Portfolio
• Subject: 14 / 631/1647/1407/1492 / 631/1647/48 / 631/80/641 / Cell Cycle - physiology / Deoxyribonucleic acid / DNA / DNA - genetics / Flow Cytometry - methods / Humanities and Social Sciences / Humans / Image Processing, Computer-Assisted / Jurkat Cells / Machine Learning / multidisciplinary / Schizosaccharomyces / Science / Science (multidisciplinary) / Yeasts