Live single-cell laser tag.

Loïc Binan,Loïc Binan,Louis-Etienne Lorenzo,El Bachir Affar,El Bachir Affar,Jiannis Ragoussis,Jiannis Ragoussis,Jiannis Ragoussis,Karine Choquet,Santiago Costantino,Santiago Costantino,Yu Chang Wang,Yves De Koninck,Yves De Koninck,Karine Choquet,Claudia L Kleinman,Claudia L Kleinman,Javier Mazzaferri

doi:10.1038/ncomms11636

Abstract

The ability to conduct image-based, non-invasive cell tagging, independent of genetic engineering, is key to cell biology applications. Here we introduce cell labelling via photobleaching (CLaP), a method that enables instant, specific tagging of individual cells based on a wide array of criteria such as shape, behaviour or positional information. CLaP uses laser illumination to crosslink biotin onto the plasma membrane, coupled with streptavidin conjugates to label individual cells for genomic, cell-tracking, flow cytometry or ultra-microscopy applications. We show that the incorporated mark is stable, non-toxic, retained for several days, and transferred by cell division but not to adjacent cells in culture. To demonstrate the potential of CLaP for genomic applications, we combine CLaP with microfluidics-based single-cell capture followed by transcriptome-wide next-generation sequencing. Finally, we show that CLaP can also be exploited for inducing transient cell adhesion to substrates for microengineering cultures with spatially patterned cell types.

Highlights

Cellular labels are essential components in the toolbox to build our current understanding of biological function
Specific cells can be chosen based on their morphological characteristics, dynamic behaviour, localization in the sample at a given time, or any visible feature that distinguishes the cells of interest from an ensemble
cell labelling via photobleaching (CLaP) allows combining the accuracy and versatility of image-based selection with the high throughput of automated cell-sorting methods, permitting experiments that account for cellular context or temporal dynamics, such as transcriptomic profiling preserving spatial information

Summary

Introduction

Cellular labels are essential components in the toolbox to build our current understanding of biological function. By choosing among different types of such streptavidin conjugates, cells can be tagged with fluorescence (Fig. 1b–e), electron-dense molecules (Fig. 1f and Supplementary Fig. 1) or other labels. Cells tagged with biotin, resuspended from the substrate with trypsin and reseeded were revealed with fluorescent streptavidin and identified among a large population of unstained cells after 24 h (Supplementary Fig. 2).

Results

Conclusion