High-throughput single-cell sorting by stimulated Raman-activated cell ejection

Journal article

Raman-activated cell sorting isolates single cells in a nondestructive and label-free manner, but its throughput is limited by small spontaneous Raman scattering cross section. Coherent Raman scattering integrated with microfluidics enables high-throughput cell analysis, but faces challenges with small cells (<3 μm) and tissue sections. Here, we report stimulated Raman-activated cell ejection (S-RACE) that enables high-throughput single-cell sorting by integrating stimulated Raman imaging, in situ image decomposition, and laser-induced cell ejection. S-RACE allows ejection of live bacteria or fungi guided by their Raman signatures. Furthermore, S-RACE successfully sorted lipid-rich Rhodotorula glutinis cells from a cell mixture with a throughput of ~13 cells per second, and the sorting results were confirmed by downstream quantitative polymerase chain reaction. Beyond single cells, S-RACE shows high compatibility with tissue sections. Incorporating a closed-loop feedback control circuit further enables real-time SRS imaging-identification-ejection. In summary, S-RACE opens exciting opportunities for diverse single-cell sorting applications.

Authors: Zhang, J; Lin, H; Xu, J; Zhang, M; Ge, X

• Publication status: Published
• Peer review status: Peer reviewed
• Publisher: American Association for the Advancement of Science
• Journal: Science Advances
• Volume: 10
• Issue: 50
• Article number: eadn6373
• Place of publication: United States
• Publication date: 2024-12-11
• Acceptance date: 2024-05-21
• DOI: 10.1126/sciadv.adn6373
• EISSN: 2375-2548
• Pmid: 39661682
• Language: English
• Keywords: high-throughput screening assays; spectrum analysis; Raman; microfluidics; flow cytometry; rhodotorula; cell separation; single-cell analysis