Hyperspectral stimulated Raman scattering (SRS) microscopy allows imaging of complex chemical mixtures and analysis cellular metabolites with high specificity. However, current SRS imaging is not implemented to address the cell heterogeneity issue, which can only be resolved by statistical analysis of a large amount of cells through cytometry. We developed a high-speed hyperspectral SRS image cytometry platform based on multiplex excitation, acquiring a Raman spectrum of 200 wavenumbers in 5 microseconds. This platform enables measurement of <100 cells per second. Multiple chemical signatures, featuring different cellular organelles such as lipids, endoplasmic reticulum, nucleus, and cytoplasm can be segmented. Statistical analysis over a large amount of cells reveals unprecedented details about cell metabolic changes after drug treatment.

Date Published: 22 February 2019
PDF: 8 pages
Proc. SPIE 10882, Multiphoton Microscopy in the Biomedical Sciences XIX, 108822E (22 February 2019); doi: 10.1117/12.2510871

Published in SPIE Proceedings Vol. 10882:
Multiphoton Microscopy in the Biomedical Sciences XIX
Ammasi Periasamy; Peter T. C. So; Karsten König, Editor(s)