Dual‐polarization Raman spectral imaging to extract overlapping molecular fingerprints of living cells

Abstract

Raman spectral imaging is gaining more and more attention in biological studies because of its label-free characteristic. However, the discrimination of overlapping chemical contrasts has been a major challenge. In this study, we introduce an optical method to simultaneously obtain two orthogonally polarized Raman images from a single scan of the sample. We demonstrate how this technique can improve the quality and quantity of the hyperspectral Raman dataset and how the technique is expected to further extend the horizons of Raman spectral imaging in biological studies by providing more detailed chemical information. The dual-polarization Raman images of a HeLa cell.

Protein expression guided chemical profiling of living cells by the simultaneous observation of Raman scattering and anti-Stokes fluorescence emission

“…The cytochrome c , protein and lipid Raman images in Fig. 3 all correspond well to our previously reported Raman images of the corresponding vibrational modes3435, with minimal cross-talk from the anti-Stokes fluorescence image of the same hyperspectral dataset. This indicates that our hybrid fluorescence-Raman imaging technique indeed provides authentic fluorescence and Raman information for further analysis.…”

“…Both the HeLa and ES cell lines are the same ones used in our previous researches and cultured in the same condition434. The cell lines have not been authenticated and tested for mycoplasma contamination during the period of this study.…”

Label‐Free Molecular Imaging of Biological Cells and Tissues by Linear and Nonlinear Raman Spectroscopic Approaches

“…[77] Twoo rthogonally polarized Raman images were simultaneously obtained from asingle scan of the sample.T his technique can improve the quality,q uantity,a nd contrast of hyperspectral datasets, which was demonstrated for living cells. [77] Twoo rthogonally polarized Raman images were simultaneously obtained from asingle scan of the sample.T his technique can improve the quality,q uantity,a nd contrast of hyperspectral datasets, which was demonstrated for living cells.…”

Modern trends in biophotonics for clinical diagnosis and therapy to solve unmet clinical needs

“…An optical method was introduced to simultaneously obtain two orthogonally polarized Raman images from a single scan . The authors demonstrated on single living cells how the technique improves the quality and quantity of hyperspectral data sets (see Figure ).…”