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J Biomed Opt. 2008 Jul-Aug;13(4):044027. doi: 10.1117/1.2952192.

Raman microscopy for dynamic molecular imaging of living cells.

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Osaka University, Department of Frontier Biosciences, 1-3 Yamadaoka, Suita, Osaka 565-0871, Japan.


We demonstrate dynamic imaging of molecular distribution in unstained living cells using Raman scattering. By combining slit-scanning detection and optimizing the excitation wavelength, we imaged the dynamic molecular distributions of cytochrome c, protein beta sheets, and lipids in unstained HeLa cells with a temporal resolution of 3 minutes. We found that 532-nm excitation can be used to generate strong Raman scattering signals and to suppress autofluorescence that typically obscures Raman signals. With this technique, we reveal time-resolved distributions of cytochrome c and other biomolecules in living cells in the process of cytokinesis without the need for fluorescent labels or markers.

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