title: Raman microscopy for dynamic molecular imaging of living cells.
abstract: 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.
authors.affiliation: Osaka University, Department of Frontier Biosciences, 1-3 Yamadaoka, Suita, Osaka 565-0871, Japan.
authors.forename: Keisaku
authors.initials: K
authors.lastname: Hamada
authors.forename: Katsumasa
authors.initials: K
authors.lastname: Fujita
authors.forename: Nicholas Isaac
authors.initials: NI
authors.lastname: Smith
authors.forename: Minoru
authors.initials: M
authors.lastname: Kobayashi
authors.forename: Yasushi
authors.initials: Y
authors.lastname: Inouye
authors.forename: Satoshi
authors.initials: S
authors.lastname: Kawata
mesh_terms: D001704:Biopolymers / Q000032:analysis*; D006367:HeLa Cells; D006801:Humans; D008853:Microscopy / Q000379:methods*; D015336:Molecular Probe Techniques; D013059:Spectrum Analysis, Raman / Q000379:methods*
chemical_list: D001704:Biopolymers