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