Research Papers

Raman microscopy for dynamic molecular imaging of living cells

[+] Author Affiliations
Keisaku Hamada

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

Katsumasa Fujita

Osaka University, Department of Applied Physics, 2-1 Yamadaoka, Suita, Osaka 565-0871, Japan and RIKEN, Nanophotonics Laboratory, Hirosawa, Wako, Saitama 351-0198, Japan

Nicholas Isaac Smith

Osaka University, Department of Applied Physics, 2-1 Yamadaoka, Suita, Osaka 565-0871, Japan

Minoru Kobayashi

Nanophoton Corporation, A-509, CASI,Osaka University, 2-1 Yamadaoka, Suita, Osaka 565-0871, Japan

Yasushi Inouye

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

Satoshi Kawata

Osaka University, Department of Applied Physics, 2-1 Yamadaoka, Suita, Osaka 565-0871, Japan and RIKEN, Nanophotonics Laboratory, Hirosawa, Wako, Saitama 351-0198, Japan

J. Biomed. Opt. 13(4), 044027 (July 18, 2008). doi:10.1117/1.2952192
History: Received December 29, 2007; Accepted February 09, 2008; Published July 18, 2008
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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 3minutes. 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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© 2008 Society of Photo-Optical Instrumentation Engineers

Citation

Keisaku Hamada ; Katsumasa Fujita ; Nicholas Isaac Smith ; Minoru Kobayashi ; Yasushi Inouye, et al.
"Raman microscopy for dynamic molecular imaging of living cells", J. Biomed. Opt. 13(4), 044027 (July 18, 2008). ; http://dx.doi.org/10.1117/1.2952192


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