Label-free Raman observation of cytochrome c dynamics during apoptosis

We performed label-free observation of molecular dynamics in apoptotic cells by Raman microscopy. Dynamic changes in cytochrome c distribution at the Raman band of 750 cm-1 were observed after adding an apoptosis inducer to the cells. The comparison of mitochondria fluorescence images and Raman imag...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 2012-01, Vol.109 (1), p.28-32
Main Authors: Okada, Masaya, Smith, Nicholas Isaac, Palonpon, Almar Flotildes, Endo, Hiromi, Kawata, Satoshi, Sodeoka, Mikiko, Fujita, Katsumasa
Format: Article
Language:English
Subjects:
Apoptosis
Cells
cytochrome c
Cytochromes
Cytochromes c - metabolism
dyes
Fluorescence
HeLa Cells
Humans
Imaging
membrane potential
Membrane Potential, Mitochondrial
Microscopy
Microscopy, Fluorescence
Mitochondria
Mitochondrial membranes
molecular dynamics
Molecules
monitoring
Oxidation-Reduction
Physical Sciences
Raman scattering
Solutions
Spectrum Analysis, Raman
Staining and Labeling
Time-Lapse Imaging
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Summary:We performed label-free observation of molecular dynamics in apoptotic cells by Raman microscopy. Dynamic changes in cytochrome c distribution at the Raman band of 750 cm-1 were observed after adding an apoptosis inducer to the cells. The comparison of mitochondria fluorescence images and Raman images of cytochrome c confirmed that changes in cytochrome c distribution can be distinguished as release of cytochrome c from mitochondria. Our observation also revealed that the redox state of cytochrome c was maintained during the release from the mitochondria. Monitoring mitochondrial membrane potential with JC-1 dye confirmed that the observed cytochrome c release was associated with apoptosis.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.1107524108