| Autor: |
Brusnichkin, Anton V., Nedosekin, Dmitry A., Galanzha, Ekaterina I., Vladimirov, Yuri A., Shevtsova, Elena F., Proskurnin, Mikhail A., Zharov, Vladimir P. |
| Zdroj: |
Journal of Biophotonics; Dec2010, Vol. 3 Issue 12, p791-806, 16p |
| Abstrakt: |
Light-absorbing endogenous cellular proteins, in particular cytochrome c, are used as intrinsic biomarkers for studies of cell biology and environment impacts. To sense cytochrome c against real biological backgrounds, we combined photothermal (PT) thermal-lens single-channel schematic in a back-synchronized measurement mode and a multiplex thermal-lens schematic in a transient high resolution ( ca. 350 nm) imaging mode. These multifunctional PT techniques using continuous-wave (cw) Ar laser and a nanosecond pulsed optical parametric oscillator in the visible range demonstrated the capability for label-free spectral identification and quantification of trace amounts of cytochrome c in a single mitochondrion alone or within a single live cell. PT imaging data were verified in parallel by molecular targeting and fluorescent imaging of cellular cytochrome c. The detection limit of cytochrome c in a cw mode was 5 × 10 mol/L (80 attomols in the signal-generation zone); that is ca. 10lower than conventional absorption spectroscopy. Pulsed fast PT microscopy provided the detection limit for cytochrome c at the level of 13 zmol (13 × 10 mol) in the ultrasmall irradiated volumes limited by optical diffraction effects. For the first time, we demonstrate a combination of high resolution PT imaging with PT spectral identification and ultrasensitive quantitative PT characterization of cytochrome c within individual mitochondria in single live cells. A potential of far-field PT microscopy to sub-zeptomol detection thresholds, resolution beyond diffraction limit, PT Raman spectroscopy, and 3D imaging are further highlighted. (© 2010 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim) [ABSTRACT FROM AUTHOR] |
| Databáze: |
Complementary Index |
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