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Vibrationally resonant imaging of a single living cell by supercontinuum-based multiplex coherent anti-Stokes Raman scattering microspectroscopy

Hideaki Kano and Hiro-o Hamaguchi

Open Access Open Access

Abstract

Supercontinuum-based multiplex coherent anti-Stokes Raman scattering (CARS) microspectroscopy has been applied to vibrational imaging of a living fission yeast cell. We have successfully extracted only a vibrationally resonant CARS image from a characteristic spectral profile in the C-H stretching vibrational region. Using our simple but sensitive analysis, the vibrational contrast is significantly improved in comparison with a CARS imaging at a fixed Raman shift. The CARS image of a living yeast cell indicates several areas at which the signal is remarkably strong. They are considered to arise from mitochondria.

©2005 Optical Society of America

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Figures (3)
Fig. 1.
Fig. 1. Typical spectral profile of the CARS signal of a living yeast cell (red solid) and surrounding water (blue dotted)

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Fig. 2.
Fig. 2. CARS lateral images of living yeast cells at the Raman shift of 2856 cm-1 (a) and 2200 cm-1 (b). The black and white crosses in (a) correspond to the positions at which the spectral profiles are shown in Fig. 1.

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Fig. 3.
Fig. 3. Vibrationally resonant CARS imaging using the differentiation method (a) and a method using a conventional lineshape function described in Eq. (1) (b). Note that the contrast and signal-to-noise ratio is dramatically improved in comparison with Fig. 2(a).

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Equations (1)

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I ( ω ) = ∣ A NR e i ϕ + ∑ R A R Γ R Γ R − i ( ω − Ω R ) ∣ 2 .
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