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Proceedings Paper

Dual-CARS microscopy
Author(s): Annika M. Enejder; Christian Brackmann; Ondrej Burkacky; Madeleine Åkeson
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Paper Abstract

We present a new Coherent Anti-Stokes Raman Scattering (CARS) microscopy technique for label-free imaging of biomolecules in living cells; dual-CARS microscopy. The use of three synchronized laser pulses in a dual-pump/dualdetection configuration enables imaging of two species with different molecular vibrations simultaneously, as well as acquisition of images free of non-resonant background. We show the power of the method by imaging deuterated nonadecane slowly diffusing into a suspension of living yeast cells in medium, clearly distinguishing the medium and the lipid droplets in the cells by probing the CH2 vibration from the D-nonadecane by probing the CD vibration. In addition, images of lipid stores in living C. elegans nematodes free of non-resonant background are shown. This results in a significant enhancement of the image contrast, allowing the visualization of emerging, low-density lipid stores in a dauer larva, difficult to distinguish in conventional CARS microscopy. The separation of the non-resonant background is shown to be beneficial also when monitoring molecules with weak vibrational modes. The improved sensitivity obtained is illustrated by probing the C=C vibration in polyunsaturated lipids extracted from fish. This enables the monitoring of the degree of unsaturation of lipids, a high value of which is reported in foods known to have positive effects on human health.

Paper Details

Date Published: 10 February 2007
PDF: 8 pages
Proc. SPIE 6442, Multiphoton Microscopy in the Biomedical Sciences VII, 64420G (10 February 2007); doi: 10.1117/12.714388
Show Author Affiliations
Annika M. Enejder, Chalmers Univ. of Technology (Sweden)
Christian Brackmann, Chalmers Univ. of Technology (Sweden)
Ondrej Burkacky, Chalmers Univ. of Technology (Sweden)
Madeleine Åkeson, Chalmers Univ. of Technology (Sweden)

Published in SPIE Proceedings Vol. 6442:
Multiphoton Microscopy in the Biomedical Sciences VII
Ammasi Periasamy; Peter T. C. So, Editor(s)

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