Journal
BIOMEDICAL OPTICS EXPRESS
Volume 6, Issue 9, Pages 3449-3461Publisher
OPTICAL SOC AMER
DOI: 10.1364/BOE.6.003449
Keywords
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Funding
- National Research and Technology Council of Mexico (CONACyT) [233012]
- CONACyT [155803, 238390]
- Laser Lab Europe (EC's Seventh Framework Programme) [284464]
- Photonics4Life network of excellence
- European regional Development Fund
- Fundacio Cellex Barcelona
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We perform rapid spontaneous Raman 2D imaging in light-sheet microscopy using continuous wave lasers and interferometric tunable filters. By angularly tuning the filter, the cut-on/off edge transitions are scanned along the excited Stokes wavelengths. This allows obtaining cumulative intensity profiles of the scanned vibrational bands, which are recorded on image stacks; resembling a spectral version of the knife-edge technique to measure intensity profiles. A further differentiation of the stack retrieves the Raman spectra at each pixel of the image which inherits the 3D resolution of the host light sheet system. We demonstrate this technique using solvent solutions and composites of polystyrene beads and lipid droplets immersed in agar and by imaging the C-H (2800-3100 cm(-1)) region in a C. elegans worm. The image acquisition time results in 4 orders of magnitude faster than confocal point scanning Raman systems, allowing the possibility of performing fast spontaneous Raman.3D-imaging on biological samples. (C) 2015 Optical Society of America
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