Open AccessRapid spontaneous Raman light sheet microscopy using cw-lasers and tunable filters
Author(s) -
Israel RochaMendoza,
Jacob Licea-Rodríguez,
Mónica Marro,
Omar E. Olarte,
Marcos Plata-Sanchez,
Pablo Loza-Álvarez
Publication year - 2015
Publication title -
biomedical optics express
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.362
H-Index - 86
ISSN - 2156-7085
DOI - 10.1364/boe.6.003449
Subject(s) - raman spectroscopy , materials science , optics , microscopy , laser , imaging spectroscopy , microscope , wavelength , light sheet fluorescence microscopy , optoelectronics , spectroscopy , scanning confocal electron microscopy , physics , quantum mechanics
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 (2800-3100cm(-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.