title: Rapid spontaneous Raman light sheet microscopy using cw-lasers and tunable filters.
abstract: 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.
authors.affiliation: Centro de Investigación Científica y de Educación Superior de Ensenada, Carretera Ensenada-Tijuana, No. 3918, Zona Playitas, 22860 Ensenada B.C., Mexico ; irocha@cicese.mx.
authors.forename: Israel
authors.initials: I
authors.lastname: Rocha-Mendoza
authors.affiliation: ICFO-Institut de Ciencies Fotoniques, Av. Carl Friedrich Gauss, 3 08860 Castelldefels (Barcelona), Spain.
authors.forename: Jacob
authors.initials: J
authors.lastname: Licea-Rodriguez
authors.affiliation: ICFO-Institut de Ciencies Fotoniques, Av. Carl Friedrich Gauss, 3 08860 Castelldefels (Barcelona), Spain.
authors.forename: Mónica
authors.initials: M
authors.lastname: Marro
authors.affiliation: ICFO-Institut de Ciencies Fotoniques, Av. Carl Friedrich Gauss, 3 08860 Castelldefels (Barcelona), Spain.
authors.forename: Omar E
authors.initials: OE
authors.lastname: Olarte
authors.affiliation: Centro de Investigación Científica y de Educación Superior de Ensenada, Carretera Ensenada-Tijuana, No. 3918, Zona Playitas, 22860 Ensenada B.C., Mexico.
authors.forename: Marcos
authors.initials: M
authors.lastname: Plata-Sanchez
authors.affiliation: ICFO-Institut de Ciencies Fotoniques, Av. Carl Friedrich Gauss, 3 08860 Castelldefels (Barcelona), Spain ; pablo.loza@icfo.es.
authors.forename: Pablo
authors.initials: P
authors.lastname: Loza-Alvarez
keywords: (110.4234) Multispectral and hyperspectral imaging; (170.3880) Medical and biological imaging; (180.5655) Raman microscopy; (300.6450) Spectroscopy, Raman