We present the implementation of a combined digital scanned light-sheet microscope (DSLM) able to work in the linear and nonlinear regimes under either Gaussian or Bessel beam excitation schemes. A complete characterization of the setup is performed and a comparison of the performance of each DSLM imaging modality is presented using in vivoCaenorhabditis elegans samples. We found that the use of Bessel beam nonlinear excitation results in better image contrast over a wider field of view.

This work is supported by the Generalitat de Catalunya grant 2009-SGR-159, the Spanish government grants TEC2009-09698 and SAF2008-00211, the EU project STELUM (FP7-PEOPLE-2007-3-1-IAPP, 217997), the Swiss National Science Foundation (SNSF) Sinergia project grant CRSII3_125447, the FP7-NMP VIBRANT project grant CP-IP 228933-2, the NoE P4L and Laserlab optobio. Jacob Licea-Rodriguez acknowledges CONACYT Mexico for supporting his stay at ICFO through a mixed fellowship. This research has been partially supported by Fundació Cellex Barcelona and was mainly conducted at ICFO’s “Super-Resolution Light Microscopy and Nanoscopy Facility”