Abstract
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 vivo Caenorhabditis elegans samples. We found that the use of Bessel beam nonlinear excitation results in better image contrast over a wider field of view.
Highlights
Traditional wide-field optical fluorescence microscopes are proven invaluable tools that accomplish the most diverse imaging tasks at the cellular and sub-cellular level
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
We found that the use of Bessel beam nonlinear excitation results in better image contrast over a wider field of view
Summary
Traditional wide-field optical fluorescence microscopes are proven invaluable tools that accomplish the most diverse imaging tasks at the cellular and sub-cellular level. That is why having an alternative technique that would allow the observation of fast events with high spatial resolutions over a large field of view (FOV) is extremely important. To overcome the problem of out-of-focus light, techniques referred as laser point scanning microscopy (LSM), such as confocal and multi-photon microscopies, have been introduced [1]. LSM techniques generate images only from in-focus light providing intrinsic optical sectioning. In addition to out-of-focus light, another important issue to take into consideration when imaging biological samples is the photodamage (photobleaching and phototoxicity). In LSM techniques, as excitation and collection occurs along the same axis, the entire sample is repeatedly irradiated when taking an image stack. Cumulative photodamage is induced within the sample [2]
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
