Light-sheet fluorescence microscopy is quickly becoming one of the cornerstone imaging techniques in biology as it provides rapid, three-dimensional sectioning of specimens at minimal levels of phototoxicity. It is very appealing to bring this unique combination of imaging properties into an endoscopic setting and be able to perform optical sectioning deep in tissues.
Current endoscopic approaches for delivery of light-sheet illumination are based on single-mode optical fibre terminated by cylindrical gradient index lens. Such configuration generates a light-sheet plane that is axially fixed and a mechanical movement of either the sample or the endoscope is required to acquire three-dimensional information about the sample. Furthermore, the axial resolution of this technique is limited to 5um.
The delivery of the light-sheet through the multimode fibre provides better axial resolution limited only by its numerical aperture, the light-sheet is scanned holographically without any mechanical movement, and multiple advanced light-sheet imaging modalities, such as Bessel and structured illumination Bessel beam, are intrinsically supported by the system due to the cylindrical symmetry of the fibre.
We discuss the holographic techniques for generation of multiple light-sheet types and demonstrate the imaging on a sample of fluorescent beads fixed in agarose gel, as well as on a biological sample of Spirobranchus Lamarcki.
Martin Plöschner, Véra Kollárová, Zbyněk Dostál, Jonathan Nylk, Thomas Barton-Owen, David E. K. Ferrier, Radim Chmelik, Kishan Dholakia, and Tomáš Cizmár, "Generation of light-sheet at the end of multimode fibre (Conference Presentation)," Proc. SPIE 10073, Adaptive Optics and Wavefront Control for Biological Systems III, 1007305 (Presented at SPIE BiOS: January 28, 2017; Published: 24 April 2017); https://doi.org/10.1117/12.2250694.5380600113001.
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Study of self-shadowing effect as a simple means to realize nanostructured thin films and layers with special attentions to birefringent obliquely deposited thin films and photo-luminescent porous silicon