3D Microscopy by orbital radially modulated scan: Fundaments and applications.

MANUELA GABRIEL; LAURA C ESTRADA; MARÍA CECILIA ZAZA

Pucón, Región de la Araucanía

Conferencia; RIAO/OPTILAS-IX Iberoamerican meeting on optics and XII Iberoamerican meeting on optics, lasers and applications; 2016

Centro de Óptica y Fotónica de la Universidad de Concepción (CEFOP-UDEC)

In this work, we present an alternative imaging method based on the orbital scan of a laser excitation beam around the object of interest. This technology is capable of producing tridimensional images of fluorescent structures with nanometrical resolution in a few milliseconds. The method relies on the fact that when the excitation beam is near a fluorescent object, the emitted light from the object depends on the distance between its surface and the excitation beam. Modulating the distance between the beam and the object and taking into account the nonlinearity of the excitation intensity profile, it is possible to obtain an oscillating response whose amplitude depends only on the distance to the surface of the object. Given the fact that the excitation beam is always near the structure of interest, it is possible to measure moving specimens.Here, we present numerical simulations that demonstrate the method performance, as well as control experiments on tridimensional fluorescent samples fabricated by optical lithography. We show tridimensional reconstructions of Arabipdosis Thaliana roots which are ~ 50 um in lengths and ~ 5 um in diameters. Finally, we show that we can replace the traditional focus system, based on the mechanical movement of the objective lens by a simple new system with an electrically tunable lens, with a velocity 1 fold increase and a scan range 2 fold increase.