Abstract

3D microscopy by orbital radially modulated scan. An. AFA [online]. 2018, vol.29, n.1, pp.12-19. ISSN 0327-358X.

In this work, we present an alternative imaging method based on the orbital scanning 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. By 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 tridimensional reconstructions of Arabidopsis Thaliana roots which are ~ 50 μm in length and ~ 5 μm in diameter.

Keywords : microscopy; fluorescence; single particle tracking.

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