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Proceedings Paper

In vivo imaging of zebrafish from embryo to adult stage with optical projection tomography
Author(s): Andrea Bassi; Luca Fieramonti; Cosimo D'Andrea; Gianluca Valentini; Rinaldo Cubeddu; Sandro De Silvestri; Giulio Cerullo; Efrem Foglia; Franco Cotelli
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Paper Abstract

Optical Projection Tomography (OPT) is a three dimensional imaging technique that is particularly suitable for studying millimeter sized biological samples and organisms. Similarly to x-ray computed tomography, OPT is based on the acquisition of a sequence of images taken through the sample at many angles (projections). Assuming the linearity of the optical absorption process, the projections are combined to reconstruct the 3-D volume of the sample, typically using a filtered back-projection algorithm. OPT has been applied to in-vivo imaging of zebrafish (Danio rerio). The instrument and the protocol for in vivo imaging of zebrafish embryos and juvenile specimens are described. Light scattering remains a challenge for in vivo OPT, especially when samples at the upper size limit, like zebrafish at the adult stage, are under study. We describe Time-Gated Optical Projection Tomography (TGOPT), a technique able to reconstruct adult zebrafish internal structures by counteracting the scattering effects through a fast time-gate. The time gating mechanism is based on non-linear optical upconversion of an infrared ultrashort laser pulse and allows the detection of quasi-ballistic photons within a 100 fs temporal gate. This results in a strong improvement in contrast and resolution with respect to conventional OPT. Artifacts in the reconstructed images are reduced as well. We show that TGOPT is suited for imaging the skeletal system and nervous structures of adult zebrafish.

Paper Details

Date Published: 26 March 2013
PDF: 7 pages
Proc. SPIE 8593, Optical Methods in Developmental Biology, 85930A (26 March 2013); doi: 10.1117/12.2004340
Show Author Affiliations
Andrea Bassi, Politecnico di Milano (Italy)
Luca Fieramonti, Politecnico di Milano (Italy)
Cosimo D'Andrea, Politecnico di Milano (Italy)
Gianluca Valentini, Politecnico di Milano (Italy)
Rinaldo Cubeddu, Politecnico di Milano (Italy)
Sandro De Silvestri, Politecnico di Milano (Italy)
Giulio Cerullo, Politecnico di Milano (Italy)
Efrem Foglia, Univ. degli Studi di Milano (Italy)
Franco Cotelli, Univ. degli Studi di Milano (Italy)

Published in SPIE Proceedings Vol. 8593:
Optical Methods in Developmental Biology
Andrew M. Rollins; Cecilia Lo; Scott E. Fraser, Editor(s)

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