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Optical projection tomography (OPT) is the technique used to generate the 3D models we display on FishNet. We take a series of 2D images and using these reconstruct the 3D organization of the sample. One of the simplest ways of doing this is using back projection. Light is shone through a sample creating a projection image as shown below. The amount of light passing through the sample is dependent on the thickness of the sample, where it is thicker less light passes through the sample. The sample is rotated and the process is repeated. The shape of the original object can be reconstructed using a simple back projection method overlaying the projection images as shown.
The same principle is used in OPT, however 400 images are captured over 360 O
and an algebraic method of reconstruction replaces the simple overlapping and eliminates the background star-shaped pattern.
Reconstruction of each section results in a complete three-dimensional (3D) model.
The process of Optical Projection Tomography (OPT) was described by Sharpe et al. 2002 and is described on the Sharpe lab webpage, questions concerning this technology should be directed to them. The application of OPT to fish was developed in the Currie lab and is described in Bryson-Richardson and Currie (Bryson-Richardson and Currie 2004). References
Bryson-Richardson RJ, Currie PD.Optical projection tomography for spatio-temporal analysis in the zebrafish.
Methods Cell Biol. 2004;76:37-50
Sharpe J, Ahlgren U, Perry P, Hill B, Ross A, Hecksher-Sorensen J, Baldock
R, Davidson D. Optical projection tomography as a tool for 3D microscopy and gene expression studies.
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