Comparison of image reconstruction techniques for optical projection tomography.

We present a comparison of image reconstruction techniques for optical projection tomography. We compare conventional filtered back projection, sinogram filtering using the frequency-distance relationship (FDR), image deconvolution, and 2D point-spread-function-based iterative reconstruction. The latter three methods aim to remove the spatial blurring in the reconstructed image originating from the limited depth of field caused by the point spread function of the imaging system. The methods are compared based on simulated data, experimental optical projection tomography data of single fluorescent beads, and high-resolution optical projection tomography imaging of an entire zebrafish larva. We demonstrate that the FDR method performs poorly on data acquired with high numerical aperture optical imaging systems. We show that the deconvolution technique performs best on highly sparse data with low signal-to-noise ratio. The point-spread-function-based reconstruction method is superior for nonsparse objects and data of high signal-to-noise ratio.

Point spread function based image reconstruction in optical projection tomography.

As a result of the shallow depth of focus of the optical imaging system, the use of standard filtered back projection in optical projection tomography causes space-variant tangential blurring that increases with the distance to the rotation axis. We present a novel optical tomographic image reconstruction technique that incorporates the point spread function of the imaging lens in an iterative reconstruction. The technique is demonstrated using numerical simulations, tested on experimental optical projection tomography data of single fluorescent beads, and applied to high-resolution emission optical projection tomography imaging of an entire zebrafish larva. Compared to filtered back projection our results show greatly reduced radial and tangential blurring over the entire [Formula: see text] mm2 field of view, and a significantly improved signal to noise ratio.

Image resolution and deconvolution in optical tomography.

We present a frequency domain analysis of the image resolution of optical tomography systems. The result of our analysis is a description of the spatially-variant resolution in optical tomographic image after reconstruction as a function of the properties of the imaging system geometry. We validate our model using optical projection tomography (OPT) measurements of fluorescent beads embedded in agarose gel. Our model correctly describes both the radial and tangential resolution of the measured images. In addition, we present a correction of the tomographic images for the spatially-varying resolution using a deconvolution algorithm. The resulting corrected tomographic reconstruction shows a homogeneous and isotropic pixel-limited resolution across the entire image. Our method is applied to OPT measurements of a zebrafish, showing improved resolution. Aside from allowing image correction and providing a resolution measure for OPT systems, our model provides a powerful tool for the design of optical tomographic systems.