24 March 2005 Three-dimensional optical computed-tomography microscope
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Abstract
We present our recent results on the development of three-dimensional (3-D) optical computed- tomography microscope. The instrument is a novel imaging device for the 3-D visualization and quantitative analysis of absorption-stained biological samples. The first instrument developed by our group at the BC Cancer Research Centre used a digital micromirror device (DMD) as a spatial light modulator to control the angles of illumination. This new embodiment employs an optical scanner instead of the DMD. The optical scanner is placed in the illumination path of the microscope system, conjugate to the field plane. The optical system includes also two high numerical aperture objective lenses, a sample stage, a light source, and a CCD camera. Projections are acquired by illuminating a specimen at a number of selected angles within the numerical aperture of the objective (0 < φ < 135°). A new reconstruction algorithm that employs both transform-based and iterative methods is developed to address the limited-angle reconstruction problem. A transform-based reconstruction is used as an initial starting point for the following iterative reconstruction. A feedback correction of the reconstructed image is made on each iteration step. The algorithm enables to incorporate previously known information about the object into the reconstruction process, and improves the reconstruction accuracy. Microscopic 3-D volume reconstructions of quantitatively absorption-stained cells have been generated. The system enables one to look at multiple optical levels of a specimen, and at more natural tissue architecture, including intact cells. Axial and lateral resolutions were measured to be better than 6 microns.
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Ravil Chamgoulov, Pierre Lane, Calum MacAulay, "Three-dimensional optical computed-tomography microscope", Proc. SPIE 5701, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XII, (24 March 2005); doi: 10.1117/12.591859; https://doi.org/10.1117/12.591859
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