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Cellular Oncology
Volume 26 (2004), Issue 5-6, Pages 319-327
http://dx.doi.org/10.1155/2004/209579

Optical Computed-Tomographic Microscope for Three-Dimensional Quantitative Histology

Ravil Chamgoulov, Pierre Lane, and Calum MacAulay

Cancer Imaging Department, BC Cancer Research Centre, Vancouver, Canada

Copyright © 2004 Hindawi Publishing Corporation and the authors. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

A novel optical computed‐tomographic microscope has been developed allowing quantitative three‐dimensional (3D) imaging and analysis of fixed pathological material. Rather than a conventional two‐dimensional (2D) image, the instrument produces a 3D representation of fixed absorption‐stained material, from which quantitative histopathological features can be measured more accurately. The accurate quantification of these features is critically important in disease diagnosis and the clinical classification of cancer. The system consists of two high NA objective lenses, a light source, a digital spatial light modulator (DMD, by Texas Instrument), an x–y stage, and a CCD detector. The DMD, positioned at the back pupil‐plane of the illumination objective, is employed to illuminate the specimen with parallel rays at any desired angle. The system uses a modification of the convolution backprojection algorithm for reconstruction. In contrast to fluorescent images acquired by a confocal microscope, this instrument produces 3D images of absorption stained material. Microscopic 3D volume reconstructions of absorption‐stained cells have been demonstrated. Reconstructed 3D images of individual cells and tissue can be cut virtually with the distance between the axial slices less than 0.5 μm.