<title>Fluorescence microtomography: multiangle image acquisition and 3D digital reconstruction</title>

Abstract

We have developed a prototype fluorescence microscope which, using tomographic image acquisition and reconstruction techniques, can automatically combine conventional and/or confocal image stacks taken at a number of orientations into a single, very-high-resolution 3D image. We use the term `microtomography' in a broad sense to denote digital image reconstruction from multiple imaging operations which are not necessarily projections. Our system holds a biological specimen inside a thin capillary tube which is rotatable over a 360 degree range beneath an immersion objective. 3D fluorescence image data volumes are acquired by frame-grabbing a through-focus series of 2D images at each angle of rotation. Digital reconstruction of the multi-angle data volumes produces a single very-high-resolution 3D image and involves algorithms which perform rotation, interpolation, alignment and normalization operations in frequency (Fourier) space.