Construction On Optical Projection Tomography System And Research On System Application

Optical Projection Tomography (OPT) is a novel imaging tool which making use of uniform collimated beam transmits through sample, acquiring multi-angle projections or fluorography, then, obtaining three-dimensional (3D) information by reconstruction. OPT can provide both anatomical imaging and molecular specific imaging for mesoscopic specimens with size from about1mm to10mm across. It can conduct qualitative and quantitative research at the cellular level for small scale animals. Meanwhile, It has many advantages such as high resolution, dynamic, no radiation, low cost, easy to use and so on which have met the research requirement in biomedical. However, domestic research on OPT is in its infancy. To keep up with the international pace and promote the OPT application in domestic, we need to study OPT techniques including system construction and imaging method.In this thesis, OPT imaging techniques have been developed, including system construction, research on system application. The main contents of this thesis are listed as follows:In system construction, we have designed and built an OPT imaging system, including hardware and software. Our OPT system constructed by light source model, sample positioning model, image acquisition model and control model. Meanwhile, we have programmed data acquisition software and imaging software. Now, this system has been applied in high resolution imaging both ex vivo and in vivo samples.In system application, we have got a suit of flow in samples preparation both ex vivo and in vivo. To get low scattering, transparent ex vivo samples such as mice heart, colon and kidney, we processed the samples by clean, fixture, dehydrate and clear. To get in vivo samples such as drosophila melanogaster pupae, we processed the samples by clean and fixture. We could get ex vivo and in vivo OPT samples easy by this flow. During the experiments processing, we found the depth of field (DOF) in OPT is limited by its high magnification microscope, so the imaging quality would become poorly when the scale of samples greater than DOF. We acquire a series of projections at the same angle but different focal plane by moving the sample along the axis of EMCCD. Then, we obtain an optimal image by mean square error function at each angle. At last, we reconstruct the3D image by optimal images.