Optical projection tomography scanner for the three-dimensional analysis of anatomical and gene expression data

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An increasing number of research groups in the department are investigating fundamental developmental and physiological processes in relatively large and thick samples (plant organs, vertebrate embryos), using markers of genetic expression (e.g. fluorescently tagged genes and proteins). Given the shape of the samples, gene expression data need to be acquired in three dimensions (3D). Current microscopy facilities in the department may be used to obtain three dimensional data in very small samples (e.g. confocal microscopy). However, acquiring quantitative 3D data from larger, thicker samples (from a millimeter to over a centimeter in diameter) involves serial sectioning and is therefore destructive, prone to alignment errors, potentially damaging to the samples, and very time consuming. The research of the applicants is therefore significantly delayed. The applicants propose to expand the departmental microscopy facility through the purchase of an Optical Projection Tomography (OPT) scanner. This new device recently developped in the UK is still very rare in laboratories at the international level. OPT can be used for the fast acquisition of 3D expression and anatomical data from whole samples up to 15mm in diameter with cellular detail. Semi transparent samples such as fish embryos can also be observed live. The OPT will allow the applicants to 1) Study the interactions between gene expression and morphology, which remains one of the grey areas of developmental biology, 2) Investigate the interactions between developmental and physiological processes such as respiration, 3) Investigate the effect of environmental toxicants on gene expression during development, and 4) Investigate gene regulation in the brain. Analyses of volumetric data using OPT will provide exciting new insights into physiological and developmental processes and will give the institution a leading edge in the 3D analysis of biological processes.