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

该部门越来越多的研究小组正在使用遗传表达的标记(例如荧光标记的基因和蛋白质)，在相对较大和较厚的样本(植物器官、脊椎动物胚胎)中调查基本的发育和生理过程。给定样本的形状，需要获取三维(3D)的基因表达数据。该部门现有的显微镜设备可用于获取非常小样本的三维数据(例如共焦显微镜)。然而，从更大、更厚的样品(直径从一毫米到一厘米以上)获取定量3D数据涉及连续切片，因此是破坏性的，容易出现对准错误，潜在地损害样品，并且非常耗时。因此，申请者的研究工作大大推迟了。申请者提议通过购买光学投影断层扫描仪来扩大部门的显微镜设施。这种最近在英国开发的新设备在国际水平的实验室中仍然非常罕见。OPT可用于快速获取直径达15 mm的完整样本的3D表达和解剖数据，以及细胞细节。半透明的样品，如鱼胚胎，也可以被活体观察到。OPT将允许申请者1)研究基因表达和形态之间的相互作用，这仍然是发育生物学的灰色区域之一；2)研究发育和生理过程(如呼吸)之间的相互作用；3)调查环境毒物对发育过程中基因表达的影响；以及4)研究大脑中的基因调控。使用OPT分析体积数据将为生理和发育过程提供令人兴奋的新见解，并将使该机构在生物过程的3D分析方面处于领先地位。

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.