Whole body tomography of fluorescent proteins in vivo

描述（由申请人提供）：荧光蛋白（FP）已成为阐明细胞内蛋白质功能、免疫细胞和干细胞生物分布以及体内评价候选药物的基本报告分子。在整个动物中荧光蛋白的体积检测和准确定量将大大提高我们监测体内生物过程的能力。然而，目前，全身荧光蛋白成像在很大程度上是由摄影技术，妥协的量化和任何体积成像能力。因此，本提案的总体目标是在整个动物中开发全身定量荧光蛋白质断层扫描（FPT）。这一发展需要1）使用新的扫描技术，2）光子传播的理论模型的进步，适用于在可见光和远红外成像，3）设计计算效率高的反演技术与多光谱特性和4）适当的荧光蛋白质，可以产生高检测灵敏度的询问。对于实现高成像性能特别重要的是利用自由空间360度投影断层摄影原理和适当的自动荧光减影方案，其导致与当前技术水平相比前所未有的成像性能。结合荧光蛋白报告子所实现的高特异性和多功能性以及新的红-移位的构建体可以彻底改变生物成像，并将FPT作为生物实验室中用于体积整体动物成像的选择方法进行推广。

DESCRIPTION (provided by applicant): Fluorescent Proteins (FP) have become essential reporter molecules for the elucidation of the function of proteins within cells, the bio-distribution of immune and stem cells and for evaluation of drug candidates in vivo. Volumetric detection and accurate quantification of Fluorescent Proteins in entire animals would greatly enhance our ability to monitor biological processes in vivo. Currently however, whole body fluorescent protein imaging is largely facilitated by photographic techniques that compromise the quantification and any volumetric imaging ability. The overall goal of this proposal is therefore to develop whole body, quantitative Fluorescence Protein Tomography (FPT) in entire animals. This development necessitates 1) the use of novel scanning technologies, 2) the advancement of theoretical models of photon propagation appropriate for imaging in the visible and far-red, 3) the design of computationally efficient inversion techniques with multispectral characteristics and 4) the interrogation of appropriate fluorescent proteins that can yield high detection sensitivity. Of particular importance for achieving high imaging performance is the utilization of freespace 360 degree projection tomographic principles and appropriate auto-fluorescent subtraction schemes that lead to unprecedented imaging performance compared to the current state of the art. Overall we hypothesize that the imaging accuracy imparted by FPT, combined with the high specificity and versatility achieved by fluorescent protein reporters and the high detection sensitivity afforded by novel red-shifted constructs can revolutionize biological imaging and propagate FPT as the method of choice in the biological laboratory for volumetric whole animal imaging.