Whole body tomography of fluorescent proteins in vivo

描述(申请人提供)：荧光蛋白(FP)已成为阐明蛋白质在细胞内的功能、免疫和干细胞的生物分布以及评价体内候选药物的重要报告分子。整个动物体内荧光蛋白的体积检测和准确定量将极大地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促进的，这些技术损害了定量和任何体积成像能力。因此，这项提议的总体目标是在整个动物中开发全身定量荧光蛋白断层扫描(FPT)。这一发展需要1)使用新的扫描技术，2)发展适用于可见光和远红外线成像的光子传播理论模型，3)设计具有多光谱特性的计算高效的反转技术，4)询问能够产生高检测灵敏度的合适的荧光蛋白质。要实现高成像性能，尤为重要的是利用了FreesSpace 360度投影断层扫描原理和适当的自动荧光减影方案，与当前技术水平相比，它们带来了前所未有的成像性能。总体而言，我们假设，FPT赋予的成像准确性，结合荧光蛋白报告程序获得的高特异性和多功能性，以及新型红移结构提供的高检测灵敏度，可以彻底改变生物成像，并将FPT作为生物实验室体积全动物成像的首选方法。

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.