Kumar Sunil, Lockwood Nicola, Ramel Marie-Christine, Correia Teresa, Ellis Matthew, Alexandrov Yuriy, Andrews Natalie, Patel Rachel, Bugeon Laurence, Dallman Margaret J, Brandner Sebastian, Arridge Simon, Katan Matilda, McGinty James, Frankel Paul, French Paul M W
Department of Physics, Imperial College London, London SW7 2AZ, UK.
Division of Medicine, University College London, London WC1E 6JF, UK.
Oncotarget. 2016 Jul 12;7(28):43939-43948. doi: 10.18632/oncotarget.9756.
We describe a novel approach to study tumour progression and vasculature development in vivo via global 3-D fluorescence imaging of live non-pigmented adult zebrafish utilising angularly multiplexed optical projection tomography with compressive sensing (CS-OPT). This "mesoscopic" imaging method bridges a gap between established ~μm resolution 3-D fluorescence microscopy techniques and ~mm-resolved whole body planar imaging and diffuse tomography. Implementing angular multiplexing with CS-OPT, we demonstrate the in vivo global imaging of an inducible fluorescently labelled genetic model of liver cancer in adult non-pigmented zebrafish that also present fluorescently labelled vasculature. In this disease model, addition of a chemical inducer (doxycycline) drives expression of eGFP tagged oncogenic K-RASV12 in the liver of immune competent animals. We show that our novel in vivo global imaging methodology enables non-invasive quantitative imaging of the development of tumour and vasculature throughout the progression of the disease, which we have validated against established methods of pathology including immunohistochemistry. We have also demonstrated its potential for longitudinal imaging through a study of vascular development in the same zebrafish from early embryo to adulthood. We believe that this instrument, together with its associated analysis and data management tools, constitute a new platform for in vivo cancer studies and drug discovery in zebrafish disease models.
我们描述了一种通过利用具有压缩传感的角度复用光学投影断层扫描（CS-OPT）对活体非色素成年斑马鱼进行整体三维荧光成像来研究体内肿瘤进展和血管系统发育的新方法。这种“介观”成像方法弥合了既定的约微米分辨率的三维荧光显微镜技术与约毫米分辨率的全身平面成像和扩散断层扫描之间的差距。通过CS-OPT实施角度复用，我们展示了在成年非色素斑马鱼中对一种可诱导的荧光标记肝癌基因模型进行体内整体成像，该模型还呈现荧光标记的血管系统。在这个疾病模型中，添加化学诱导剂（强力霉素）可驱动免疫健全动物肝脏中eGFP标记的致癌K-RASV12的表达。我们表明，我们新颖的体内整体成像方法能够在疾病进展过程中对肿瘤和血管系统的发育进行非侵入性定量成像，我们已通过包括免疫组织化学在内的既定病理学方法对其进行了验证。我们还通过对同一斑马鱼从早期胚胎到成年期的血管发育研究证明了其纵向成像的潜力。我们相信，该仪器及其相关的分析和数据管理工具构成了一个用于斑马鱼疾病模型体内癌症研究和药物发现的新平台。
