ShEEP Request for CLARITY Optimized Light sheet Microscope for high speed imaging of large clarified samples at high resolution

这项提案的目标是在佛罗里达州坦帕市的詹姆斯·A·黑利退伍军人医院安装Clarity。 Clarity是一种革命性的新系统，可以看到身体组织内部的情况。到目前为止，研究人员 必须将组织切成薄片，然后将组织的小区域可视化。使用 清晰度整个组织，如整个大脑、脊髓、心脏或其他组织， 透明的使研究人员能够研究亚细胞结构、神经元网络、特定的 蛋白质、生物分子复合体和单细胞分辨率的基因表达谱。为 第一次，研究人员可以可视化整个大脑、脊髓或其他组织的缺陷 与神经退行性疾病、创伤性脑损伤、衰老、癌症、心血管疾病有关 疾病和中风。核磁共振和其他成像方法对活组织有好处，但它们不能提供 在单细胞水平上的分辨率，他们不能识别轻微的创伤性脑损伤，也不能识别 在全面病理之前的变化。核磁共振不适用于小动物，如大鼠和 小鼠，神经科学研究中的首选动物。作为评审员，我们现在看到来自 其他在项目中使用清晰度的机构。事实上，使用传统解剖学和 神经解剖学方法被拒绝了。为了保持我们的竞争优势，必须安装 清晰度。该设备将完成以下具体目标：i)图像神经电路和 光遗传刺激激活的分子途径和机制，II)识别其中的缺陷 对酒精和药物成瘾和特定疾病的反应回路和通路，三)确定 这些通路因心血管疾病、肾脏疾病、创伤性疾病而被破坏 脑损伤、中风、脊髓损伤、阿尔茨海默病和帕金森病。

The goal of this proposal is to install CLARITY at this James A. Haley Veterans' Hospital in Tampa, FL. CLARITY is a revolutionary new system that can see inside post-mortem tissue. Until now researchers have had to slice the tissue into thin sections and then visualize small areas of the tissue. With CLARITY the entire tissue, such as a whole brain, spinal cord, heart or other tissue, is made transparent allowing researchers to investigate the subcellular structure, neuronal network, specific proteins, biomolecular complexes and gene expression profiling at the resolution of single cells. For the first time, researchers can visualize the entire brain, spinal cord, or other tissue for defects associated with neurodegenerative diseases, traumatic brain injury, aging, cancer, cardiovascular disease and stroke. MRI and other imaging methods are good for living tissue, but they do not provide resolution at the single cell level, they cannot identify mild traumatic brain injury, nor can they identify changes that precede full-blown pathology. MRI is not applicable to small animals, such as rats and mice, the animals of choice in neuroscience research. As reviewers, we are now seeing grants from other institutions that use CLARITY in their projects. Indeed, grants that use traditional anatomical and neuroanatomy methods are being rejected. To maintain our competitive edge it is essential to install CLARITY. This equipment will accomplish the following specific aims: i) image neural circuits and molecular pathways and mechanisms activated by optogenetic stimulation, ii) identify defects in these circuits and pathways in response to alcohol and drug addiction and specific diseases, iii) identify where these pathways are disrupted in response to cardiovascular disease, renal disease, traumatic brain injury, stroke, spinal cord injury, Alzheimer’s disease and Parkinson’s disease.