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

这个提案的目标是在这个詹姆斯A。佛罗里达州坦帕的海利退伍军人医院。 BAUDIITY是一种革命性的新系统，可以看到死后组织内部。到目前为止，研究人员 不得不将组织切成薄片，然后观察组织的小区域。与 完整的组织，如整个大脑，脊髓，心脏或其他组织， 透明，使研究人员能够研究亚细胞结构，神经网络，特定的 蛋白质、生物分子复合物和基因表达谱分析在单个细胞的分辨率。为 这是第一次，研究人员可以看到整个大脑，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 疾病和中风。MRI和其他成像方法对活体组织有好处，但它们不能提供 由于在单细胞水平上的分辨率，他们不能识别轻度创伤性脑损伤，也不能识别 这些变化都是在疾病全面发展之前发生的。MRI不适用于小动物，如大鼠和 老鼠，神经科学研究的首选动物。作为评审员，我们现在看到赠款 其他机构在其项目中使用的可扩展性。事实上赠款使用传统解剖学和 神经解剖学的方法正在被拒绝。为了保持我们的竞争优势， 清楚起见该设备将实现以下具体目标：i）图像神经回路， 由光遗传学刺激激活的分子途径和机制，ii）鉴定这些分子途径和机制中的缺陷， 对酒精和药物成瘾以及特定疾病作出反应的回路和途径，iii）确定 这些通路在对心血管疾病、肾脏疾病、创伤性 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.