ShEEP Request for Keyence BZ-X800E All-in-One Automated Imaging System

本共享设备评估计划（ShEEP）提案的目的是要求Keyence BZX800E一体化自动成像系统将为研究人员提供一种能够： 厚组织切片的类共焦多叠层成像和实时聚焦;快速自动高 合成图像的放大图像采集和拼接; 以高分辨率成像的组织切片。控制软件简单直观。以最小的 培训一个新手用户可以扫描一系列组织切片从整个器官， 每个组织切片的合成物，并将所有合成物合并成单个三维渲染的组织切片。 整个组织巴尔的摩退伍军人医疗中心（BVAMC）提供的传统显微镜不具备 这些能力。 退伍军人事务部的项目包括一系列专业，如下所述。每个人都将受益于Keyence BZX800E 一体化自动成像系统。BZX800E显微镜优化了成像过程， 设计一个全自动的，易于使用的系统，捕捉高分辨率的图像，并允许研究人员 以更快地达到他们想要的结果。这些应用程序对于VA生物医学实验室非常有用 研究与开发服务（BLR & D）、临床科学研究与开发（CSR & D）和 康复研究与发展（RR & D）资助的研究人员。Tibor Kristian博士将作为 本提案的项目总监（PD）/主要研究者（PI）。克里斯蒂安博士的研究重点是 理解导致线粒体和最终细胞生物能量衰竭的损伤机制， 中风患者这种成像系统对于细胞内细胞器的成像和定量是必不可少的 中风引起的缺血性脑损伤后的损害。Bingren Hu，医学博士，博士正在验证一个新的假设 脑缺血-再灌注导致一系列导致失活的事件; 脑缺血-再灌注（IR）损伤的机制。许多抗体将用于目标1和2 胡博士的研究; BZX800显微镜是必要的成像和定量细胞内 IR脑损伤后细胞器损伤。Bogdan Stoica，医学博士将使用这个新系统来调查 创伤性脑损伤（TBI）。斯托伊卡博士的项目需要大量的慢性神经炎症成像 标记和二次损伤标记。塔帕斯·马卡尔博士的项目研究了7，8- 二羟基黄酮（7，8-DHF）;其具有有效的脑源性神经营养因子（BDNF）激动剂活性， 将在他的具体目标2和3中使用成像系统。Makar医生将决定DHF治疗是否 调节脑脊髓炎（EAE）小鼠模型脑和脊髓中的炎症和免疫 多发性硬化症的治疗方法。列奥纳多托内利博士的项目研究的压力 部署和暴露于创伤事件，使士兵面临比一般公众更大的风险， 出现心理障碍Tonelli博士的分析将包括邻近组织中的免疫组织化学。 部分：归巢标志物，趋化因子受体，小胶质细胞和巨噬细胞活化和/或 差异化-所有这些都可以通过Keyence BZ-X800查看。谢国锋，医学博士研究冒号 他的长期目标是通过靶向关键信号来开发创新，有效和安全的疗法。 结肠癌进展的转导分子。显微镜将允许谢博士检查 使用免疫荧光抗MMP7抗体的基质金属蛋白酶7（MMP7）表达模式 人结肠癌细胞和手术标本。受资助的项目构成了翻译研究的基础 这将改善退伍军人的治疗-可能产生新的治疗方法。

The objective of this Shared Equipment Evaluation Program (ShEEP) proposal is to request Keyence BZX800E All-in-One Automated Imaging System which will offer researchers an instrument that is capable of: confocal-like multi-stack imaging and real-time focusing of thick tissue sections; rapid automatic high- magnification image acquisition and stitching of composite images; and three-dimensional profiling of serial tissue sections imaged at high resolution. The control software is simple and intuitive. Thus, with minimal training a novice user can scan a series of tissue sections from an entire organ, generate high-resolution composites of each tissue section, and merge all composites into a single three-dimensional rendering of the entire tissue. Traditional microscopes available at the Baltimore VA Medical Center (BVAMC) do not possess these capabilities. The VA projects cover an array of specialties as described below. Each will benefit from Keyence BZX800E All-in- One Automated Imaging System. The BZX800E Microscope optimizes the imaging process by designing a fully automated, easy-to-use system that captures high-resolution images and allows researchers to achieve their desired results faster. These applications are of great use for VA Biomedical Laboratory Research and Development Service (BLR&D), Clinical Science Research and Development (CSR&D), and Rehabilitation Research and Development (RR&D) funded investigators. Tibor Kristian, Ph.D. will serve as the Project Director (PD)/ Principal Investigator (PI) for this proposal. Dr. Kristian’s study focuses on understanding injury mechanisms that lead to mitochondrial and ultimately cellular bioenergetic failure in stroke victims. This imaging system is essential for imaging and quantification of intracellular organelles damage following stroke- induced ischemic brain injury. Bingren Hu, M.D., Ph.D. is testing a novel hypothesis that brain ischemia- reperfusion leads to a cascade of events resulting in inactivation; as well as study novel mechanisms underlying brain ischemia-reperfusion (IR) injury. Many antibodies will be used in Aims 1 and 2 of Dr. Hu’s studies; and the BZX800 microscope is necessary for imaging and quantification of intracellular organelles damage following IR brain injury. Bogdan Stoica, M.D. will use this new system to investigate traumatic brain injury (TBI). Dr. Stoica’s project requires significant imaging of chronic neuroinflammation markers and secondary damaging markers. Tapas Makar, Ph.D.’s project studies the effects of 7, 8- dihydroxyflavone (7,8- DHF); which has potent brain derived neurotrophic factor (BDNF) agonist activity and will use the imaging system in his specific aims 2 and 3. Dr. Makar will determine whether DHF treatment modulates inflammation and immunity in the brain and spinal cord in encephalomyelitis (EAE), a murine model of Multiple sclerosis, using monoclonal antibodies. Leonardo Tonelli, Ph.D.’s project studies the stress of deployment and exposure to traumatic events that puts soldiers at a greater risk than the general public in developing psychological disorders. Dr. Tonelli’s analysis will consist of immunohistochemistry in adjacent sections for: homing markers, chemokine receptors, microglia and macrophage activation and/or differentiation- all of which can be viewed with the Keyence BZ-X800. Guofeng Xie, M.D. studies colon cancer and his long-term goal is to develop innovative, effective, and safe therapies by targeting pivotal signal transduction molecules underlying colon cancer progression. The microscope will allow Dr. Xie to examine the expression patterns of matrix metalloproteinase 7 (MMP7) using immunofluorescent anti-MMP7 antibody in human colon cancer cells and surgical specimens. The funded projects form the basis for translational studies that will improve treatment in Veterans- potentially yielding new therapeutics.