MRI: Acquisition of New Fluorescence/DIC Microscope with Computing Station for Research and Student Training

中央华盛顿大学(CWU)是一所文凭和硕士授予机构，位于西雅图以东约110英里的农村农业区，在喀斯喀特山脉的另一边。华威大学拥有约9000名学生，其中约20%的学生是少数民族。来自生物和化学系的五名研究人员要求购买仪器，以增强他们现有的显微镜设施。这项收购将极大地加强华威大学的教学、培训和研究基础设施。该专用仪器是一种新型徕卡DM5500复合显微镜系统，具有荧光和差分干涉对比(DIC)功能。该方案包括一个隔振台和一个计算工作站，这是充分利用显微镜能力所必需的。为了促进多种颜色的荧光可视化，显微镜将配备与荧光兼容的物镜；一个广谱(通过红外的紫外线)高强度光源；以及多个滤光片，允许特定波长用于不同的实验。几乎所有的研究人员都将使用荧光显微镜来分析荧光标记的细胞器，如细胞核、线粒体和轴突。它还将用于表征存在于有机体细胞中的多种基因和蛋白质，从极端细菌到微小的蠕虫，再到发育中的雏鼠和成年小鼠的大脑。DIC功能将允许研究人员可视化(并最终测量)组织内或细胞内未经荧光或组织学染色处理的结构的大小。这种光学能力对于研究各种真菌生长的研究人员以及那些需要可视化蠕虫和雏鸡大脑中未染色的细胞和组织的人来说是必要的。与荧光功能类似，这些DIC光学设备需要在显微镜的光路中使用高度专业化的物镜、滤光片和棱镜。为了记录实验结果，数码相机和一台计算机是必要的。由于一些调查人员将使用明场(普通白色)光拍摄，而另一些调查人员将拍摄荧光图像，因此需要两种不同的相机。每一种都是专门用于高图像质量和准确、定量地表示亮场或荧光样本。此外，对获得的图像执行的许多分析都需要强大的计算能力。所要求的计算机有存储空间来存储将要收集的许多大图像，它将配备最先进的软件来执行以下功能：去卷积是一种分析样本的多个焦面的技术，以数字减去未聚焦的材料，以高度澄清最终图像。荧光强度的定量将允许自动计数和测量标记结构的大小，如细菌、轴突和线粒体；它还将允许比较试验组之间的生化和DNA差异。显微镜舞台的计算机控制允许对舞台运动进行校准和量化，以测量不能在一张图像中完全可视化的三维样本的大小。参与这项提议的研究人员正在研究各种生物学领域，如细胞和发育神经科学、真菌和真菌样原生生物的系统学和生物学、极端环境的微生物生态学以及线粒体保护免受氧化应激的机制。这些领域中的每一个都要求科学家能够将分子和生化变化与细胞结构和行为的变化联系起来。需要最先进的显微镜仪器来分析和记录细胞和亚细胞过程，如发育和行为适应过程中神经元连接的变化，已知和新表征的真菌和原生生物之间的形态差异，病毒与极端微生物的相互作用，以及线粒体对压力的反应变化。获得这一仪器将使调查人员能够在这些领域建立和扩大他们的研究，并继续在他们的实验室培训本科生和研究生。事实上，华威大学的生物系和化学系能够将许多本科生纳入正在进行的研究中。本科生发现，与至少一名研究教授接触相对容易，通常通过演讲和出版来传播这项研究。此外，教师致力于在实验室课程中融入探究式教学和对学生的技术培训。购买这种仪器将使更多的本科生和硕士研究生在接受培训成为未来的科学家时，能够接触到最先进的研究显微镜。华威大学建立并支持了几个项目，如本科生研究的科学荣誉计划以及研究和旅行的竞争性基金。华威大学也有外部资助的项目，如美国国家科学基金会资助的科学人才扩展计划和教育部的麦克奈尔学者计划，这两个计划都包括努力增加对科学界代表性不足群体的招聘和留住。CWU正在通过NSF支持的GK12 Grant，Yakima Waters在当地初中和高中实施跨学科分水岭研究，该项目将研究生研究员安排到学校，将探究性教育纳入课程。这项提案的调查人员积极参与了这些项目。

Central Washington University (CWU) is a Baccalaureate and Masters Granting Institution located in a rural agricultural region about 110 miles east of Seattle on the other side of the Cascade Mountains. CWU has a student population of about 9,000 students with about 20% minority enrollment. The five investigators from the departments of Biology and Chemistry are requesting acquisition of instrumentation to enhance their existing microscopy facility. This acquisition will greatly enhance the teaching, training and research infrastructure at CWU. The specific instrument is a new Leica DM5500 compound microscope system with fluorescence and differential interference contrast (DIC) capabilities. The proposal includes a vibration isolation table and a computational workstation that are necessary for utilizing the full capabilities of the microscope. To facilitate fluorescence visualization in multiple colors the microscope will be equipped with fluorescence-compatible objective lenses; a broad-spectrum (ultraviolet through infrared), high intensity light source; and multiple filters that allow specific wavelengths to be used for different experiments. Fluorescence microscopy will be used by nearly all the investigators to analyze fluorescently labeled cell organelles such as nuclei, mitochondria and axons. It will also be used to characterize multiple genes and proteins that are present in the cells of organisms ranging from extremophile bacteria to microscopic worms to developing chick and adult mouse brains. The DIC capability will allow investigators to visualize (and ultimately measure sizes of) structures within a tissue or within cells that have not been treated with a fluorescent or histological stain. This optical capability is necessary for investigators who study the growth of diverse fungi and those who need to visualize unstained cells and tissue in the worm and chick brain. Similar to the fluorescence capabilities, these DIC optics require highly specialized objective lenses, filters and prisms in the light path of the microscope. In order to document results of experiments, digital cameras and a computer are necessary. Because some investigators will photograph with brightfield (normal white) light and others will photograph fluorescent images, two different cameras are required. Each is specialized for high image quality and accurate, quantitative representation of the brightfield or fluorescent specimen. Additionally, intense computational power is required for many of the analyses that will be performed on the images obtained. The requested computer has storage space for the many large images that will be collected, and it will be equipped with state-of-the-art software for performing the following functions: Deconvolution is a technique to analyze multiple focal planes of a specimen to digitally subtract unfocussed material in order to highly clarify the final image. Quantitation of fluorescence intensity will allow automated counting and measurement of sizes of labeled structures such as bacteria, axons, and mitochondria; it will also allow comparison of biochemical and DNA differences among experimental groups. Computerized control of the microscope stage allows stage movements to be calibrated and quantitated in order to measure the size of a three-dimensional specimen that cannot be visualized completely in one image.The investigators involved in this proposal are studying areas of Biology that are as diverse as cell and developmental neuroscience, systematics and biology of fungi and fungus-like protists, microbial ecology of extreme environments and mitochondrial mechanisms of protection from oxidative stress. Each of these areas requires the scientists to be able to correlate molecular and biochemical changes with changes in cell structure and behavior. The most advanced microscopy instrumentation is required to analyze and document cellular and subcellular processes, such as changes in neuronal connections during development and behavioral adaptation, morphological differences among known and newly characterized fungi and protists, viral interactions with extreme microbes, and alterations to mitochondria in response to stress. Acquisition of this instrument will enable the investigators to establish and extend their research in these areas and to continue to train undergraduate and graduate students in their laboratories. Indeed, the Biology and Chemistry departments at CWU are able to incorporate many undergraduate students in the ongoing research. Undergraduates find it relatively easy to get involved with at least one research professor and typically disseminate this research through presentations and publication. Furthermore, the faculty are committed to incorporating inquiry-based teaching and technical training of students in laboratory classes. The acquisition of this instrumentation will allow even more undergraduates and masters-level graduate students to have access to state-of-the-art research microscopes as they train to become future scientists. CWU has established and provided support for several programs such as the Science Honors Program for undergraduate research and competitive funds for research and travel. CWU also has externally funded programs such as the NSF-funded Science Talent Expansion Program and Department of Education's McNair Scholars Program, both of which include efforts to increase recruitment and retention of underrepresented groups in science. CWU is implementing interdisciplinary watershed research into local middle and high schools through the NSF-supported GK12 Grant, Yakima WATERS, which places graduate fellows into schools to incorporate inquiry-based education into the curriculum. The investigators on this proposal are actively involved in these programs.