Acquisition of a Laser Scanning Confocal Microscope for Natural Science Research and Teaching

中文摘要美国国家科学基金会已授予史蒂文·W. Runge和同事在中央阿肯色州大学（UCA）获得激光扫描共聚焦显微镜（LSCM）。 该仪器能够捕获用荧光标记物标记的细胞和细胞结构的图像，其分辨率是其他类型的光学显微镜无法比拟的。 LSCM从各种活组织和准备好的组织中获取光学切片和构建3D图像以及对活细胞和组织进行时程分析的能力将影响加州大学的广泛研究项目。 将采用LSCM的研究范围从复杂组织的3D重建，如活跃生长的植物芽和大鼠和海洋蜗牛的大脑，到跟踪培养细胞和天然组织内部结构的运动。 总的来说，这种仪器的收购将提升教师和学生的研究在UCA的复杂性的新水平。 这将使研究人员能够解决以前无法解决的问题。 在UCA而不是在遥远的机构进行这项工作的一个重大好处是，我们的本科生参与研究的机会增加。 我们的研究生（硕士水平）也将获得重要的经验和技能，这将促进未来的研究和专业学习。LSCM将用于解决各种各样的自然科学研究问题。 其中包括：参与对紫外光反应的植物色素的快速测定和定位;哺乳动物脑白色物质中神经元的形态学表征;跟踪钾通道胞外插入肠膜用于海洋腹足类动物的呼吸调节;海洋腹足类动物整个脑中参与行为决定的突触连接的可视化;理解机制，在大鼠小脑成熟过程中浦肯野细胞的选择性死亡的基础;跟踪在开花植物中指导花序结构的发育信号的运动;检查植物中的分泌管和挖沟昆虫对胶乳/油分布的影响;以及比率成像以监测细胞内pH和在经历细胞凋亡的培养细胞中Na+/H+交换剂的囊泡运输的时程成像。 所采用的方法包括观察植物组织中的自发荧光分子和结构，用荧光抗体标记结构，用荧光分子填充单个脑细胞以识别与其他细胞的连接，以及用荧光标记物标记细胞表面蛋白以跟踪这些蛋白质在不同条件下进出细胞表面的运动。LSCM将是UCA科学教学的一个有价值的补充。 共聚焦显微镜将帮助学生可视化生物学中复杂的3D过程，并对活细胞进行测量，这将是教学的宝贵资产。 例如，青蛙胚胎发育的各个阶段将被记录为一组3D的时间过程图像，以跟踪这一动态过程。 然后，这些图像将在课堂上进行分析。 LSCM制作的令人惊叹的图像也将被用来通过各种推广计划和以UCA校园为中心的阿肯色州教育媒体来激发高中学生和教师对科学的兴趣。 该大学约有8，500名学生，其中62%为女性，19%为少数民族，后者的比例在过去十年中稳步上升。 收购LSCM将加强教师的研究能力，支持强大的本科研究项目，吸引更多的女性和少数民族学生从事科学事业，提高公众对科学的欣赏，并支持历史上支持不足的地理区域的项目。

AbstractA grant has been awarded to Dr. Steven W. Runge and colleagues at the University of Central Arkansas (UCA) to obtain a Laser Scanning Confocal Microscope (LSCM). This instrument has the ability to capture images of cells and cellular structures that are tagged with fluorescent markers with a resolution unmatched by other types of light microscopy. The ability of the LSCM to acquire optical sections and construct 3-D images from a variety of live and prepared tissues and to perform time-course analyses on living cells and tissues will impact a wide range of research programs at UCA. Investigations that will employ the LSCM range from 3-D reconstruction of complex tissues such as actively growing plant shoots and brains of rats and marine snails to tracking the movement of structures inside of cultured cells and native tissues. Collectively, acquisition of this instrument will elevate faculty and student research at UCA to new levels of sophistication. This will allow researchers to address questions that previously were inaccessible. A significant benefit of having this work conducted at UCA, rather than at distant institutions, will be the heightened opportunities for our undergraduates to become involved in research. Our graduate students (master's level) will also acquire important experience and skills that will promote future research and professional study.The LSCM will be used to address a wide variety of research questions in the natural sciences. These include: the rapid assay and localization of plant pigments involved in responses to UV-light; morphological characterization of neurons in the white matter of mammalian brains; tracking the exocytic insertion of potassium channels into intestinal membranes for osmoregulation in marine gastropods; visualization of synaptic connections involved in behavioral decisions in whole brains of marine gastropods; understanding mechanisms that underlie the selective death of Purkinje cells during maturation of the rat cerebellum; tracking the movement of developmental signals that direct inflorescence architecture in flowering plants; examination of secretory canals in plants and the effects of trenching insects on latex/oil distribution; and ratio imaging to monitor intracellular pH and time-course imaging of vesicular trafficking of Na+/H+ exchangers in cultured cells undergoing apoptosis. Methodologies to be employed include visualization of autofluorescent molecules and structures in plant tissues, tagging stuctures with fluorescent antibodies, filling individual brain cells with fluorescent molecules to identify connections to other cells, and labeling cell-surface proteins with fluorescent markers to follow the movement of these proteins to and from the cell surface under different conditions.In addition to research, the LSCM will be a valuable addition to science instruction at UCA. Confocal microscopy will help students visualize complex 3-D processes in biology and make measurements on living cells that will be invaluable assets to teaching. For instance, stages of frog embryo development will be recorded as a set of 3-D, time-course images that trace this dynamic process. These images will then be made available for analysis in the classroom. The stunning images produced by LSCM will also be used to generate interest in the sciences among high school students and teachers through various outreach programs and the Arkansas education media centered here on the UCA campus. The University's enrollment of about 8,500 students is 62% female and 19% minority, with the later group steadily increasing in proportion over the last decade. Acquisition of the LSCM will strengthen faculty research capabilities, support strong undergraduate research programs, attract more women and minority students to careers in the sciences, increase public appreciation of science, and support programs in a historically under-supported geographic area.