Acquisition of Laser Scanning Confocal Microscope

获取激光扫描共焦显微镜

• 批准号: 9413734
• 负责人: Melody V. S. Siegler
• 金额: $ 10.43万
• 依托单位: Emory University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-09-01 至 1996-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9413734&HistoricalAwards=false
• 关键词: Acquisition; Laser; Scanning; Confocal; Microscope

Funds are requested by the Emory University Biology Department for the purchase of a confocal microscope. The purchase will include a BioRad MRC-1000 confocal laser scanning imaging system and a Nikon Optiphot-2 microscope component. This system is unsurpassed in terms of specimen brightness, image resolution and ease of analysis. These capabilities are required by Biology Department labs that need to perform state of the art analyses of protein distribution in complex tissues. Principally four NSF-funded labs will use the equipment for research and training activities, although it is anticipated that wider departmental use will occur periodically. Three of the labs are investigating various aspects of insect neurogenesis (Ellis, Siegler and Yedvobnick) while the fourth lab is investigating sperm morphogenesis and migration in C. elegans (L'Hernault). The Ellis lab will utilize confocal microscopy to characterize the spatial distribution of the extramacrochaetae (emc) protein during embryonic development. Originally characterized for its role in achaete-scute regulation during sensory organ development, emc is now known to function more widely during development. Preliminary studies suggest that emc is expressed throughout the embryonic germ layers. The precise relationship between emc expression and achaete-scute expression will be characterized in the embryo. The Siegler lab will use confocal microscopy in conjunction with immunohistochemistry to identify neurons of known birthdate according to type, and thus test hypotheses about birth order within identified CNS lineages of the grasshopper. The Yedvobnick lab studies lateral inhibition and associated signal transduction to the nucleus during embryonic neurogenesis in Drosophila. They will utilize the equipment to investigate cytoplasmic-nuclear partitioning of the product of the neurogenic locus mastermind. They will also investigate potential physical interactions between cytoplasmic mastermind protein and the transmembranous Notch protein via colocalization assays. The L'Hernualt lab studies sperm morphogenesis that initiates as a multicellular syncitium. Confocal immunofluorescence will allow use of available antibodies to localize Golgi-derived membrane proteins within this complex three dimensional array. In addition, wild type and mutant sperm motility within the hermaphrodite reproductive tract will be examined by confocal microscopy, using already developed fluorescent tagging techniques. It is clear that conventional fluorescent microscopic techniques do not provide sufficient spatial detail to study these C. elegans sperm phenomena. The equipment will also be used extensively for the training of undergraduate and graduate students. The Biology Department has a sophisticated program for involving undergraduates in research. Beyond independent research Emory undergraduates are offered advanced project labs in Genetics, Neurobiology and Physiology; Development is planned for next year. In addition to academic year projects, summer research programs funded from NSF REU and the Howard Hughes Medical Institute bring in students from Emory and many other institutions. Although undergraduate use of the equipment will mostly be restricted to research purposes we anticipate incorporation of confocal microscopy demonstrations into the advanced project labs. Graduate students from programs including Neuroscience, Cell and Developmental Biology, and Genetics and Molecular Biology often rotate through Biology Department labs and several have joined the NSF-funded labs described above. Their utilization of the equipment should greatly enhance the quality of research output from the Biology Department.

埃默里大学生物系为购买共焦显微镜申请资金。此次收购将包括一个BioRad MRC-1000共聚焦激光扫描成像系统和一个尼康Optiphot-2显微镜组件。该系统在样品亮度、图像分辨率和分析简易性方面都是无与伦比的。这些能力是生物系实验室需要的，它们需要执行复杂组织中蛋白质分布的最先进分析。主要由NSF资助的四个实验室将使用这些设备进行研究和培训活动，尽管预计部门将定期更广泛地使用这些设备。其中三个实验室正在研究昆虫神经发生的各个方面(埃利斯、西格勒和叶德沃布尼克)，而第四个实验室正在研究线虫(L的赫尔诺)的精子形态发生和迁移。埃利斯实验室将利用共聚焦显微镜来表征胚胎发育过程中大毛外(EMC)蛋白的空间分布。EMC最初的特点是在感觉器官发育过程中调节毛刺，现在已知在发育过程中发挥更广泛的作用。初步研究表明，EMC在整个胚胎胚层都有表达。在胚胎中，EMC的表达和无毛皮肤的表达之间的确切关系将被表征。西格勒实验室将使用共聚焦显微镜和免疫组织化学相结合的方法，根据类型识别已知出生日期的神经元，从而在已确定的蝗虫中枢神经系统谱系中测试有关出生顺序的假设。叶德沃布尼克实验室研究果蝇胚胎神经发生过程中的侧向抑制和相关的核信号转导。他们将利用该设备研究神经源性基因座策划者的产物的细胞质-核分配。他们还将通过共定位分析来研究细胞质主脑蛋白和跨膜Notch蛋白之间的潜在物理相互作用。L的Hernualt实验室研究精子的形态发生，这种形态起源于一个多细胞联合体。共聚焦免疫荧光将允许使用可用的抗体来定位高尔基体衍生的膜蛋白在这个复杂的三维阵列中。此外，将使用已经开发的荧光标记技术，通过共聚焦显微镜检查两性生殖道内的野生型和突变精子的运动能力。很明显，传统的荧光显微镜技术不能提供足够的空间细节来研究这些线虫精子现象。该设备还将广泛用于本科生和研究生的培训。生物系有一个让本科生参与研究的复杂计划。除了独立研究，埃默里大学还为本科生提供遗传学、神经生物学和生理学方面的高级项目实验室；开发计划在明年进行。除了学年项目外，由NSF REU和霍华德·休斯医学院资助的暑期研究项目还吸引了来自埃默里和许多其他机构的学生。虽然本科生对这些设备的使用将主要限于研究目的，但我们预计共焦显微镜演示将被纳入高级项目实验室。神经科学、细胞和发育生物学、遗传学和分子生物学等专业的研究生经常在生物系的实验室中轮换，有几个已经加入了上述由NSF资助的实验室。他们对设备的使用应该会大大提高生物系的研究成果的质量。