MRI: Acquisition of a Fluorescence Stereomicroscope to Enhance Undergraduate Research at Colorado College

MRI：购买荧光立体显微镜以加强科罗拉多学院的本科生研究

• 批准号: 1126054
• 负责人: Darrell Killian
• 金额: $ 5.95万
• 依托单位: Colorado College
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-01 至 2013-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1126054&HistoricalAwards=false
• 关键词: MRI; Acquisition; Fluorescence; Stereomicroscope; Enhance

This Major Research Instrumentation grant supports the acquisition of a fluorescence stereomicroscopeand an imaging system to enhance the research programs of Dr. Darrell J. Killian and Dr. Nancy N.Huang in the Biology Department at Colorado College. The real-time visualization of fluorescentmolecules in living model organisms has become an indispensable technique because it allows the directobservation and documentation of changes in gene activity, the localization of proteins within a cell, andthe tracking of cells and tissues during development. An important question in biology is how changes ingene activity determine sex-specific development. The Killian Lab is interested in the identification andcharacterization of genes that affect sex-specific development in the popular model organismCaenorhabditis elegans (C. elegans), a soil-dwelling roundworm. C. elegans populations contain malesand hermaphrodites, which have some obvious morphological differences. However, sex-specific celltype differences in the nervous system are not easy to see. Fluorescent markers for sex-specific celltypes have emerged as powerful and sensitive tools for the identification of defects in sex-specificdevelopment. Mutants have been isolated that are morphologically normal males or hermaphrodites, butwith neurons that are normally only found in the opposite sex. The identification of the genes affected bythese mutations and their subsequent characterization will provide new insights into the genetic basis ofsex-specific development. Fluorescence stereomicroscopy and imaging are essential for the observationand documentation of sexual defects that are detected in vivo using fluorescent markers for sex-specificcell types. The Huang Lab is interested in how cellular identity, or cell fate specification, is established.MEX-3 is a protein that plays an important role in cell fate specification during C. elegans embryonicdevelopment. The MEX-3 protein is initially present in all cells of the embryo but becomes depleted fromthe posterior cells by the four-cell stage. This change in MEX-3 distribution is required for theestablishment of the anterior/posterior axis of the embryo, thus it is important to understand how MEX-3 islocalized in the embryo. Preliminary evidence suggests that MEX-3 is actively degraded in a manner thatis dependent upon two different types of protein modifications termed phosphorylation and ubiquitination.The identification of the specific sites of these modifications within the MEX-3 protein and theidentification of genes that regulate these modifications are of great interest. Fluorescence microscopywill enable an in vivo study of the stability of fluorescently tagged MEX-3 when specific sites in MEX-3 aremutated or when other genes in the C. elegans genome are disrupted.Colorado College is the only nationally ranked liberal arts college in the Rocky Mountain region of theUnited States and enrolls approximately 2000 undergraduates from all over the United States andinternationally. The Biology Department has a strong commitment to conducting original research withundergraduate collaboration and the fluorescence microscope and imaging system will provide importantresearch and training opportunities for approximately 6-10 undergraduates working in the Killian andHuang labs each year. In addition, many courses in the Biology curriculum feature inquiry-basedresearch lab experiences. Students with such experiences are best equipped to handle any postgraduatechallenges that they might face. The equipment will augment and improve laboratory activities,demonstrations, and experiments associated with several courses and will impact approximately 200undergraduates per year. The imaging system will be a particularly valuable teaching tool because it willenable the instructor and a group of students to simultaneously observe the same sample on a computermonitor. The Biology Department also plays a strong service role in the college by offering non-majorscourses that foster a strong appreciation for basic biology research and empower students to makeinformed decisions and opinions on many popular topics in biology such as stem cell research, genetherapy, genetically modified organisms, and environmental issues. The fluorescence stereomicroscopewill provide non-majors with unique and exciting opportunities to observe cutting-edge science first-hand.Finally, Colorado College actively seeks to enhance diversity on campus and the availability of state-ofthe-art instrumentation, such as a fluorescence stereomicroscope and imaging system, will help attractunderrepresented minorities and women to participate in biology research.

这项主要研究仪器补助金支持荧光立体显微镜和成像系统的收购，以加强达雷尔J.基利安博士和南希N.黄博士在科罗拉多学院生物系的研究计划。荧光分子在活体模型生物体中的实时可视化已经成为一项不可或缺的技术，因为它可以直接显示和记录基因活性的变化、细胞内蛋白质的定位以及发育过程中细胞和组织的跟踪。生物学中的一个重要问题是基因活动的变化如何决定性别特异性发育。Killian实验室对流行模式生物秀丽隐杆线虫（C. habditis elegans）中影响性别特异性发育的基因的鉴定和表征感兴趣。elegans），一种土栖蛔虫。C.线虫种群中存在雄性和雌雄同体，形态上存在明显差异。然而，神经系统中性别特异性细胞类型的差异并不容易看到。性别特异性细胞类型的荧光标记已经成为鉴定性别特异性发育缺陷的强大而敏感的工具。已经分离出的突变体在形态上是正常的男性或雌雄同体，但其神经元通常只存在于异性体内。这些突变影响的基因的鉴定及其随后的特征将为性别特异性发育的遗传基础提供新的见解。荧光体视显微镜和成像对于观察和记录性缺陷是必不可少的，这些性缺陷是使用性别特异性细胞类型的荧光标记物在体内检测的。Huang实验室对细胞身份或细胞命运规范是如何建立的感兴趣。MEX-3是一种在C.线虫胚胎发育MEX-3蛋白最初存在于胚胎的所有细胞中，但到了四细胞阶段，后部细胞中的MEX-3蛋白逐渐减少。MEX-3分布的这种变化是建立胚胎前/后轴所必需的，因此了解MEX-3在胚胎中的定位是很重要的。初步证据表明，MEX-3的降解依赖于磷酸化和泛素化两种不同类型的蛋白质修饰，对MEX-3蛋白中这些修饰的特异位点的鉴定以及对调控这些修饰的基因的鉴定具有重要意义。当MEX-3中的特定位点发生突变或C.科罗拉多学院是美国落基山脉地区唯一一所全国排名的文科学院，招收了来自美国各地和国际上的大约2000名本科生。生物系有一个坚定的承诺，进行原创性研究与本科生的合作和荧光显微镜和成像系统将提供重要的研究和培训机会，每年约6-10名本科生在基利安和黄实验室工作。此外，生物学课程中的许多课程都有基于探究的研究实验室体验。有这样经历的学生最有能力应对他们可能面临的任何研究生挑战。这些设备将增强和改善与几门课程相关的实验室活动、演示和实验，每年将影响大约2000名本科生。该成像系统将是一个特别有价值的教学工具，因为它将使教师和一组学生同时观察同一个样品在计算机显示器上。生物学系还通过提供非专业课程，培养对基础生物学研究的强烈赞赏，并使学生能够对生物学中的许多热门话题（如干细胞研究，基因治疗，转基因生物和环境问题）做出明智的决定和意见，在学院中发挥着强大的服务作用。最后，科罗拉多学院积极寻求提高校园的多样性和最先进的仪器，如荧光立体显微镜和成像系统的可用性，将有助于吸引未被充分代表的少数民族和妇女参与生物学研究。