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.

这项重大研究仪器拨款支持购买荧光立体显微镜和成像系统，以加强科罗拉多学院生物系 Darrell J. Killian 博士和 Nancy N.Huang 博士的研究项目。活体模型生物体中荧光分子的实时可视化已成为一项不可或缺的技术，因为它可以直接观察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性的变化如何决定性别特异性发育。 Killian 实验室致力于鉴定和表征影响流行模式生物秀丽隐杆线虫（C. elegans）（一种土生蛔虫）性别特异性发育的基因。秀丽隐杆线虫种群包含雄性和雌雄同体，它们具有一些明显的形态差异。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成为识别性别特异性发育缺陷的强大而敏感的工具。已分离出的突变体是形态正常的雄性或雌雄同体，但具有通常只在异性中发现的神经元。受这些突变影响的基因的鉴定及其随后的表征将为性别特异性发育的遗传基础提供新的见解。荧光立体显微镜和成像对于使用性别特异性细胞类型的荧光标记在体内检测到的性缺陷的观察和记录至关重要。黄实验室对如何建立细胞身份或细胞命运规范感兴趣。MEX-3 是一种蛋白质，在线虫胚胎发育过程中在细胞命运规范中发挥重要作用。 MEX-3 蛋白最初存在于胚胎的所有细胞中，但在四细胞阶段从后细胞中逐渐耗尽。 MEX-3 分布的这种变化是胚胎前/后轴建立所必需的，因此了解 MEX-3 在胚胎中的定位非常重要。初步证据表明，MEX-3 的主动降解方式取决于两种不同类型的蛋白质修饰（称为磷酸化和泛素化）。MEX-3 蛋白质内这些修饰的特定位点的鉴定以及调节这些修饰的基因的鉴定都很感兴趣。当 MEX-3 中的特定位点突变或线虫基因组中的其他基因被破坏时，荧光显微镜将能够对荧光标记的 MEX-3 的稳定性进行体内研究。 科罗拉多学院是洛基地区唯一一所全国排名的文理学院美国山区，招收来自美国各地和国际的约 2000 名本科生。生物系坚定地致力于与本科生合作开展原创性研究，荧光显微镜和成像系统每年将为在 Killian 和 Huang 实验室工作的约 6-10 名本科生提供重要的研究和培训机会。此外，生物学课程中的许多课程都以基于探究的研究实验室经验为特色。具有此类经验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进与多门课程相关的实验室活动、演示和实验，每年将影响大约 200 名本科生。该成像系统将是一种特别有价值的教学工具，因为它使教师和一组学生能够同时在计算机显示器上观察同一样本。生物系还通过提供非专业课程在学院中发挥着强大的服务作用，这些课程培养了学生对基础生物学研究的强烈认识，并使学生能够就干细胞研究、基因治疗、转基因等许多生物学热门话题做出明智的决定和意见。生物和环境问题。荧光体视显微镜将为非专业学生提供独特而令人兴奋的机会，让他们亲身观察前沿科学。最后，科罗拉多学院积极寻求增强校园的多样性和最先进仪器的可用性，例如荧光体视显微镜和成像系统，将有助于吸引代表性不足的少数族裔和女性参与生物学研究。