Acquisition of Computer-Assisted Microscopy Imaging System for Cytogenetic Research and Training in Neuroscience and Reproductive Biology

获取计算机辅助显微成像系统，用于神经科学和生殖生物学的细胞遗传学研究和培训

• 批准号: 0116477
• 负责人: Diane Lee
• 金额: --
• 依托单位: California State University-Long Beach Foundation
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-01 至 2004-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0116477&HistoricalAwards=false
• 关键词: Acquisition; Computer; Assisted; Microscopy; Imaging

A grant has been awarded to Drs. Lee and Underwood at California StateUniversity Long Beach to acquire an imaging system comprised of a fluorescence-capable microscope with motorized stage, CCD camera, computer, and software. This system will provide unbiased stereology capabilities, as well as the ability to 1) visualize immunofluorescent labeled samples; 2) produce 3-dimensional (3-D)reconstructions; and 3) establish image databases accessible via the internet. The goal of Dr. Lee's research is to understand the effects of memoryformation and hormones on neurogenesis in the zebra finch hippocampus. Thehippocampus of adult birds and mammals retains the capacity for substantialplasticity including the birth of new cells (cytogenesis) and neurons(neurogenesis) in response to a number of environmental influences includinglearning and memory formation. Dr. Lee recently found that hippocampal cellsare also born as a result of injury and collect around the damaged site in thezebra finch brain. Injured brain areas become rich in aromatase as well, anenzyme that converts testosterone to estrogen. Estrogens, in turn, are knownto promote differentiation and survival of neurons. These intriguing resultsare being used to investigate factors influencing cytogenesis anddifferentiation of new cells into neurons and glia. The aim then is to exploitthe ability of the avian hippocampus to respond to environmental influences bytraining birds on learning tasks, lesioning the hippocampus, injecting themwith BrdU (a mitotic marker), and assessing the patterns of cell proliferation,migration, and/or differentiation through the use of immunohistochemicaltechniques. New BrdU-positive cells will be visualized and counted using theMicroBrightField StereoInvestigator imaging system; double labeling procedureswill be used to determine whether new cells are neurons or glia. The goal of Dr. Underwood's research is to understand the relationshipbetween chromosomal abnormalities, a biased sex ratio, and the evolution of"excess" males in the madrone butterfly. Dr. Underwood's research led to thediscovery of abnormalities in chromosome pairing during germ cell formation inthe madrone butterfly. One subspecies shows a normal complement of 26bivalents; however, another subspecies shows extreme variations from cell tocell within the same individual. The primary sex ratio of the abnormalsubspecies is 75% male while the normal subspecies appears to have a normal sexratio of 50% males. Butterflies have chromosomal sex determination.Spermatogenesis is extremely abnormal with multivalent chromosomal associationsand lagging chromosomes formed during metaphase and anaphase, and theproduction of micronuclei associated with sperm nuclei. NSF funding will also make it possible to digitize samples obtained from avariety of research projects. This will result in public databases to be usedby interested scientists with access to the internet and thus offers anunparalleled opportunity to share information both locally and globally.Individuals from multiple labs will perform procedures remotely thus broadeningthe scope of research and increasing the opportunity for publication anddissemination of results, while minimizing labor and the number of animals.Beyond the immediate research applications, instructors and their students willbe empowered through accessibility to data previously reserved for only aselect few. Image databases, accessed as classroom exercises, will exposestudents to topical and exciting new research as well as train them in the useof 21st century technology thus advancing science and giving them a competitiveedge.

加州州立大学长滩分校的Lee博士和Underwood博士获得了一笔赠款，用于购买一种成像系统，该系统由带有电动工作台的荧光显微镜、CCD相机、计算机和软件组成。该系统将提供无偏见的体视学能力，以及1)可视化免疫荧光标记的样本；2)产生3维(3-D)重建；3)建立可通过互联网访问的图像数据库。李博士的研究目标是了解记忆形成和激素对斑雀海马区神经发生的影响。成年鸟类和哺乳动物的海马体保留了物质可塑性的能力，包括新细胞(细胞发生)和神经元(神经发生)的诞生，以应对包括学习和记忆形成在内的许多环境影响。李博士最近发现，海马细胞也是因损伤而诞生的，并聚集在斑马雀大脑受损部位周围。受损的大脑区域也会富含芳香酶，芳香酶是一种将睾丸激素转化为雌激素的酶。雌激素反过来又被认为可以促进神经元的分化和存活。这些耐人寻味的结果正被用于研究影响细胞发生和新细胞分化为神经元和胶质细胞的因素。然后，目的是开发鸟类海马体对环境影响的反应能力，方法是在学习任务中训练鸟类，损毁海马体，给它们注射BrdU(有丝分裂标记物)，并使用免疫组织化学技术评估细胞增殖、迁移和/或分化的模式。新的BrdU阳性细胞将使用MicroBrightFieldStereoInvestigator成像系统进行可视化和计数；双重标记程序将用于确定新细胞是神经元还是胶质细胞。安德伍德博士的研究目标是了解染色体异常、性别比例偏向以及马德龙蝴蝶雄性数量“过多”的进化之间的关系。安德伍德博士的研究导致在马德龙蝴蝶生殖细胞形成过程中发现了染色体配对的异常。一个亚种表现出26个二价体的正常互补；然而，另一个亚种在同一个体内的不同细胞之间表现出极端的差异。异常亚种的主要性别比是75%的男性，而正常亚种的男性似乎有50%的正常性别比。蝴蝶有染色体性别决定，精子发生极度异常，在中期和后期形成多价染色体联结和滞后染色体，并产生与精子核相关的微核。NSF的资助还将使从各种研究项目获得的样本数字化成为可能。这将使有兴趣的科学家能够访问互联网使用公共数据库，从而提供一个无与伦比的机会，在本地和全球范围内共享信息。来自多个实验室的个人将远程执行程序，从而扩大研究范围，增加发表和传播结果的机会，同时最大限度地减少劳动力和动物数量。在直接的研究应用之后，教师和他们的学生将通过访问以前仅为少数选定的人保留的数据而获得授权。作为课堂练习访问的图像数据库将使学生接触到热门和令人兴奋的新研究，并培训他们使用21世纪的技术，从而促进科学发展，使他们具有竞争优势。