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

加州州立大学长滩的李博士和安德伍德博士获得了一笔赠款，用于购买一套成像系统，该系统由一台带有电动载物台的荧光显微镜、CCD摄像机、计算机和软件组成。 该系统将提供无偏差的体视学能力，以及1）可视化免疫荧光标记的样品; 2）产生三维（3-D）重建;和3）建立可通过互联网访问的图像数据库的能力。 李博士的研究目标是了解记忆形成和激素对斑胸草雀海马体神经发生的影响。 成年鸟类和哺乳动物的海马体保留了大量可塑性的能力，包括新细胞（细胞发生）和神经元（神经发生）的诞生，以应对许多环境影响，包括学习和记忆的形成。 李博士最近发现，海马细胞也是由于损伤而产生的，并聚集在斑胸草雀大脑受损部位周围。 受伤的大脑区域也富含芳香化酶，一种将睾丸激素转化为雌激素的酶。 雌激素，反过来，已知促进分化和神经元的存活。 这些有趣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神经胶质的因素。 目的是通过训练鸟类学习任务、损伤海马、注射BrdU（一种有丝分裂标记物）以及通过使用细胞化学技术评估细胞增殖、迁移和/或分化的模式来开发鸟类海马对环境影响的反应能力。 新的BrdU阳性细胞将被可视化，并使用MicrotechnologyField StereoInvestigator成像系统计数;双标记程序将被用于确定新细胞是神经元还是神经胶质。 安德伍德博士的研究目标是了解染色体异常、性别比例偏差和红蝴蝶“过剩”雄性进化之间的关系。 安德伍德博士的研究发现了斑蝶生殖细胞形成过程中染色体配对的异常。 一个亚种显示出26个二价体的正常补体;然而，另一个亚种在同一个体内从细胞到细胞显示出极端的变化。 异常亚种的初级性别比为75%雄性，而正常亚种的初级性别比为50%雄性。 蝴蝶具有染色体性别决定作用，精子发生异常，多价染色体联合，中、后期形成落后染色体，精子核伴随微核的产生。 国家科学基金会的资助也将使从各种研究项目中获得的样品的鉴定成为可能。 这将产生公共数据库，供感兴趣的科学家访问互联网，从而提供了一个无与伦比的机会，在当地和全球共享信息。来自多个实验室的个人将远程执行程序，从而扩大了研究范围，增加了结果出版和传播的机会，同时最大限度地减少了劳动力和动物数量。除了直接的研究应用，教师和他们的学生将通过访问以前只保留给少数人的数据而获得权力。 作为课堂练习访问的图像数据库将让学生接触热门且令人兴奋的新研究，并训练他们使用21世纪技术，从而推进科学并赋予他们竞争优势。

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.