An Imaging Facility to Enhance Study of Cell and Molecular Biology

生物科学(61)米德尔伯里学院在基因组研究和微生物学方面的最新进展引起了人们对真核细胞的基因组修饰和这种修饰的表型效应的目视检查的兴趣。现在，学生们使用绿色荧光蛋白(GFP)“自然”标记野生型和突变型E2F转录因子，跟踪它们的定位，并在培养的哺乳动物细胞中监测它们的功能，这是一个新的扩展实验室项目，最近被改编并实施，作为真核细胞基因组修改的范例。他们在新建造的成像设备中使用配备了荧光和差分干涉对比光学的倒置显微镜来检查这些基因修饰的细胞。他们还使用低温恒温器和这种设施，包括带有荧光和DIC光学的直立显微镜，用于内分泌学、免疫学、发育生物学和细胞生物学的高级学习和课程研究项目。此外，提供这些具有数字捕获、显示和存储功能的显微镜，允许学生在每个工作站以小组形式工作，并鼓励他们在数据收集和分析方面积极合作。本项目中实施的程序改编自各种来源，包括科学文献(例如，Magae等人，1999；Kavaler等人，1999；Parfitt和Newman，1998)以及专著和技术手册(例如，Conn，1999；Lefkovits，1996)。这个项目的结果主要是通过分析学生在使用成像设备、荧光显微镜和DIC显微镜以及低温恒温器之前和之后准备的报告，以及通过跟踪学生在入学和毕业后的结果来评估的。

Biological Sciences (61) Recent growth in genomic research and microbiology at Middlebury College has generated interest in the genomic modification of eukaryotic cells and in the visual examination of the phenotypic effects of such modification. Students now use green fluorescent protein (GFP) to "naturally" tag wild- type and mutant E2F transcription factors, follow their localization and monitor their function in cultured mammalian cells, in an extended laboratory project newly adapted and implemented as a paradigm for the genomic modification of eukaryotic cells. They examine these genomically modified cells using inverted microscopes equipped for fluorescence and differential interference contrast optics in a newly constructed Imaging Facility. They also use a cryostat and this facility, including upright microscopes with fluorescence and DIC optics, for advanced study and course research projects in endocrinology, immunology, and developmental biology as well as cell biology. Additionally, provision of these microscopes with digital capturing, display and storage capabilities allows students to work at each work station in small groups and encourages their active collaboration in data collection and analysis. Procedures implemented in this project are adapted from various sources, including the scientific literature (e. g., Magae et al., 1999; Kavaler et al., 1999; Parfitt and Newman, 1998) and monographs and technical manuals (e. g., Conn, 1999; Lefkovits, 1996). The results of this project are evaluated primarily through the analysis of portfolios of student reports prepared before and after they used the Imaging Facility, fluorescence and DIC microscopes, and cryostat and by tracking student outcomes during matriculation and following graduation.