Advanced Immunocytochemistry, In Situ Hybridization and Live Cell Imaging

描述(由申请人提供)： 高级免疫细胞化学、原位杂交和活细胞成像--2005年10月12日至10月25日 冷泉港实验室计划在2005-2009年秋季继续举办一门名为《高级免疫细胞化学、原位杂交和活细胞成像》(以前称为《高级原位杂交和免疫细胞化学》)的课程。这是一个为期两周的短期强化课程，培训学生(从研究生到首席调查人员)直接使用先进和/或专门的显微镜技术来定位核酸和蛋白质，并提供这些技术背后的生物化学概念的背景知识。学生通过免疫细胞化学和荧光原位杂交(FISH)获得定位基因、染色体、mRNAs和蛋白质的第一手经验，制备和使用标记的FISH核酸探针和标记抗体进行免疫定位，使用不同的抗原-抗体系统(荧光、酶、胶体金)进行实验，并使用GFP融合蛋白使用标准荧光、光漂白后的荧光恢复和共振能量转移来定位、定量和分析活细胞中的蛋白质。所有实验都使用了一系列最先进的显微镜方法(荧光显微镜、激光共聚焦扫描显微镜、去卷积显微镜)，学生有机会比较不同系统在不同应用中的使用情况。这些技术将用于利用光谱核型和比较基因组杂交(CGH)在DNA水平上表征癌基因和抑癌基因的拷贝数变化，并在固定细胞和活细胞中分析与人类致癌相关的mRNAs和蛋白质(如增殖标记物和抑癌基因产物)的细胞分布和表达状况。该课程由一支由该领域国际公认的领导者和一支敬业的、技术熟练的助手组成的团队授课，他们共同提供实践培训。讲师被邀请提供最新的报告，这些报告提供了学生在当前研究中学习的技术的使用实例。

DESCRIPTION (provided by applicant): Advanced Immunocytochemistry, In Situ Hybridization and Live Cell Imaging -- October 12- October 25, 2005 The Cold Spring Harbor Laboratory proposes to continue a course entitled "Advanced Immunocytochemistry, In Situ Hybridization and Live Cell Imaging" (previously called "Advanced In Situ Hybridization and Immunocytochemistry"), to be held in the Fall of 2005-2009. This is a short, two week, intensive course which trains students (ranging from graduate students to Principal Investigators) to directly use advanced and/or specialized techniques in microscopy to localize nucleic acids and proteins and provides background on the biochemical concepts underlying these techniques. Students gain first-hand experience in localizing genes, chromosomes, mRNAs and proteins, by immunocytochemistry and fluorescence in situ hybridization (FISH), preparing and using labeled nucleic acid probes for FISH and labeled antibodies for immunolocalization, performing experiments using different antigen-antibody systems (fluorescence, enzymatic, colloidal gold), and employing GFP-fusion proteins to localize, quantify, and analyze proteins in living cells using standard fluorescence, fluorescence recovery after photobleaching, and resonance energy transfer. All experiments are performed using a range of state-of-the-art microscope methods (epifluorescence microscopy, confocal laser scanning microscopy, deconvolution microscopy) and students have the opportunity to compare the use of different systems in different applications. The techniques will be used to characterize copy number changes of oncogenes and tumor suppressor genes on the DNA-level using spectral karyotyping and comparative genomic hybridization (CGH), and to analyze the cellular distribution and the expression status of mRNAs and proteins relevant to human carcinogenesis (such as proliferation markers and tumor suppressor gene products) as well as to the pathogenesis of other human diseases in fixed and living cells. The course is taught by a team of internationally recognized leaders in the field and a team of dedicated, technically adept assistants who together provide hands-on training. Lecturers are invited who give up-to-the-minute reports that provide examples of the use of techniques the students are learning in current research.