Acquisition of a Multi-User Scanning Laser Confocal for Science and Engineering Research at Hopkins University

拟采购一台具有双光子功能的扫描激光共聚焦显微镜，用于测量活细胞、组织、大分子组装体和新型材料的三维构象。有了这种新的显微镜，一个可以（i）切片干净，薄的光学切片厚荧光标本和光散射材料，（ii）查看平行于视线的标本平面，（iii）深入到光散射组织，活细胞，复杂的流体，和复杂的材料，（iv）获得非常高的分辨率的三维视图。这种新的显微镜将补充霍普金斯学院目前使用的实验技术，活跃在生物医学工程，化学工程，材料科学和生物学的研究项目。该提案详细介绍了新SLCM将产生巨大影响的四个项目。这些项目包括：（2）研究上皮细胞中膜蛋白的横向组织，强调糖基化磷脂酰肌醇锚定蛋白（GPI-蛋白）的包装，用于靶向递送到形态极化细胞的顶端表面;（3）研究染色质绝缘子影响增强子-启动子相互作用的分子机制，特别是研究位于果蝇gypsy反转录转座子5'转录非翻译区的gypsy绝缘子序列;（4）以高空间和时间分辨率监测介导细胞和微细胞器易位的过程。获得共聚焦显微镜的主要教育目标之一是训练霍普金斯的科学家和工程师使用生命和材料科学中最通用和最有前途的调查工具之一。 当然，如果没有适当的训练，这样一个强大的工具是无法正确使用的，也无法充分发挥其潜力。因此，两个合作- PIS将教一个新的课程，题为“实用共焦显微镜”将提供在1月的闭会期间，在科学和工程谁有兴趣学习如何使用共焦显微镜在他们的研究生研究的高级研究生。

It is proposed to purchase a scanning laser confocal microscope with two-photon capabilities to measure the 3D conformation of living cells, tissues, macromolecular assemblies, and novel materials. With this new microscope, one can (i) slice clean, thin optical sections out of thick fluorescent specimens and light scattering materials, (ii) view specimen planes parallel to the line of sight, (iii) penetrate deep into light-scattering tissues, living cells, complex fluids, and complex materials, and (iv) gain 3-D views at very high resolution. This new microscope will compliment the experimental techniques currently used by Hopkins faculty, active in research projects in biomedical engineering, chemical engineering, materials science and biology. This proposal details four projects where the new SLCM will have a dramatic impact. These projects include: (1) the development of a high resolution particle-tracking instrument coupled to the new confocal microscope to probe the in vitro and in vivo mechanical properties of biopolymers; (2) studying the lateral organization of membrane proteins in epithelial cells emphasizing the packaging of glycosylphos-phatidylinositol-anchored proteins, GPI-proteins, for targeted delivery to the apical surface of morphologically polarized cells; (3) studying the molecular mechanisms by which chromatin insulators affect enhancer-promoter interactions; in particular, studying the gypsy insulator, a sequence located in the 5' transcribed untranslated region of the gypsy retrotransposon of Drosophila; (4) monitoring the process of transfecting cells and micro-organelles translocations with high spatial and time resolution. One of the primary educational objectives for acquiring a confocal microscope is to train Hopkins scientists and engineers to use one of the most versatile and promising tools of investigation in the life and materials sciences. Of course, such a powerful instrument cannot be used properly and to its full potential with out proper training. Therefore, two of the Co- PIS will be teaching a new course entitled "Practical Confocal Microscopy" to be offered during the January intersession to advanced graduate students in science and engineering who are interested in learning how to use the confocal microscope in their graduate research.