MRI: ACQUISITION OF AN IMAGING SURFACE PLASMON RESONANCE SPECTROMETER FOR QUANTITATIVE ASSESSMENT OF SURFACE ADSORBING SPECIES

技术摘要表面等离子体共振(SPR)是一种在金属涂层表面进行的逝去波折射率测量，已被证明是一种定量、灵敏、无标记的技术，用于测量蛋白质、DNA、细胞和小分子与表面固定的捕捉剂的结合动力学。结合SPR成像(SPRI)，可以灵敏、准确地定量细胞面积和细胞-底物相互作用的动态变化，如膜褶皱。SPRI还提供了细胞蛋白质分泌的量化，以及表面蛋白质密度随时间和位置的变化。定制设计的SPRI将帮助研究人员研究多肽、纳米颗粒、蛋白质和其他生物偶联物在聚合物、自组装单分子层甚至培养中的活细胞上的吸附。荧光显微镜配置将能够捕获细胞形态信息、配体和受体的空间分布以及竞争性分子摄取研究，当这些研究与SPRI的吸附和动力学数据相结合时，将为定量和统计测量细胞材料相互作用提供机会。2009年，贝克尔博士与圣文森特/圣玛丽高中(STVM)开始了一项长期的外展工作。它已经成长为包括10名额外的UA教职员工。独一无二的是，每年，在每个年级，每个学生都被要求完成一个为期一年的探究性科学项目，作为科学课程的一部分。科学探究方法鼓励学生提出深入的问题，然后致力于寻找答案，包括进行实验，而不仅仅是记住事实。学生通过仔细和有条不紊地进行实验，与教师、研究生、高中教师和学生团队进行研究和咨询，来研究和确定他们着手解决或回答的问题。结果被表述成ACS聚合物预印本样式的手稿。在11个团队(学生、教师、教师)的共同努力下，STVM赢得了Harold C.Shaw杰出学校奖，并被评为2010年和2011年俄亥俄州杰出高中科学项目。我们的目标是使用学生在科学探究项目中收集的真实数据来演示折射率、光学吸附和蛋白质吸附的物理现象，这些现象将被纳入高中物理课程的新模块。非技术摘要分子与表面的相互作用在许多生物和物理过程中扮演着重要的角色。这些生物分子的吸附对于设计具有抗菌性能的材料、开发组织工程支架、骨生长和药物输送具有重要意义。成像表面等离子体共振仪(SPR)的获取将提供蛋白质、多肽和纳米颗粒在表面吸附的信息。阿克伦大学(UA)的这一新能力将为12个研究小组的学生和教职员工提供机会，研究生物材料领域的基本和应用问题。这种额外的能力将加速新生物材料的开发，并为研究生和本科生提供重要的培训。此外，参加加州大学为期一年的科学探究项目的当地高中生将使用该仪器来研究吸附动力学。我们的目标是使用学生收集的真实数据来演示折射率、光学吸附和蛋白质吸附的物理现象，这些现象将被纳入高中物理课程的新模块。

Technical AbstractSurface plasmon resonance (SPR) is an evanescent wave refractive index measurement made at a metal-coated surface and has been demonstrated to be a quantitative, sensitive, and label-free technique for measuring the binding kinetics of proteins, DNA, cells, and small molecules to surface-immobilized capture agents. The combination of imaging with SPR (SPRI) allows sensitive and accurate quantification of cell areas and dynamic changes in cell-substrate interactions such as membrane ruffling. SPRI also affords quantification of secretion of proteins from cells, and changes in surface protein density as a function of time and location. The custom-designed SPRI will aid researchers in studying adsorption of peptides, nanoparticles, proteins and other bioconjugates on polymers, self assembled monolayers and even live cells in culture. The fluorescence microscopy configuration will enable the capture of cell morphology information, spatial distributions of ligands and receptors and competitive molecular uptake studies which when coupled to the adsorption and kinetic data from SPRi will provide opportunities for quantitative and statistical measurements of cell material interactions. In 2009, Dr. Becker initiated a long-term outreach effort with St. Vincent / St. Mary High School (STVM). It has grown to include 10 additional faculty members at UA. Uniquely, EVERY year, at each grade level, EVERY student is required to complete a year-long, inquiry-based science project as part of the science curriculum. The science inquiry method encourages students to ask in depth questions and then to apply themselves, including conducting experiments, to find the answers, rather than merely memorize facts. The students research and identify a problem that they proceed to solve or answer by carefully and methodically performing experiments, conducting research and consulting with the teams of faculty, graduate students, high school teachers and students. Results are formulated into ACS POLYMER PREPRINT style manuscripts. Collectively the efforts of 11 teams (student, faculty, teacher) led to STVM in winning the Harold C. Shaw Outstanding School Award and named as the outstanding high school science program in the State of Ohio in both 2010 and 2011. We aim to use real data collected by the students in the science inquiry program to demonstrate physical phenomena of refractive index, optical adsorption and protein adsorption which will be incorporated into new modules in the high school physics curriculum.Non-Technical AbstractThe interactions of molecules with surfaces play an important role in many biological and physical processes. The adsorption of these biomolecules are important for designing materials that have antimicrobial properties, developing scaffolds for tissue engineering, bone growth, and drug delivery. The acquisition of Imaging Surface Plasmon Resonance Instrumentation (SPR) will provide information on adsorption of proteins, peptides and nanometer-size particles on surfaces. This new capability at the University of Akron (UA) will provide opportunities for students and faculty members in twelve research groups to investigate fundamental and applied problems in the area of biomaterials. This additional capability will accelerate the development of new biomaterials and provide important training to graduate and undergraduate students. In addition local high school students participating in a year-long science inquiry program at UA will use the instrumentation to study kinetics of adsorption. We aim to use real data collected by the students to demonstrate physical phenomena of refractive index, optical adsorption and protein adsorption which will be incorporated into new modules in the high school physics curriculum.