ESR MICROSCOPY OF MOLECULAR PROBES IN CELLS AND POLYMERIC MATERIALS

细胞和聚合物材料中分子探针的 ESR 显微镜

• 批准号: 8364105
• 负责人: PERIANNAN KUPPUSAMY
• 金额: $ 0.05万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-01 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8364105
• 关键词: Biocompatible; Cancerous; Cell Culture Techniques; Cells; Culture Media; Diffuse; Fibroins; Funding; Grant; Human Resources; Image; Incubated; Individual; Location; Microscopy; Molecular Probes; National Center for Research Resources; Oxygen; Particulate; Principal Investigator; Research; Research Infrastructure; Resources; Sampling; Signal Transduction; Silk; Source; Technology; United States National Institutes of Health; cellular targeting; cost; density; polydimethylsiloxane; research study; uptake

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Both normal and cancerous cells will be incubated in the presence of PTM-TE probe, after which the cells will be recovered, washed, and re-suspended in fresh culture medium. The cells will be placed in a flat cell-culture type sample container, as recommended by ACERT personnel, placed in the ESR microscopy unit, and ESRM images of individual cells will be obtained. Localization of spin density will be determined within the sub-cellular compartments. These experiments will allow us to determine whether or not cellular uptake of the probes will result in diffuse concentration within the cells, or whether the PTM-TE probe localizes to a particular subcellular location. If this is found to be the case, it may then be possible to modify the molecules so as to direct specific sub-cellular targeting by the probes. We are developing and characterizing LiNc-BuO and other particulate spin-probes that are embedded in a biocompatible and oxygen-permeable polymeric material. As an extension of these efforts, we propose to use the trityl molecular probes PTM-TE and PTM-TC as dopants for oxygen-permeable polymeric materials, including polydimethylsiloxane (PDMS) and silk fibroin. PDMS doped with PTM-TE has been developed in our lab, and these probes are commonly referred to as "SpinChips." We shall use the ESRM capability of ACERT to assist in determining the optimal doping concentration for use with these materials. ESRM will permit us to determine the degree of aggregation of the trityl molecules that may result in spin-spin interaction and artificial line-broadening of the signal obtained - a concern that should be avoided.

该子项目是利用资源的众多研究子项目之一 由 NIH/NCRR 资助的中心拨款提供。子项目的主要支持 并且子项目的首席研究员可能是由其他来源提供的， 包括其他 NIH 来源。 子项目可能列出的总成本 代表子项目使用的中心基础设施的估计数量， NCRR 赠款不直接向子项目或子项目工作人员提供资金。 正常细胞和癌细胞都将在 PTM-TE 探针存在的情况下孵育，然后回收、洗涤细胞并重悬于新鲜培养基中。按照 ACERT 人员的建议，将细胞放置在扁平细胞培养型样品容器中，放置在 ESR 显微镜装置中，将获得单个细胞的 ESRM 图像。自旋密度的定位将在亚细胞区室内确定。这些实验将使我们能够确定探针的细胞摄取是否会导致细胞内的扩散浓度，或者 PTM-TE 探针是否定位于特定的亚细胞位置。如果发现情况确实如此，那么就有可能修改分子，以便通过探针指导特定的亚细胞靶向。 我们正在开发和表征 LiNc-BuO 和其他嵌入生物相容性和透氧聚合物材料中的颗粒自旋探针。作为这些努力的延伸，我们建议使用三苯甲基分子探针 PTM-TE 和 PTM-TC 作为透氧聚合物材料的掺杂剂，包括聚二甲基硅氧烷（PDMS）和丝素蛋白。我们的实验室已开发出掺杂有 PTM-TE 的 PDMS，这些探针通常称为“SpinChips”。我们将使用 ACERT 的 ESRM 功能来协助确定与这些材料一起使用的最佳掺杂浓度。 ESRM 将使我们能够确定三苯甲基分子的聚集程度，这可能会导致自旋-自旋相互作用和所获得信号的人为谱线展宽——这是一个应该避免的问题。