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.

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 正常细胞和癌细胞都将在PTM-TE探针存在下孵育，之后细胞将被回收，清洗，并重新悬浮在新鲜的培养液中。按照Acert人员的建议，细胞将被放置在一个扁平的细胞培养类型的样品容器中，放置在ESR显微镜单元中，并将获得单个细胞的ESRM图像。自旋密度的局域化将在亚细胞格子内确定。这些实验将使我们能够确定细胞对探针的摄取是否会导致细胞内的弥漫性浓度，或者PTM-TE探针是否定位于特定的亚细胞位置。如果发现是这种情况，那么就有可能对分子进行修饰，以便通过探针来引导特定的亚细胞靶向。 我们正在开发和表征Linc-BUO和其他嵌入生物兼容和透氧聚合物材料中的颗粒自旋探针。作为这些工作的延伸，我们建议使用三苯基分子探针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。我们实验室已经研制出了掺杂PTM-TE的PDMS，这些探针通常被称为“SpinChips”。我们将使用Acert的ESRM能力来帮助确定与这些材料一起使用的最佳掺杂浓度。ESRM将允许我们确定可能导致自旋-自旋相互作用和所获得信号的人为线宽的三苯基分子的聚集程度--这是应该避免的担忧。