Combined Biophysical and Biochemical Study of Single Cells

单细胞的生物物理和生化联合研究

• 批准号: 8414058
• 负责人: Corey P Neu
• 金额: $ 18.69万
• 依托单位: PURDUE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-05 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8414058
• 关键词: Advanced Development; Aging; Arthritis; Atomic Force Microscopy; Basic Science; Biochemical; Biocompatible Materials; Biological; Biological Markers; Biological Process; Biomedical Research; Caliber; Cell Culture System; Cell physiology; Cells; Cellular Structures; Cellular biology; Chemicals; Chondrocytes; Complement; Coupled; Data; Detection; Development; Diagnostic; Diagnostic Procedure; Dimensions; Disease; Electronics; Embryonic Development; Employment; Environment; Feedback; Goals; Growth; Heterogeneity; Hybrids; Hydrogels; Image; Imagery; Interferometry; Investigation; Knowledge; Laboratories; Lasers; Life; Magnetic Resonance; Magnetic Resonance Imaging; Magnetic Resonance Spectroscopy; Measurement; Measures; Medical; Microscopy; Monitor; Nuclear Magnetic Resonance; Outcome; Performance; Population; Principal Investigator; Process; Radial; Research; Research Personnel; Resolution; Sampling; Science; Signal Transduction; Spectrum Analysis; Speed; Staging; System; Techniques; Technology; Testing; Time; Work; analytical method; articular cartilage; base; cantilever; cartilage cell; cell injury; cell type; disease diagnosis; innovation; instrumentation; nanofabrication; nanometer; nanoscale; new technology; novel; operation; programs; radiofrequency; reconstruction; research study; single cell analysis; tool

DESCRIPTION (provided by applicant): Magnetic resonance spectroscopy (MRS) is a preeminent diagnostic method in medical science for depicting chemical composition, disease, and function. The advancement of more powerful and precise MRS instrumentation is critical to broaden the applicability of the technology to biomedical research and disease diagnosis, especially as it relates to small-scale studies including in-cell spectroscopy. Here, we test the hypothesis that novel radiofrequency microcoils fabricated on the cantilever portion of standard atomic force microscopy (AFM) probes can enable a new hybrid AFM/MRS technology. We use nanofabrication processes developed in our laboratory to create planar RF coils formed on a deflectable AFM sensing probe, therefore allowing for highly-focused spectroscopy and nanometer-discretized cell localization and biophysical analysis. We will pursue two Aims. In Aim 1, we will optimize the detection resolution and performance of AFM-coupled micro radiofrequency (?RF) probes. Probes will be evaluated in hydrogel systems with known relativity. In Aim 2, we will determine the extent that ?RF probes enable noninvasive spectroscopy of single cells. We will conduct MRS studies on cell systems through the implementation of a piezoelectric-based feedback mechanism for the deflection measurement of the AFM tip, which takes place when the tip is brought into contact with samples. Successful completion of the proposed experiments is expected to provide researchers with a new tool that will compete with existing diagnostic tools in terms of localization precision of morphological and chemical data and enable analysis of subsurface features and biophysical investigation in the cellular level. PUBLIC HEALTH RELEVANCE: We aim to complement the biochemical analysis capabilities of high-resolution magnetic resonance spectroscopy (MRS) with the spatial localization features and biophysical analysis provided by atomic force microscopy (AFM), through the development of a new instrumentation tool for real-time spectroscopy, biophysical, and structural analysis of single cells. MRS is a preeminent analytical method for the acquisition of chemical and structural information within molecules. With the detection of biomarkers through the employment of amplification techniques, combined with AFM-enhanced localization, MRS can be used as a diagnostic tool, especially with the development of advanced in-cell MRS reconstruction techniques.

描述（由申请人提供）：磁共振波谱（MRS）是医学科学中用于描述化学成分、疾病和功能的卓越诊断方法。更强大和精确的MRS仪器的进步对于扩大该技术在生物医学研究和疾病诊断中的适用性至关重要，特别是当它涉及到包括细胞内光谱学在内的小规模研究时。在这里，我们测试的假设，新的射频微线圈上制造的悬臂部分的标准原子力显微镜（AFM）探针可以使一个新的混合AFM/MRS技术。我们使用我们实验室开发的纳米纤维工艺来创建在可偏转的AFM传感探针上形成的平面RF线圈，因此允许高度聚焦的光谱和纳米离散化的细胞定位和生物物理分析。我们将追求两个目标。在目标1中，我们将优化AFM耦合微射频（？RF）探针。将在具有已知相关性的水凝胶系统中评价探针。在目标2中，我们将确定？RF探针能够对单细胞进行无创光谱分析。我们将通过实施基于压电的反馈机制对细胞系统进行MRS研究，该机制用于AFM针尖的偏转测量，当针尖与样品接触时发生。成功完成拟议的实验，预计将为研究人员提供一种新的工具，将与现有的诊断工具，在定位精度的形态和 化学数据，并能够在细胞水平上分析地下特征和生物物理研究。 公共卫生相关性：我们的目标是通过开发一种新的仪器工具，用于单细胞的实时光谱、生物物理和结构分析，利用原子力显微镜（AFM）提供的空间定位功能和生物物理分析，补充高分辨率磁共振光谱（MRS）的生化分析能力。MRS是一种用于获取分子内化学和结构信息的卓越分析方法。通过采用扩增技术检测生物标志物，结合AFM增强定位，MRS可用作诊断工具，特别是随着先进的细胞内MRS重建技术的发展。