Combined Biophysical and Biochemical Study of Single Cells

描述（由申请人提供）：磁共振波谱（MRS）是医学科学中一种卓越的诊断方法，用于描绘化学成分、疾病和功能。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在内的小规模研究时。在这里，我们测试了一个假设，即在标准原子力显微镜（AFM）探针的悬臂部分制造的新型射频微线圈可以实现新的混合AFM/MRS技术。我们使用实验室开发的纳米制造工艺，在可偏转的AFM传感探针上创建平面射频线圈，从而允许高度聚焦光谱和纳米离散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)调查。探针将在具有已知相对性的水凝胶体系中进行评估。在目标2中，我们将确定？射频探针使单细胞的无创光谱。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尖端的挠度测量，这发生在尖端与样品接触时。该实验的成功完成有望为研究人员提供一种新的工具，该工具将与现有的诊断工具在形态学和形态学的定位精度方面竞争

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.