Combined Biophysical and Biochemical Study of Single Cells

描述(申请人提供)：磁共振波谱(MRS)是医学中描述化学成分、疾病和功能的一种卓越的诊断方法。更强大和更精确的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，特别是当它涉及包括细胞内光谱分析在内的小规模研究时。在这里，我们测试了一种假设，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AFM/MRS混合技术。我们使用我们实验室开发的纳米制造工艺，在可偏转的AFM传感探头上创建平面RF线圈，从而实现高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)探头的探测分辨率和性能。探头将在水凝胶体系中进行评估，并具有已知的相关性。在目标2中，我们将确定？RF探针实现单细胞非侵入性光谱分析的程度。我们将通过实施基于压电的反馈机制来对细胞系统进行MRS研究，以测量AFM尖端的挠度，该机制在尖端与样品接触时发生。拟议实验的成功完成有望为研究人员提供一种新的工具，在形态和组织结构的定位精度方面与现有的诊断工具竞争 化学数据，并能够在细胞层面上分析地下特征和生物物理调查。 与公众健康相关：我们的目标是通过开发一种用于单细胞实时光谱分析、生物物理和结构分析的新仪器工具，将高分辨率磁共振波谱(MRS)的生化分析能力与原子力显微镜(AFM)提供的空间定位功能和生物物理分析相结合。MRS是获取分子内化学和结构信息的一种卓越的分析方法。随着利用扩增技术检测生物标志物，结合AFM增强定位，MRS可以作为一种诊断工具，特别是随着先进的细胞内MRS重建技术的发展。

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.