Single Cell chemical Imaging via nanoscale IR ablation - Mass Spectromety

通过纳米级红外烧蚀进行单细胞化学成像 - 质谱

基本信息

批准号：
8523513
负责人：
Kermit King Murray
金额：
$ 15.16万
依托单位：
ANASYS INSTRUMENTS CORPORATION
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-05-01 至 2014-12-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8523513
关键词：
Ablation American Type Culture Collection Anatomy Biochemical Biocompatible Materials Biology Biopsy Blood Brain Breast Cancer Cell Buffers Cancer Cell Growth Cell Culture Techniques Cells Chemicals Citrates Coagulation Process Collection Colon Colorado Culture Media Deposition Disease Eagles Environment Epithelial Exhibits Fluorescein Fluorescence Freezing Gene Expression Genomics Glucose Goals Harvest Heterogeneity Image Imagery Imaging Techniques Individual Ions Letters Light Liquid substance Lung MCF7 cell Malignant Neoplasms Mammalian Cell Mass Spectrum Analysis Membrane Metabolic Metabolism Metastatic Lesion Microscope Molecular Molecular Weight Morbidity - disease rate Mus Neoplasm Circulating Cells Neoplasm Metastasis Organelles Oryctolagus cuniculus PKH67 Pancreas Phase Phosphate Buffer Pituitary Gland Procedures Prostate Proteins Proteomics Recording of previous events Research Resolution Saline Sampling Scanning Probe Microscopes Schools Serum Slide Sodium Bicarbonate Solid Solutions Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization Spectrometry, Mass, Secondary Ion Staining method Stains Surgeon Suspension substance Suspensions System Techniques Technology Trypsin United States National Institutes of Health Veterinary Medicine Whole Blood base biological systems brain cell brain tissue cellular imaging fetal bovine serum innovation ionization ionization technique malignant breast neoplasm mass spectrometer metabolomics molecular mass nanoscale neoplastic cell oncology prevent programs public health relevance single cell analysis small molecule sodium citrate tool

项目摘要

DESCRIPTION (provided by applicant): The goals of this project are twofold: a) to develop a new technique for nanoscale Mass spectrometry imaging based on AFM based tip enhanced IR ablation (nanoIR-MS). b) to apply this technique towards the application of Single cell imaging. Information on the chemical composition within a cell has implications in the understanding of cell metabolism, division, disease states, ecological effects etc. Given current technology limitations, most current analyses of biological systems are performed on groups of cells with the assumption that an ensemble average from the group will yield a useful result. However, this typically is not a valid assumption as cells of the same type exhibit diverse metabolic makeup depending on their phase in the cycle, history and interaction with the environment. Thus it is important that cells be analyzed individually in order to detect rare cells (e.g. circulating tumor cells), transient cell states, the influence of the cell environment on cells and states and aid in the understanding of differences in gene expression, protein levels, and small- molecule distributions at the single cell level. Cell heterogeneity is particularly significant in the "-omis" fields such as genomics, proteomics, lipidomics, and metabolomics that characterize biological systems at a molecular level. This significance led to the NIH launching a special focus program on Single cell Analysis Tools in late 2011. The size of mammalian cells is on the order of 10 ¿m and therefore the imaging of single cells requires imaging spatial resolution of at least 1¿m. The nanoIR-MS technology has a potential spatial resolution of at least 10x better than this or 100 nm which offers the possibility of the imaging of biomolecules in organelles. But to achieve an innovative and commercially successful product from this proposal, 1 ¿m spatial resolution would suffice. One of the Specific Aims of this proposal is to demonstrate that the nanoIR-MS technique can be applied for Single Cell Imaging. We will demonstrate this on two types of Single cells: Cells from Mouse brain and also to identify single Circulating Tumor cells (CTCs). As reiterated in our Letter of Support from our collaborator, Prof. Yeh who is an Oncology research surgeon, CTCs are the fundamental entities primarily responsible for spawning metastatic disease and there is a current lack of characterization technologies to identify them . To cure epithelial-based cancers-such as cancers of the breast, prostate, lung, colon and pancreas-therapies need to be directed towards those cells that cause metastases. However, the majority of metastatic lesions are never biopsied due to anatomic inaccessibility or associated morbidity of the procedure. CTCs offer a readily accessible means of studying the biology of metastatic cells throughout the course of disease and are often referred to as 'Liquid Biopsy'.

描述（由申请人提供）：该项目的目标有两个：a）开发一种基于 AFM 的尖端增强红外消融（nanoIR-MS）的纳米级质谱成像新技术 b）将该技术应用于实际应用。关于细胞内化学成分的信息对理解细胞代谢、分裂、疾病状态、生态效应等具有重要意义。鉴于当前技术的限制，大多数当前的生物系统分析都是在具有以下特征的细胞群上进行的：假设然而，这通常不是一个有效的假设，因为同一类型的细胞根据其在周期中的阶段、历史和与环境的相互作用而表现出不同的代谢组成，因此这一点很重要。对细胞进行单独分析以检测稀有细胞（例如循环肿瘤细胞）、瞬时细胞状态、细胞环境对细胞和状态的影响以及帮助在单细胞水平上了解基因表达、蛋白质水平和小分子分布的差异在基因组学、蛋白质组学、脂质组学和代谢组学等表征生物系统的“-omis”领域尤其重要。这种重要性导致 NIH 在 2011 年底启动了一项关于单细胞分析工具的特别重点计划。哺乳动物细胞的大小约为 10 ¿ m，因此单细胞成像需要至少 1¿ m. nanoIR-MS 技术的潜在空间分辨率至少比此高 10 倍或 100 nm，这提供了细胞器中生物分子成像的可能性，但要从该提案中获得创新且商业上成功的产品，1 ¿ m 空间分辨率就足够了该提案的具体目标之一是证明 nanoIR-MS 技术可以应用于单细胞成像，我们将在两种类型的单细胞上进行演示：来自小鼠大脑的细胞，并用于识别。正如肿瘤研究外科医生 Yeh 教授在我们的合作实体的支持信中所重申的那样，CTC 是产生转移性疾病的主要原因，并且目前存在一个问题。缺乏鉴定技术来治疗基于上皮的癌症，例如乳腺癌、前列腺癌、肺癌、结肠癌和胰腺癌，治疗需要针对那些引起转移的细胞。由于解剖学上的难以接近或手术相关的发病率而从未进行过活检，CTC 提供了一种在整个疾病过程中研究转移细胞生物学的便捷方法，通常被称为“液体”。活检'。