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

• 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
• 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

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（由应用程序提供）：该项目的目标是双重的：a）基于基于AFM的TIP增强IR消融（Nanoir-MS）开发纳米级质谱成像的新技术。 b）将此技术应用于单细胞成像的应用。细胞内化学成分的信息对理解细胞代谢，分裂，疾病状态，生态影响等有影响，鉴于当前的技术局限性，对生物系统的大多数分析都是对细胞组进行的，并假设该组的组合平均值将产生有用的结果。但是，这通常不是一个有效的假设，因为相同类型的细胞根据周期，历史和与环境相互作用的相位表现出潜水的代谢化妆。重要的是要单独分析细胞以检测稀有细胞（例如循环肿瘤 细胞），瞬态细胞状态，细胞环境对细胞和状态的影响，并有助于 在单细胞水平上对基因表达，蛋白水平和小分子分布的差异的理解。细胞异质性在“ -OMIS”领域（例如基因组学，蛋白质，脂肪组学和代谢组学）在分子水平上表征生物系统的特征尤其重要。这种意义导致NIH在2011年末启动了对单细胞分析工具的特殊焦点程序。哺乳动物细胞的大小为10€m，因此，单细胞的成像需要成像至少1€m的空间分辨率。 Nanoir-MS技术的潜在空间分辨率至少比其或100 nm好10倍，这为细胞器中生物分子的成像提供了可能。但是，为了从该提案中获得创新且在商业上成功的产品，1`空间分辨率就足够了。该提案的具体目的之一是证明纳米MS技术可以用于单细胞成像。我们将在两种类型的单细胞上证明这一点：小鼠大脑的细胞，并鉴定单个循环肿瘤细胞（CTC）。正如我们合作者YEH教授是肿瘤研究外科医生YEH教授的支持书中所重申的那样，CTC是首先负责产生转移性疾病的基本实体，目前缺乏识别它们的特征技术。为了治愈基于上皮的癌症，例如乳腺癌，前列腺，肺，结肠和胰腺切割，需要针对那些引起转移酶的细胞。但是，由于解剖学上的难以接近或该过程的相关发病率，大多数转移性病变永远不会被活检。 CTC在整个疾病过程中提供了一种容易获得的研究转移细胞生物学的方法，通常被称为“液体活检”。