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的尖端增强红外烧蚀（nanoIR-MS）的纳米级质谱成像新技术。b)将该技术应用于单细胞成像。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。鉴于目前的技术限制，目前大多数对生物系统的分析都是在细胞群上进行的，并假设该组的总体平均值将产生有用的结果。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循环中的阶段、历史和与环境的相互作用表现出不同的代谢组成。因此，为了检测罕见细胞（如循环肿瘤），对细胞进行单独分析是很重要的