Versatile system to select and expand individual or multiple adherent-type cells

用于选择和扩增单个或多个贴壁型细胞的多功能系统

• 批准号: 7671178
• 负责人: VASAN VENUGOPALAN
• 金额: $ 34.99万
• 依托单位: LIGHTWORKS OPTICS, INC.
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-05-01 至 2011-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7671178
• 关键词: Ablation; Adherent Culture; Adhesions; Air; Apoptosis; Area; Behavior; Biocompatible; Biological; Biological Assay; Biological Sciences; Biomedical Research; Biotechnology; Calibration; Cell Line; Cell Membrane Alteration; Cell Separation; Cell Survival; Cell physiology; Cell surface; Cells; Characteristics; Classification; Collecting Cell; Development; Disease Progression; Drug Industry; Event; Excision; Exposure to; Fluorescence Microscopy; Genomics; Glass; Growth; Height; Holography; Human Resources; Image; Individual; Industry; Interferometry; Lasers; Marketing; Mechanics; Mediating; Methodology; Methods; Microscope; Microscopy; Morphology; Necrosis; Optics; Patients; Pharmacologic Substance; Phase; Photography; Physiologic pulse; Plasma; Polymers; Polymethyl Methacrylate; Population; Positioning Attribute; Process; Proteomics; Research Personnel; Sampling; Side; Small Business Innovation Research Grant; Sorting - Cell Movement; Source; Staging; Stem cells; Stress; Surface; System; Techniques; Technology; Testing; Therapeutic Agents; Training; biological research; cell type; cost; design; genetic analysis; genetically modified cells; improved; instrument; interest; irradiation; laser capture microdissection; new technology; novel; pressure; prototype; public health relevance; time use

DESCRIPTION (provided by applicant): The ability to select, isolate, and expand single adherent-type cells or cell colonies is critical in many areas of biological research. Our objective is to develop a novel technology that integrates pulsed laser microbeam irradiation and polymer microdevices for single cell selection, isolation, and expansion. Cell selection and isolation are achieved by culturing adherent cells on the top of discrete polymer micropallet (30-250-5m sides, 30-100-5m height) fabricated on top of a glass surface that facilitates biological imaging. Once an adherent cell of interest is identified (e.g., via morphological analysis or fluorescence microscopy), a pulsed laser microbeam is delivered proximal to the interface between the polymer and the underlying glass coverslip to release the cell and micropallet. Once collected the cell can, for example, be subject to genomic/proteomic analysis or cultured to form a monoclonal cell population. We aim to design and build an apparatus that can be integrated with standard biological microscopy platforms and operated by general users in both biomedical research and pharmaceutical/biotechnology industries. During phase I of this SBIR application, we will (a) develop methods to optimize the laser microbeam irradiation parameters (e.g., wavelength, pulse duration, pulse energy, and numerical aperture) to release the polymer micropallets while minimizing cellular exposure to physical stresses associated with the release process, (b) verify that the chosen laser microbeam parameters facilitate cell selection with minimal loss of cell viability/function; and (c) design, build, and test the prototype instrument. PUBLIC HEALTH RELEVANCE: This SBIR Phase I project will develop a novel technology integrating the use of laser radiation and polymer microdevices to enable the identification, selection and recultivation of single adherent-type cells or cell colonies. This technology will be designed for use by biomedical researchers as well as the biotechnology and pharmaceutical industries. An important application of this technology is that it provides a cost-effective approach to the formation of homogeneous (monoclonal) cell populations with specific characteristics and thus enable a variety of activities connected with understanding cellular behavior and disease progression as well as the evaluating the efficacy of potential therapeutic agents.

描述(申请人提供)：选择、分离和扩大单个贴壁型细胞或细胞集落的能力在生物学研究的许多领域是至关重要的。我们的目标是开发一种集成脉冲激光微束照射和聚合物微器件的新技术，用于单细胞的选择、分离和扩增。细胞选择和分离是通过将贴壁细胞培养在离散的聚合物微托盘(边长30-250-5m，高30-100-5m)的顶部实现的，该托盘是在玻璃表面上制造的，便于生物成像。一旦识别出感兴趣的附着细胞(例如，通过形态分析或荧光显微镜)，脉冲激光微束被输送到聚合物和下面的玻璃盖片之间的界面附近，以释放细胞和微托盘。例如，一旦收集到细胞，就可以进行基因组/蛋白质组分析，或者进行培养以形成克隆细胞群体。我们的目标是设计和制造一种可与标准生物显微镜平台集成并可由生物医学研究和制药/生物技术行业的普通用户操作的仪器。在这项SBIR应用的第一阶段，我们将(A)开发各种方法来优化激光微束照射参数(例如，波长、脉冲持续时间、脉冲能量和数值孔径)，以释放聚合物微托盘，同时最大限度地减少细胞暴露在与释放过程相关的物理应力之下；(B)验证所选择的激光微束参数是否有助于选择细胞，同时将细胞活力/功能损失降至最低；以及(C)设计、制造和测试原型仪器。与公众健康相关：SBIR第一阶段项目将开发一种新技术，将激光辐射和聚合物微设备的使用相结合，以实现对单个贴壁型细胞或细胞克隆的识别、选择和再培养。这项技术将被设计为供生物医学研究人员以及生物技术和制药行业使用。这项技术的一个重要应用是，它提供了一种具有成本效益的方法来形成具有特定特征的同质(克隆)细胞群体，从而使与了解细胞行为和疾病进展以及评估潜在治疗剂的疗效相关的各种活动成为可能。