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.
Thesaurus Terms: Ablation; Abscission; Adherent Culture; Adhesions; Air; Apoptosis; Apoptosis Pathway; Area; Assay; Behavior; Bioassay; Biocompatible; Biologic Assays; Biologic Sciences; Biological; Biological Assay; Biological Sciences; Biomedical Research; Biotechnology; Blood Plasma; Calibration; Cell Death, Programmed; Cell Function; Cell Isolation; Cell Line; Cell Lines, Strains; Cell Membrane Alteration; Cell Process; Cell Segregation; Cell Separation; Cell Separation Technology; Cell Survival; Cell Viability; Cell Physiology; Cell Surface; Cellline; Cells; Cellular Function; Cellular Physiology; Cellular Process; Characteristics; Collecting Cell; Development; Disease Progression; Drug Industry; Electromagnetic, Laser; Event; Excision; Exposure To; Extirpation; Fluorescence Microscopy; Generalized Growth; Genetic Analyses; Genomics; Glass; Growth; Height; Holography; Human Resources; Image; Individual; Industry; Industry, Pharmaceutic; Interferometry; Investigators; Lasers; Life Sciences; Manpower; Marketing; Mechanics; Mediating; Method Loinc Axis 6; Methodology; Methods; Methods And Techniques; Methods, Other; Microscope; Microscopy; Microscopy, Fluorescence; Microscopy, Light, Fluorescence; Monolayer Culture; Morphology; Mother Cells; Necrosis; Necrotic; Optics; Pmma; Patients; Pharmaceutical Agent; Pharmaceutical Industry; Pharmaceuticals; Pharmacologic Substance; Pharmacological Substance; Phase; Photography; Physiologic Pulse; Plasma; Poly(Methyl Methacrylate); Polymers; Polymethyl Methacrylate; Polymethylmetacrylate; Polymethylmethacrylate; Population; Position; Positioning Attribute; Pressure; Pressure- Physical Agent; Process; Progenitor Cells; Proteomics; Pulse; Radiation, Laser; Removal; Research Personnel; Researchers; Reticuloendothelial System, Serum, Plasma; Sbir; Sbirs (R43/44); Sampling; Serum, Plasma; Side; Small Business Innovation Research; Small Business Innovation Research Grant; Sorting - Cell Movement; Source; Staging; Stem Cells; Stress; Subcellular Process; Surface; Surgical Removal; System; System, Loinc Axis 4; Techniques; Technology; Testing; Therapeutic Agents; Tissue Growth; Training; Biological Research; Cell Sorting; Cell Type; Cost; Cultured Cell Line; Design; Designing; Genetic Analysis; Genetically Modified Cells; Imaging; Improved; Instrument; Interest; Irradiation; Laser Capture Microdissection; New Technology; Novel; Ontogeny; Personnel; Pressure; Prototype; Public Health Relevance; Resection; Sorting; Time Use
