SBIR-STTR Award

Locating fluorescently-tagged cells in whole organisms is important to the development of gene therapy, stem cell therapy delivery methods, and for drug research. This project will develop an intelligent microtome to identify fluorescently labeled cells in whole mice and to collect those cells in thin sections. During Phase I, a small-mammal cryomicrotome with LED-based tissue illumination for fluorescence imaging will be constructed. Several CCD cameras will be evaluated to establish the spatial resolution and gray-scale resolution needed to provide necessary fluorescence-imaging capabilities for the instrument. The instrument will be tested on isolated muscle, tissue homogenates containing green fluorescent protein (GFP) labeled cells, and on whole mice. The cells expressing GFP in these samples will provide data for the development of next-generation instrumentation during Phase II. Phase II will add capabilities to use the fluorescence image data to intelligently acquire thin sections only when fluorescent cells are detected. Fluorescence images acquired during sectioning will provide morphological guides to identify and isolate the same cells in mounted sections by laser-capture microdissection. Automated thin-tissue acquisition will be an added capability and will be integrated with intelligent fluorescence analysis to produce a completely automated instrument. The instrument will allow processing of whole animals to obtain thin sections for laser dissection and for biochemical and/or PCR post processing of select tissue regions.

Thesaurus Terms:
artificial intelligence, biomedical equipment, biomedical equipment development, fluorescence microscopy, molecular /cellular imaging, sectioning, three dimensional imaging /topography computer program /software, computer system hardware, muscle, muscular dystrophy, phantom model bioimaging /biomedical imaging, digital imaging, genetically modified animal, green fluorescent protein, laboratory mouse

Fluorescent tagging of cells using dyes, fluorescent proteins, and quantum dots forms the basis for many powerful analytical methods in modern biology. Locating these cells within entire animals is necessary to study cell delivery, differentiation and gene expression. The Phase II SBIR research is developing an intelligent microtome to provide the means to locate cells within an intact organ or organism and to capture thin sections containing target cells. This instrument will serially section whole organs or small mammals, record multiple wavelength fluorescence images from the tissue after removal of each section, analyze changes in fluorescent patterns in real time, and automatically collect thin sections from areas where fluorescently tagged cells are present. The compact instrument will mount on a typical laboratory bench. Instruments will be constructed for testing in university research laboratories studying gene therapy, stem cell behavior, pulmonary dysfunction, and gene transfer by bacterial transfection. A niche analysis of the commercial market for the intelligent microtome was performed during Phase I. The analysis indicated broad interest in this low-cost approach to three- dimensional imaging and the selective automated approach to collection of thin sections.

Public Health Relevance:
New biomedical methods are being designed to deliver DNA, RNA, or stem cells to targeted locations within the body to treat disease conditions. The research described in this Phase II SBIR proposal will manufacture intelligent microtomes, based upon prototype development studies conducted in Phase I. These new instruments will be evaluated in university research laboratories for their capability to section and image entire research animals, to locate stem cells or cells in which DNA or RNA is expressed, and to collect those cells in thin sections for further examination. This research will help the answer the questions "Which cells within the entire organism are affected by the gene or stem cell therapy?" and "What is going on at a biochemical level within these cells?"

Public Health Relevance:
PROJECT NARRATIVE New biomedical methods are being designed to deliver DNA, RNA, or stem cells to targeted locations within the body to treat disease conditions. The research described in this Phase II SBIR proposal will manufacture intelligent microtomes, based upon prototype development studies conducted in Phase I. These new instruments will be evaluated in university research laboratories for their capability to section and image entire research animals, to locate stem cells or cells in which DNA or RNA is expressed, and to collect those cells in thin sections for further examination. This research will help the answer the questions "Which cells within the entire organism are affected by the gene or stem cell therapy?" and "What is going on at a biochemical level within these cells?"

Project Terms:
3D image; Abscission; Affect; Algorithms; Anatomic; Anatomical Sciences; Anatomy; Animal Experimental Use; Animal Experimentation; Animal Research; Animals; Area; Automated Information Processing; Automatic Information Processing; Bacteria; Biochemical; Biology; Blood flow; Body Tissues; Brachydanio rerio; Cell Communication and Signaling; Cell Signaling; Cells; Collection; Computer Programs; Computer software; Coon's Technic; Coon's Technique; DNA; Danio rerio; Data; Data Set; Dataset; Deoxyribonucleic Acid; Detection; Development; Disease; Disorder; Dysfunction; Effectiveness; Electromagnetic, Laser; Environmental air flow; Evaluation; Excision; Exhibits; Extirpation; Florida; Fluorescence; Fluorescence Agents; Fluorescent Agents; Fluorescent Antibody Technic; Fluorescent Antibody Technique; Fluorescent Antinuclear Antibody Test; Fluorescent Dyes; Functional disorder; Funding; Gene Expression; Gene Products, RNA; Gene Transfer; Gene Transfer Clinical; Gene Transfer Procedure; Gene-Tx; Genes; Genetic Intervention; Goals; Histology; Image; Images, 3-D; Immunofluorescence Technic; Immunofluorescence Technique; Instrumentation, Other; Intervention, Genetic; Intracellular Communication and Signaling; Investigators; Label; Laboratories; Laboratory Research; Laboratory Study; Lasers; Location; Lung; Mammalia; Mammals; Mammals, General; Mammals, Mice; Marketing; Measurement; Measures; Medical Research; Method LOINC Axis 6; Methodology; Methods; Mice; Microdissection; Microtome - medical device; Microtomy; Molecular; Molecular Biology, Gene Therapy; Mother Cells; Murine; Mus; NIH; National Institutes of Health; National Institutes of Health (U.S.); Organ; Organism; Pattern; Phase; Physiologic; Physiological; Physiology; Physiopathology; Preparation; Procedures; Process; Processing, Automatic Information; Progenitor Cells; Programs (PT); Programs [Publication Type]; Proteins; Q-Dot; Quantum Dots; RNA; RNA, Non-Polyadenylated; Radiation, Laser; Regional Blood Flow; Removal; Research; Research Personnel; Researchers; Respiratory System, Lung; Ribonucleic Acid; SBIR; SBIRS (R43/44); Sampling; Science of Anatomy; Sepsis; Signal Transduction; Signal Transduction Systems; Signaling; Small Business Innovation Research; Small Business Innovation Research Grant; Software; Stem cells; Surface; Surgical Removal; Testing; Therapy, DNA; Thin Sectioning; Thin Sectionings; Three-Dimensional Image; Time; Tissues; Training; Transfection; United States National Institutes of Health; Universities; Ventilation; Washington; Work; Zebra Danio; Zebra Fish; Zebrafish; analytical method; anatomy; base; biological signal transduction; bloodstream infection; cell behavior; commercialization; computer program/software; computerized data processing; cost; data processing; design; designing; disease/disorder; drug development; fluorescence imaging; fluorescent antibody; fluorescent dye/probe; gene product; gene therapy; genetic therapy; imaging; instrument; instrumentation; interest; living system; medical schools; meetings; microtome; pathophysiology; programs; prototype; public health relevance; pulmonary; resection; sample collection; signal processing; specimen collection; stem cell therapy; success; transfer of a gene