SBIR-STTR Award

Tissue heterogeneity has always limited the information obtainable from genomic or proteomic analysis of biological samples in the study of disease. Tissue microdissection and cell sorting technologies have advanced tremendously over the last decade from simple manual tissue dissection to sophisticated laser capture microdissecting (LCM) instruments and high speed fluorescence assisted cell sorting systems (FACS). In combination with genomics and proteomics technologies it is now possible to generate cell specific transcriptome/proteome data advancing the identification of disease biomarkers and novel therapeutic targets. Currently, LCM and FACS are two main technologies for isolation of specific tissues and cell types. However, it is clear that these technologies are not sufficient to fully support the growing need for cell specific molecular data. This process is hampered by extremely high costs of LCM and FACS machines, high cost of their maintenance and their sophisticated interface. Further, each of them has specific limitations: 1) LCM performs cell and tissue collection using fixed tissues that often affects the quality of biological material such as RNA; 2) FACS requires fluorescent labels and works with dissociated cells. The need for the low-cost, reliable and simple-to-use instrument that would offer capabilities similar to LCM and FACS is evident. Here, we propose a novel capillary-based vacuum-assisted cell and tissue acquisition system (CTAS) that is envisioned as an attachment to existing inverted microscopes and will allow specific cell and cell cluster collection from fresh, fresh frozen and fixed tissues for downstream genomics and proteomics applications. This instrument is estimated to be in the price range of $5,000, therefore it will be affordable for any laboratory. This machine will have interchangeable key components and will be user friendly. Cell specific sorting/capture technology is a prerequisite for precise characterization of the specific cell types for understanding their function, regulation of the metabolism and drug development. Currently two major approaches for acquisition of specific cells are available: fluorescence assisted cell sorting (FACS) and laser-capture microdissection (LCM). These technologies are sophisticated, have high cost of maintenance and the instruments are very expensive. We are developing a low-cost vacuum-assisted capillary-based cell and tissue acquisition system (CTAS). It is a simple, non-invasive (unlike LCM it does not require tissue fixing and drying) technology that can be easily automated and offers wide range of cell- and tissue-specific separation parameters. In addition, it is estimated to be at least 20 times cheaper than any existing devices such as LCM or FACS. This simple, rapid, microcapillary-based technique will be capable of isolating cells and tissues from animal and human material for further downstream molecular analysis without compromising the preservation of macromolecules in the tissue. Low-cost CTAS featuring open system architecture and interchangeable components will play a key role in collection and processing of cell-specific genomics and proteomics data in biomedical experiments

Tissue heterogeneity is a serious limiting factor for sound cell-specific molecular studies of the disease including genomic or proteomic analysis. Tissue microdissection and cell sorting technologies have advanced tremendously over the last decade from simple manual tissue dissection to sophisticated laser capture microdissecting (LCM) instruments and high speed fluorescence assisted cell sorting systems (FACS). In combination with genomics and proteomics technologies it is now possible to generate cell specific transcriptome/proteome data, advancing the identification of disease biomarkers and novel therapeutic targets. Currently, LCM and FACS are the two main technologies for the isolation of specific tissues and cell types. However, due to their high costs and often sophisticated interface, these technologies are not sufficient to fully support the growing need for cell specific molecular data. Therefore, there is a tremendous need for a low-cost and simple-to-use microdissection device that would offer capabilities similar to LCM and FACS. The overall goal of this SBIR project is to develop a new low-cost microdissection instrument with cellular resolution. In phase I of this project we proposed to build a prototype and test the feasibility of a novel capillary- based vacuum-assisted cell and tissue acquisition system (CTAS) that was envisioned as an attachment to inverted microscopes. The proposed CTAS would be able to dissect tissues at cellular resolution and collect material (RNA or protein) for downstream applications (e.g. expression microarrays). Phase I of this project was highly successful. We developed a fully functional prototype and demonstrated its use for collection of specific cell types from mouse central nervous system (spinal cord and brain). The architecture and major components of CTAS, including the capillary holder, collector, vacuum source, CTAS holder and light source, were developed, tested and optimized. Phase II specific aims include 1) further development of the critical components of CTAS; 2) development of control unit and adjustable parameters; 3) further testing of CTAS on tissue sections; and cell cultures. In addition, the prototype will be tested in different laboratory settings including tissue dissection and cell specific collection from heterogeneous cell culture sources. NeuroInDx will complete this work, which will be necessary to successfully evaluate proposed CTAS, and will commercialize the instrument in phase III of this project.

Public Health Relevance:
Cell specific sorting/capture technology is a prerequisite for precise characterization of the specific cell types for understanding their function and regulation of the metabolism, as well as for preclinical translational research. Currently two major approaches for the acquisition of specific cells are available: fluorescence assisted cell sorting (FACS) and laser-capture microdissection (LCM). These technologies are sophisticated and the instruments are not only very expensive but have high maintenance costs. In phase I of this project, we developed a low-cost vacuum-assisted capillary-based cell and tissue acquisition system (CTAS) and demonstrated its feasibility and applicability for tissue microdissection and downstream applications. It is a simple, non-invasive (unlike LCM it does not require tissue fixing and drying) technology that can be easily automated and offers a wide range of cell- and tissue-specific separation parameters. We estimate that CTAS will be at least 5-10 times cheaper than LCM or FACS instruments. In phase II of this SBIR application, we propose further development of the instrument, its optimization and testing for the range of applications including tissue microdissection and cell specific collection from heterogeneous cell cultures. As part of Phase II, beta testing of CTAS will be carried out in several sites including academic laboratories and industry. This work will result in its full commercialization in the following phase III. This low-cost microdissection instrument will be affordable for virtually any research laboratory, and therefore, the demand will likely be very high given the growing need for cell specific analysis.

Thesaurus Terms:
Animal Experimental Use; Animal Experimentation; Animal Research; Architecture; Area; Basic Research; Basic Science; Biopsy; Blood Capillaries; Body Tissues; Brain; Budgets; Capillaries; Capillary; Capillary, Unspecified; Cell Components; Cell Culture Techniques; Cell Isolation; Cell Segregation; Cell Separation; Cell Separation Technology; Cell Structure; Cells; Cellular Morphology; Cellular Structures; Central Nervous System; Collection; Computer Programs; Computer Software; Dissec; Data; Development; Device Or Instrument Development; Devices; Disease; Disorder; Dissection; Electromagnetic, Laser; Electronics; Encephalon; Encephalons; Engineering / Architecture; Ensure; Fluorescence; Freezing; Gene Expression; Gene Expression Profile; Gene Products, Rna; Genomics; Genotype; Goals; Heterogeneity; Human Resources; Industry; Institution; Intermediary Metabolism; Label; Laboratories; Laboratory Research; Lasers; Life; Light; Metbl; Maintenance; Maintenances; Mammals, Mice; Manpower; Manuals; Marketing; Medical; Medulla Spinalis; Metabolic Processes; Metabolism; Mice; Microdissection; Microscope; Modeling; Molecular; Monitor; Murine; Mus; Nervous System, Brain; Nervous System, Cns; Neuraxis; Nucleic Acids; Operation; Operative Procedures; Operative Surgical Procedures; Performance; Pharmaceutical Agent; Pharmaceuticals; Pharmacologic Substance; Pharmacological Substance; Phase; Photoradiation; Price; Process; Proteins; Proteome; Proteomics; Rna; Rna, Non-Polyadenylated; Radiation, Laser; Regulation; Research; Research Specimen; Resolution; Ribonucleic Acid; Sbir; Sbirs (R43/44); Sampling; Site; Small Business Innovation Research; Small Business Innovation Research Grant; Software; Sorting - Cell Movement; Sound; Sound - Physical Agent; Source; Specimen; Speed; Speed (Motion); Spinal Cord; Surgical; Surgical Interventions; Surgical Procedure; System; System, Loinc Axis 4; Technology; Testing; Time; Tissue Banks; Tissue Collection; Tissue/Specimen Collection; Tissues; Training; Translational Research; Translational Research Enterprise; Translational Science; Vacuum; Work; Base; Biomarker; Capillary; Cell Culture Collection; Cell Morphology; Cell Sorting; Cell Type; Commercialization; Computer Program/Software; Cost; Device Development; Disease/Disorder; Flexibility; Gel Electrophoresis; Gene Expression Signature; Gene Product; Improved; Instrument; Instrument Development; Interest; Laser Capture Microdissection; New Therapeutic Target; Novel; Personnel; Pre-Clinical; Preclinical; Pricing; Prototype; Public Health Relevance; Sample Collection; Sorting; Sound; Specimen Collection; Surgery; Tissue Fixing; Tissue/Cell Culture; Tool; Transcriptome; Translation Research Enterprise; User-Friendly