SBIR-STTR Award

Under microscope, it is easy to identify cells of specific morphological phenotypes or interacting with other cell types. But it is not trivial to pick the very cell out without submicron-precision mechanical instrument (micro pipetting or laser micro-dissection). To facilitate image based single cell isolation, we seek to develop an LCD aided selection under microscope (LASUM) technology that will allow us to pick single live cells without bulky instrumentation or tedious single cell handling skills. Also, as the capturing process is done by one single overall exposure step, the throughput of this technology will be greatly enhanced compared with single cell manipulating technologies. The proposed method can be used to select and isolate cells based on organelle translocation, chromatin morphology, phagocytosis, synapse formation, protein co-localization, and most importantly, cell-cell interactions. And we will complete this proof-of-concept study by implementing LASUM to the following areas: (a) phage enrichment on single cells or beads; (b) isolation of tumor-killing immune cell clones.

Public Health Relevance Statement:
Narrative Under microscope, it is easy to identify cells of specific morphological phenotypes or interacting with other cell types. But it is not trivial to pick the very cell out without submicron-precision mechanical instrument (micro pipetting or laser micro-dissection). To facilitate image based single cell isolation, we seek to develop an LCD aided selection under microscope (LASUM) technology that will allow us to pick single live cells without bulky instrumentation or tedious single cell handling skills.

Project Terms:
Area; Bacteriophages; base; Cell Communication; Cell Separation; cell type; Cells; Chromatin; Clone Cells; Erythrocytes; Excision; Image; Immune; instrument; instrumentation; Lasers; Mechanics; Methods; Microdissection; Microscope; Morphology; Organelles; Phage Display; Phagocytosis; Phenotype; Process; Proteins; skills; submicron; synaptogenesis; Technology; tumor

Characterizing both functional and genetic heterogeneity among a pool of cells remains a major scientific challenge in immunology, cancer research, neurobiology and developmental biology. Isolating cells with the same phenotype is key to understand such heterogeneity. We would like to develop a non-fluidic, unique technology, LCD aided selection under microscope (LASUM, Fig.1) to address this need. During the phase I of this project, we completed the alpha version of prototype design and demonstrated its utility in three proof-of-concept experiments: (a) blood cell removal; (b) phage enrichment on single beads; (c) isolation of tumor-killing immune cell clones. We would like to continue our effort to bring this technology to market, which will be low-cost, high throughput, debris resistant, image-based with operation simplicity as preparing a microscope slide or washing a microtiter plate. Besides the device, we will unify the fragmentated cell isolation market using two associated kits: Enrich-Live and Enrich- Seq. The successful commercialization of this device/kits will enable thousands of cell- biology labs and hospitals in manipulating single cells on a computer screen.

Public Health Relevance Statement:
Narrative We here propose the continued development of a cost-effective and biocompatible single cell isolation technology that utilizes the Liquid Crystal Display (LCD) with growing resolution to create a dynamic selection photomask (Fig.1), while the identity and classification of cells and cell-pairs are achieved by pattern-recognition algorithms. Cell isolation can be achieved in two ways: unwanted particle populations will be trapped by local photo-polymerization /gelation, the selected cells can be collected by elution; alternatively, cells of interest can be trapped in place, while unwanted cells/debris can be washed away. The method can be used to select and isolate cells based on organelle translocation, chromatin morphology, phagocytosis, synapse formation, protein co-localization, cell-cell interactions and other time-resolved cellular behaviors which cannot be achieved by current sorting platforms in a high-throughput manner. To make it commercially viable (<$10K production cost), several technical goals are critical: the proprietary error-tolerant algorithm of particle locating, the use of mass-produced components and the software portal for including external high-resolution images from third party platforms. To make the throughput comparable to common micro-fluidic sorters, optimized image pattern recognition algorithms and parallel computation tools will be implemented. Multiple applications will be used as test cases for the validation of device/kits during Phase II development.

Project Terms:
Address; Algorithms; Antibodies; anticancer research; B-Lymphocytes; Bacteriophages; base; Biological Assay; biomaterial compatibility; Blood Cells; cell behavior; Cell Communication; Cell Separation; Cells; Cellular biology; Chromatin; Classification; Clone Cells; commercialization; Computer software; computerized tools; Computers; cost; cost effective; Cytotoxic T-Lymphocytes; design; Development; Developmental Biology; Devices; Excision; experimental study; Genetic Heterogeneity; Goals; Heterogeneity; high resolution imaging; Hospitals; Image; Immune; Immunology; interest; liquid crystal; Methods; Microfluidics; Microscope; Morphology; Neoplasm Circulating Cells; Neurobiology; operation; Organelles; parallel computer; particle; Pattern Recognition; Phagocytosis; Phase; Phenotype; photopolymerization; Population; Production; Proteins; prototype; Resistance; Resolution; Slide; Sorting - Cell Movement; synaptogenesis; T-Lymphocyte; Technology; Testing; Time; tumor; Tumor Tissue; Validation