Novel single cell assay for quantitative analysis of cell heterogeneity by noninvasive probing of molecular composition of specific organelles in individual cells

DESCRIPTION provided by applicant Novel single cell assay for quantitative analysis of cell heterogeneity by noninvasive probing of molecular composition of specific organelles in individual cells Project Summary This current SBIR application in response to PA is submitted by Advanced Cytometry Instrumentation Systems ACIS a company founded in to commercialize cutting edge technologies via Randamp D in the fields of nanotechnology photonics and bio photonics The current proposal is specifically focused on development and commercialization of next generation single cell assay for quantitative analysis of heterogeneity of eukaryotic cells by noninvasive probing of molecular composition in individual cells and or specific organelles This technology will enable real time monitoring of the concentrations of major classes of macromolecules proteins lipids DNA RNA as well secondary structures of proteins in regions of interests in live cells The technology proposed here integrates i highly sensitive high D resolution low phototoxic miniaturized and robust confocal Raman micro spectrometry tool designed to work in both academic and clinical laboratory and ii novel concept of user friendly Biomolecular Component Analysis BCA of measured data sets to generate time sequenced biomolecular composition profiles in individual cells Based on the above goals the following specific aims are proposed for this Phase I study Aim Optimization of confocal Raman micro spectrometer design for multi cycle long term measurements of individual eukaryotic cells Aim Software and database development for semi automatic Biomolecular Component Analysis Aim Validation of the proposed micro Raman BCA technology for single cell analysis and quantitative characterization of cellular heterogeneity PUBLIC HEALTH RELEVANCE We propose identification of cellular heterogeneity by time sequential profiling of subcellular sites of interests e g organelles macromolecular clusters fr their macromolecular content Our approach integrates i confocal Raman micro spectrometry setup modified to enable noninvasive low phototoxic real time probing of individual live cells and ii novel concept of semi automated Biomolecular Component Analysis of acquired data to generate time sequenced biomolecular profiles in individual cells