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-13-140, is submitted by Advanced Cytometry Instrumentation Systems (ACIS), a company founded in 2001 to commercialize cutting-edge technologies via R&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 3D 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 1 : Optimization of confocal Raman micro-spectrometer design for multi-cycle long- term measurements of individual eukaryotic cells.
Aim 2 : Software and database development for semi-automatic Biomolecular Component Analysis.
Aim 3 : Validation of the proposed micro-Raman-BCA technology for single cell analysis and quantitative characterization of cellular heterogeneity.
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.
| Levchenko, Svitlana M; Kuzmin, Andrey N; Pliss, Artem et al. (2017) Macromolecular Profiling of Organelles in Normal Diploid and Cancer Cells. Anal Chem 89:10985-10990 |
| Kuzmin, Andrey N; Levchenko, Svitlana M; Pliss, Artem et al. (2017) Molecular profiling of single organelles for quantitative analysis of cellular heterogeneity. Sci Rep 7:6512 |
| Kuzmin, Andrey N; Pliss, Artem; Prasad, Paras N (2017) Ramanomics: New Omics Disciplines Using Micro Raman Spectrometry with Biomolecular Component Analysis for Molecular Profiling of Biological Structures. Biosensors (Basel) 7: |
