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

Project Summary ACISLLC proposes a conceptually novel cellular heterogeneity assaybased on confocal Ramanspectrometry and Biomolecular Component AnalysisBCAThe goal of Phase II is to implement a BCA toolbox in a commercial confocal microRaman instrumentfor quantitative assessment of cellular heterogeneity and potential quantitative classification ofcellular states based on macromolecular compositions in specific organellesa single cell assaydeveloped during the Phase IPhase II focuses on bringing this new device to the marketThis project is motivated by the factthat there is no currently any commercial toolwhichprovides direct probing of local biomolecular concentration in live cellsAt the same timethistool is extremely valuable for solving a number of problemswhich require determination ofquantitative markers for different cellular statesiin diseased cell populationcancer non cancerdifferent cancer stages etciiduring cell drug interactionandiiiinthe intracellular processesapoptosisproliferationdifferentiationetcThe Phase I research and development of this assay have met stated technical milestonesInparticularit is found that the BCA potential for characterization of cellular heterogeneitysignificantly exceeds that of the standard math tool of Principal Component Analysis routinely usedfor analysis of Raman spectraThe specific aims of this proposal areAdaptation of a Thermo Fisher micro Raman system DXRmodel to meet requirements for BCA applicationsDevelopment of Biomolecular Component AnalysisSoftware package in Cand or Java and incorporation to DXRmodel of Thermo Fisher micro RamansystemValidation of the BCA toolbox integrated with Thermo Fisher micro Raman system DXRmodel for ongoing biomedical researchaComparative analysis of heterogeneity between IDHmutated and WT IDH glioma cells and assessment of cell drug interaction in IDH mutant cells incollaboration with NCIBethesdaMDbMolecular profiling of cellular organelles forcategorization of Prostate Cancer with RPCIBuffaloNYPre market evaluation of theThermo Fisher DXRmicro Raman systemequipped by the BCA toolboxThe outcome of this project will be a commercial confocal micro Raman system with an implementedBCA toolboxalong with customized software for quantitative assessment of cellular heterogeneitywhich can be used for a broad range of biomedical applications in many sectors such as clinicallabsBiomedical cellular Research labsPharmaceutical industriesNational Cancer Institutesetc