Automated Isolation of Single Cells Using High-Resolution and Subcellular Imaging

PROJECT SUMMARYPairingdetailedphenotypiccharacterizationofcellswithdownstreammolecularanalysisrepresentsasignificant challengeespecially at the level of single cellsCell Microsystems has developed and commercializedthe CellRaftSystemwhich is currently the only commercially available means of imaging cells using brightfieldand or multichannel fluorescence prior to single cell isolation and downstream molecular analysisInvestigatorsinterested in pairing phenotypic data with genomic analysis at the single cell level have employed the CellRaftSystem for a range of innovative studies including stem cell differentiationtumor cell physiology and screeningof neuronsHoweverthe materials used in the currently commercially available CellRaft Arrays are not optimizedfor imaging quality and were selected for other properties such as the resealing behavior of the elastomer and cellculture compatibility of the plasticDuring the Phase I programalternative materials for the CellRaft Array wereexplored for improved optical properties such as refractive index agreement and reduced autofluorescenceOneparticularalternativematerialscombinationexhibitedfavorableperformanceinbothcriterialeadingtodramaticallyimprovedcellimagingperformanceHerewerefertothisversionoftheproductastheHighResolutionorHRCellRaftThefocusofourPhaseIIproposalistocommercializetheHRCellRaftforusersinterestedinpairingdetailedphenotypicimagingnotpossibleonthecurrentCellRaftArraye gsubcellularproteinlocalizationorganellephysiologyandsemiquantitativefluorescentsignalsaswellasautomatetheCellRaftSystemworkflowInaparallelprogramwearedevelopingahighlyautomatedinstrumenttheAutomatedIsolationandRetrievalAIRSystemforsinglecellimagingandisolationemployingthecurrentCellRaft ArrayDuring Phase II we will implement several design revisions to the optical capabilities of the AIRSystemaswellasitssoftwarebasedimageanalysisalgorithmstobetterleveragetheHRCellRaft simprovedopticalpropertiesThenewproposedinstrumentisreferredtohereastheCLEARAIRSystemAdditionalfluorescence detection channelssix instead of three in the AIRSystemhigher magnification objectivesXX andX as opposed to the single objectiveX orXfound on the AIRSystemand a higher resolutioncamerawillenhancetheimagingcapabilitiesofthesystemForexternalvalidationofHRCellRaftArrayandCLEARAIRSystemperformancewehaveidentifiedtwoinvestigatorsfromourEarlyAdopterProgramwithresearch needs uniquely satisfied by the HRCellRaft and CLEAR AIR SystemOne will isolate single cells basedon subcellular localization of a reporter protein and the second will isolate cells with varying levels of transgeneexpressionwithbothgroupscouplingthesephenotypicobservationstodownstreamsinglecellmolecularanalysisTheHRCellRaftandtheimprovedopticalsubsystemintheCLEARAIRSystemmeetsanurgentlyunmet need for pairing highresolution phenotypic imaging with downstream molecular analysis of single cells