Case study of the automation options and decisions made in implementing a high-throughput cell based screen using the FLIPR？英文文献资料.docVIP

Journal of Automated Methods Management in Chemistry, Vol. 22,No.5 (September–October 2000) pp.139–142 Case study of the automation options and decisions made in implementing a high- throughput cell based screen using the FLIPR TM Derek J. Hook 951 East Rocky Mouth Lane, Draper, UT 84020, USA. Former Director, Biomolecular Screening,NPSPharmaceuticals, 420Chipeta Way, SaltLake City, UT 84108, USA (GPCRs). They are based on the ability to measure intracellular calcium concentration changes as a result of the secondary signalling responsesof these GPCRsto test substances. The response is monitored through ˉuorescent changes of the intracellular dye Fluo-3 as a responsetochangingcalciumconcentration. This case study examines the automation and process change options available to emerging discovery/development stage phar- maceutical companies when considering implementing sophisticated high-throug hput screens. Generally there are both ?nancial and personnel constraints that have tobeaddressed when implementing state-of-the-art screening technology in smaller companies which generally are not as signi?cant as in large pharmaceutical com- Experimental panies. When NPS Pharmaceuticals considered installing a Molecular Devices FLIPR TM for high-throughput cell based Historically, the equipment used was an 8-channel ˉuorometer with integrated liquid handling, capable of readingonecolumn(eightwells) ofa96-wellmicroplate every 5 minutes. This equipment was constructed in- house from commercially available components, and consistedofaHamiltonMPH22008-probeliquidhand- ler (Hamilton Co., Reno, NV), an X±Y stage to move themicroplateacrossthedeckoftheliquidhandler, and an8-lightpipe?breopticbundleunderneaththemicro- plate toprovide lightpathsfor boththe excitationlight andtofeedtheemissionlighttoasinglephotomultiplier tube (PMT) multiplexed to the 8-?bre optic bundle using a rotating scanning mirror. The photomultiplier signal was detected by a signal processi