?url_ver=Z39.88-2004&rft_val_fmt=info%3Aofi%2Ffmt%3Akev%3Amtx%3Adc&rft.relation=http%3A%2F%2Fd-scholarship-dev.library.pitt.edu%2F7903%2F&rft.title=ONLINE+DETERMINATION+OF+EXTRACELLULAR+GLUTATHIONE+IN+ORGANOTYPIC+HIPPOCAMPAL+SLICE+CULTURES+WITH+A+MCIROFLUIDIC+DEVICE+AND+CONFOCAL+LASER-INDUCED+FLUORESCENCE+DETECTION&rft.creator=Wu%2C+Juanfang&rft.description=GSH+is+an+essential+cellular+antioxidant+in+the+brain+cells+due+to+its+active+sulfhydryl+group.+Measuring+the+concentration%2C+distribution+and+metabolic+pathways+of+GSH+and+related+compounds+provides+a+lot+of+useful+information+for+studying+oxidative+insult+in+vivo.+Microfluidic+chips+are+ideal+platforms+for+biological+samples%2C+because+they+are+capable+of+analyzing+very+small+amount+of+analyte+in+a+relative+short+time.+The+coupling+of+microfluidic+separation+and+laser-induced+fluorescence+provides+an+effective+way+for+analyzing+GSH+and+related+compounds.+A+confocal+LIF+detection+system+has+been+developed+in+our+laboratory.+The+key+part+in+building+this+system+is+the+selection+of+an+optimized+set+of+optical+components%2C+which+mainly+includes+a+laser%2C+an+objective%2C+a+front+surface+mirror%2C+lens%2C+filter+sets+and+a+detector.+The+parameters+of+these+optical+components+were+first+optimized+according+to+the+preliminary+scheme+of+the+system.+Meanwhile%2C+an+opto-mechanical+assembly+was+designed+and+made+for+arranging+these+components+into+a+whole+system.+A+controlling+program+in+LabVIEW+was+also+developed%2C+which+incorporates+several+major+functions%2C+such+as+data+collection+and+pretreatment%2C+voltage+and+current+monitoring%2C+high+voltage+power+supply+(HVPS)+manipulation%2C+sampling+control%2C+into+one+program+and+offers+an+easy+access+for+user+to+have+a+convenient+control+of+the+whole+system.+The+procedures+of+beam+alignment+for+our+detection+system+are+discussed+in+detail+in+the+system+testing+section.+The+profiles+of+the+focused+laser+beam+at+three+different+positions+were+measured+by+using+a+simple+setup%2C+the+results+of+which+were+consistent+with+those+from+the+theoretical+calculation.+A+series+of+experiments+were+designed+to+identify+the+dominant+noise+sources+in+the+system+for+a+better+signal+to+noise+ratio+(SNR).+Noise+reduction+was+finally+obtained+by+applying+averaging+and+low+pass+filter.+Experiments+with+real+samples+showed+that+the+linear+range+for+GSH+labeled+with+Thioglo+1+is+between+0-49.7+%C2%B5M+and+the+limit+of+detection+(LOD)+is+around+1.45+nM+for+our+system.+Our+future+work+will+be+focused+on+connecting+electroosmotic+sampling+with+separation+on+microfluidic+device+to+analyze+the+extracellular+GSH+in+organotypic+hipocampal+slice+cultures.&rft.date=2010-09-20&rft.type=University+of+Pittsburgh+ETD&rft.type=PeerReviewed&rft.format=application%2Fpdf&rft.language=en&rft.identifier=http%3A%2F%2Fd-scholarship-dev.library.pitt.edu%2F7903%2F1%2FJuanfangWu090908.pdf&rft.identifier=++Wu%2C+Juanfang++(2010)+ONLINE+DETERMINATION+OF+EXTRACELLULAR+GLUTATHIONE+IN+ORGANOTYPIC+HIPPOCAMPAL+SLICE+CULTURES+WITH+A+MCIROFLUIDIC+DEVICE+AND+CONFOCAL+LASER-INDUCED+FLUORESCENCE+DETECTION.++Master's+Thesis%2C+University+of+Pittsburgh.++++(Unpublished)++