欢迎来到天天文库
浏览记录
ID:62075066
大小:4.59 MB
页数:54页
时间:2021-04-14
《最新Scanning and Image Processing by Steve Clough[扫描和图像处理由史蒂夫·克劳夫](43)教学讲义ppt.ppt》由会员上传分享,免费在线阅读,更多相关内容在教育资源-天天文库。
1、ScanningandImageProcessingbySteveClough[扫描和图像处理由史蒂夫·克劳夫](43)GSILumonicscDNAmicroarraysusetwodyeswithwellseparatedemissionspectrasuchasCy3andCy5toallowdirectcomparisonsonsingleslideCellsfromconditionACellsfromconditionBmRNALabelDye1LabelDye2RatioofExpressi
2、onofGenesfromTwoSourcescDNAequalhigherinAhigherinBAfterdoubleclickingtheScanArrayicononthedesktop,thefollowingwindowwillappear.ClickontheLaser1andLaser2tabstoturnonthelasers.Afterlasershavewarmedup,thelasertabsturngreen.Clickonthe‘Configure’buttononthelef
3、t.Thefollowingsmall‘Configure’windowwillappear.Clickon‘ScanProtocols’toselectaprotocoltouseforQuickScanning.ThepurposeofQuickScanningistodeterminethecorrectlaserlevelsneededtohavebalancedintensitiesbetweenCy3andCy5channelsandtofindlevelsthatprovideanicesp
4、readofintensitiesacrossthespectrumwithouthavingtoomanysaturatedspots.Note:thescansdonothavetobeperfectlybalancedbetweenthe2dyesasamathematicalcorrectioncalled‘normalizing’or‘smoothing’willbeperformedlater.However,havingthescanlevelsapproximatelybalancedwi
5、llminimizethemagnitudeofthenormalizationcorrectionfactor.InthisexampleweareusingoneoftheVodkinsoybean9karrayssuchas1021/83,1070,or1088.TobegintheQuickScanningprocess,highlightthe‘9KQUICK’protocolandclick‘Change’Thesearedefaultsettings.Verifythatnobodyhasc
6、hangedthemandclick‘Next’.Thequickscanwillonlycoverasmallportionoftheslideasthisreducesthelikelihoodofbleachingthedyesandyouonlyneedasmallsamplingtodetermineifthecolorsarebalanced.Verifyonlythistopsectionisselectedandthenclick‘Next’.Forourscanner,wefoundth
7、atusingadefaultof:Cy3:80PMTand70LaserCy5:70PMTand70Laserworkswellasaninitialstartingpointforscanning.Changevaluesasneededbyhighlightingthedyethatneedschangingandclicking‘Change’EnternewvaluesAndclick‘OK’Highlightthetwodyesandthenclickonthe‘CopyCurrentArea
8、toallFluorophores’tosetthescanareas.Thesearedefaultsettings.Verifythatnobodyhaschangedthemandclick‘Next’.Click‘Run’intheQuickScanwindow.Thesettingsshouldbeastheyappearbelow.Besureresolutionissetat50umorelseyou’remor
此文档下载收益归作者所有