Olympus FV1000 - Sequential Scanning Mode for Cross-Talk Prevention; Line vs. Frame Sequential Scanning

OlympusFV1000Userguide August3,2011
ClickStop button.AdjusttheScanSpeedifneeded,clickDepthcbutton(“Z”willbeappearontheXY
buttontobecomeXYZ),andthenclickXYZdbutton.
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Whenacquisitionisdone,AppendNext/SeriesDone
e buttonwillappearovertheStopbutton.Click
AppendNextbuttontoaddadditionalsectionsatnext
step(enterthenumberofseriesyouwanttoadd)or
clickSeriesDonetofinishtheacquisition.Savetheimage.
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F.SequentialScanningMode
Usesequentialscanningtopreventpossible
cross‐talkand/orbleed‐throughbetweenthe
emissionsofthefluorophoresyouused.
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ChecktheSequentialboxfanditwillbringup
thesequentialscaninformationwindow.Choose
betweenLineorFramemodeg.
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Line:byline‐by‐line,itscansonechannelwith
onlyaspecificlaserlineanddetectoronandthen
sequentiallyscansthenextchannelwithonlya
anotherlaserlineanddetectoron.
Frame:itscanssequentiallyframe‐by‐frame;it
finishesscanningonechannelfirstandthenscans
thenextchannel.
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Ifthereisnobleed‐through
betweentheemissionsofsome
fluorophoresyouareusing,you
cangroupthemtogethertoscan
themsimultaneouslyasagroup.Simplyclickonthefluorophorenameonthelistanddragtotheothergroup.
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Note:itwillbegoodtosetuptheacquisitionconditionforeachchannelbeforeusingtheSequentialmode.
Todoso,firstinsimultaneousmode(uncheckedSequentialbox),settheacquisitionconditionbyturningon
onlythespecificlaserforthespecificchannel.
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ScanwithFocusx2orXYRepeatbuttontosee
tocheckifthereisanybleed‐through.Ifnot,stopscanning,
andsettheScanspeedataslowerrateandclickXY(Z)button toacquiretheimage.
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