BD LSR II and BD LSR II and FACSDiVa FACSDiVa Software Software Dr. Jens Fleischer, Basel Dr. Jens Fleischer, Basel Dr. Norbert Dr. Norbert Leclere Leclere , Berlin , Berlin
BD LSR II and BD LSR II and FACSDiVaFACSDiVa SoftwareSoftware
Dr. Jens Fleischer, BaselDr. Jens Fleischer, BaselDr. Norbert Dr. Norbert LeclereLeclere, Berlin, Berlin
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
OverviewOverviewFluidics
Connectors
On/OffSample PortControlpanel
Electronics Covers
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
TheThe ControlControl PanelPanel
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
EaseEase of of UseUse
• The basic procedures to use the BD LSR II are quite similar to those forFACScan and FACSCalibur
• Controlpanelp with RUN, STDBY and PRIME.
• Flowrates LOW, MED and HI adjustable• Sample injection port with sleeve needle
and tube support arm
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
OpticsOptics
•• TheThe BD LSR II in BD LSR II in youryour lab has lab has threethreelaserslasers::–– 488 nm Coherent Solid488 nm Coherent Solid--State (DPSS)State (DPSS)–– 635 nm JDS 635 nm JDS UniphaseUniphase HeNeHeNe–– 407 nm Coherent 407 nm Coherent VioflameVioflame DiodelaserDiodelaser
•• Using all three excitation line allows the Using all three excitation line allows the user to select the best dyes for the user to select the best dyes for the applicationsapplications
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
OpticsOpticsTheThe BD BD OctagonOctagon technology technology directsdirects thethe collectedcollected light via light via
fibresfibres intointo dedicateddedicated detectiondetection blocksblocks per per laserlaser::
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
““Simple” schematicsSimple” schematics
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
WhatWhat goesgoes wherewhere
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
Default Filters and Default Filters and MirrorsMirrors
Changes in the configuration should be done with care. Use the chartsin the lab to check the setup before starting your measurements!
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
SheathSheath and and WasteWaste
•• TheThe LSR II LSR II isis equippedequipped withwith an an automatedautomated FACSFlowFACSFlow SupplySupply SystemSystem–– 20 L 20 L FACSFlowFACSFlow cubitainercubitainer as as SheathSheath–– 20 L 20 L emptyempty cubitainercubitainer forfor wastewaste–– Level Level sensorssensors withwith audibleaudible alarmalarm–– EasierEasier to to useuse thanthan refillingrefilling and and emptyingemptying
smallsmall tankstanks all 3 all 3 hourshours
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
StartupStartup
•• Check Check SheathSheath cubitainercubitainer, , replacereplace ifif necessarynecessary•• Check Check WasteWaste cubitainercubitainer, , replacereplace ifif necessarynecessary•• First, turn on First, turn on thethe BD BD FACSFlowFACSFlow SupplySupply SystemSystem•• Second, turn on Second, turn on thethe BD LSR IIBD LSR II•• VentVent sheathsheath tubingtubing at at thethe sideside of of thethe instrumentinstrument. Be . Be
carefulcareful notnot to to spillspill PBS PBS overover monitormonitor and and keyboardkeyboard!!•• Take off Take off thethe cleaningcleaning tubetube•• Press PRIME Press PRIME threethree timestimes•• PutPut a a tubetube withwith DI DI ontoonto thethe SIP and SIP and runrun itit forfor 10 10
minutesminutes in HI and RUNin HI and RUN•• System ready for measurementsSystem ready for measurements
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
ShutdownShutdown
•• Run a Run a tubetube withwith FACSCleanFACSClean forfor 10 10 minutesminutes in HIin HI•• PutPut thethe tubetube supportsupport arm to arm to eithereither sideside and and emptyempty thethe remainsremains of of
thethe FACSCleanFACSClean solutionsolution•• ThisThis will will disinfectdisinfect thethe needleneedle and and thethe outerouter sleevesleeve•• Run a Run a tubetube withwith FACSRinseFACSRinse 10 10 minutesminutes in HIin HI•• PutPut thethe tubetube supportsupport arm to arm to eithereither sideside and and emptyempty thethe remainsremains of of
thethe FACSRinseFACSRinse solutionsolution untiluntil appproxappprox. 1 ml . 1 ml isis leftleft..•• ThisThis will will washwash out out deaddead cellscells and and debrisdebris fromfrom thethe needleneedle and and thethe
outerouter sleevesleeve•• PutPut a a tubetube withwith DI DI ontoonto thethe SIP and out SIP and out thethe instrumentinstrument to to
STDBYSTDBY•• Turn Turn thethe BD LSR II and BD LSR II and thethe FACSFlowFACSFlow SupplySupply System offSystem off
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
FACSDiVa SoftwareFACSDiVa Software
•• TheThe newnew digital digital acquisitionacquisition platformplatformintroducesintroduces a a completecomplete newnew software software conceptconcept fromfrom BDBD
•• TheThe software software runsruns on a Windows XP on a Windows XP computercomputer, Macintosh systems , Macintosh systems areare no no longerlonger availableavailable
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
FACSDiVa SoftwareFACSDiVa Software
•• TheThe software software usesuses a a databasedatabase serverserversoftware to manage software to manage thethe flowflow cytometry cytometry datadata..
•• All All datadata, , plotsplots, , gatesgates, , instrumentinstrument settingssettings, and , and statisticsstatistics areare savedsaved insideinside thethe databasedatabase..
•• TheThe useruser cannotcannot specifyspecify anyany pathpath forfor savingsaving, , everythingeverything isis donedone automaticallyautomatically in in thethebackgroundbackground..
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
TheThe mainmain layoutlayout
The BrowserManagement of your
experimental data and settings
InstrumentVoltage settings, Threshold,
Compensation
InspectorAlways shows the properties
of selected objects
WorksheetThe area for plots, histograms,Statistics and everything else
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
„„Floating Floating windowswindows“ (“ (alwaysalways on on toptop))
Acquisition StatusCounters for events per second,
Aborted events,Elapsed time
Acquisition ControlStarts and stops Acquisition and
data recording.
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
LogLog--inin
•• TheThe softwaressoftwares requiresrequires a a useruser loglog--in. Users in. Users cancan onlyonly seesee and and manipulatemanipulate theirtheir ownown datadata. .
•• OnlyOnly thethe Administrator Administrator cancan definedefine newnew usersusersand and theirtheir rightsrights..
•• TheThe Administrator Administrator cancan „Share“ „Share“ datadata withwith otherotherusersusers..
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
LogLog--inin
•• EachEach time time thethe software software isis startedstarted, , thethe loglog--in in menumenu appearsappears::
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
LogLog--inin•• DependingDepending on on thethe loglog--in, in, thethe useruser cancan
onlyonly seesee thethe appropriateappropriate datasetsdatasets::
A user can only seehis own data.
The Administrator cansee everything
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
HowHow to start?to start?
New New experimentsexperiments cancan bebe createdcreated via via thethe associatedassociatedbrowserbrowser iconicon oror via via thethe Menu bar.Menu bar.
New Experiment
Folders can be used to sort data sets. The foldersexist only in the database!
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
Global Global WorksheetsWorksheets
Be Be defaultdefault „Global „Global WorkSheetsWorkSheets“ “ offeroffer thethe possibilitypossibility to „to „rere--useuse“ “ plotsplots likelike in in CellQuestCellQuest. .
WhenWhen pressingpressing thethe „NEXT“ „NEXT“ buttonbutton, , thethe samesame plotsplots areare usedused forfordatadata displaydisplay..
Also Also savedsaved datadata cancan bebe loadedloadedintointo plotsplots..
For different For different viewsviews on on thethe samesamedatasetsdatasets youyou cancan createcreatemultiple global multiple global sheetssheets..
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
A A newnew experimentexperimentA A newnew experimentexperiment containscontains a „a „standardstandard“ “ instrumentinstrument
settingsetting and a and a singlesingle tube in a tube in a soso--calledcalled „„specimenspecimen“. “. WhenWhen selectingselecting "New Experiment" "New Experiment" fromfrom thethe menumenu bar bar
youyou cancan selectselect a "a "TemplateTemplate" " forfor youryour Experiment Experiment ((createdcreated fromfrom youryour olderolder experimentsexperiments).).
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
SpecimensSpecimensTheThe FACSDiVa software FACSDiVa software usesuses „„SpecimensSpecimens“ to “ to associateassociate
a a seriesseries of of tubestubes as a as a patternpattern oror panelpanel. . ExistingExisting SpecimensSpecimens cancan bebe usedused as a as a templatetemplate forfor thethe
nextnext, , similarsimilar tube tube patternpattern..
It is possible to “pre-define” a tube list, but you can always duplicate a specimen after measurement of all associated tubes.
Pre-definition is not required.
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
TheThe firstfirst plotsplotsAt At thethe toptop of of thethe WorksheetWorksheet WindowWindow youyou will find will find thethe "Tool "Tool
Palette"Palette"
Clicking on the “Clicking on the “DotPlotDotPlot” icon activates the creation of ” icon activates the creation of DotPlotsDotPlots. Just click once on the worksheet to create a . Just click once on the worksheet to create a dotplotdotplot of “standardof “standard--size”. Dragging and holding the size”. Dragging and holding the mouse creates a mouse creates a dotplotdotplot with the size of with the size of courcour choice. choice.
In addition do In addition do dotplotsdotplots, the software offers contour/density , the software offers contour/density plots and histograms.plots and histograms.
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
DotPlotsDotPlotsThe DotPlot can be activated byclicking onto the white border area.
The Inspector window shows informations about the activated plot(s):
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
Acquiring DataAcquiring DataDataData acquisitionacquisition isis controlledcontrolled via via thethe „„AcquisitionAcquisition
ControlControl“ “ windowwindow
Data acquisition is started and stopped with the „Acquire“ Button
During acquisition, data saving is initiated by pressing the„Record“ button.
The „Restart“ button is available during acquisition to restartthe acquisition and/or data recording.
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
AcquiringAcquiring datadata
The acquisition control window shows also the tube name, an event counterand the elapsed time since the acquisition has started.
The user also controls which events are counted and stored:
Storage Gate: Which events are recorded and saved
Stopping Gate: Which cells are counted
Events To Record: How many cells should be recorded (in the Stopping Gate)
Events to Display: How many events are shown “on-line” in the acquisition plots
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
TheThe instrumentinstrument controlcontrolTheThe „Instrument“ „Instrument“ windowwindow controls all controls all parametersparameters of of
thethe instrumentinstrument. . ItIt consistsconsists of different of different tabstabs, , whichwhich all all relaterelate to to specificspecific instrumentinstrument functionsfunctions..
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
Instrument Instrument -- ParametersParameters
TheThe Parameter Parameter tabtab showsshowsthethe instrumentinstrumentparametersparameters and and theirtheirsettingssettings. . TheThe useruser cancanchangechange thethe PMT PMT voltagevoltage, , thethelinear/linear/logarithmiclogarithmicsettingssettings, and , and cancanselectselect whetherwhether AreaArea, , HeightHeight and and WidthWidth forforthethe parameterparameter shouldshouldbebe acquiredacquired. . TheThedigital digital electronicselectronics has has no no parameterparameterlimitationlimitation!!
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
Instrument ParametersInstrument Parameters
The „Add“ and „Delete“ buttons allow the user to delete unwantedparameters, thus savingspace for the recordeddata sets.
Note that the parameterscorrespond to fluorochrome names, notchannel numbers. Theassociation betweenchannels, lasers and fluorochromes isexplained later.
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
Instrument Instrument -- ThresholdThreshold
The digital electronicsallows the user to definemultiple thresholds.
Thresholds can be set on any parameter from anylaser.
Threshold can beconnected with logicalOR and logical AND, so that either one of thethreshold conditions isfulfilled or all conditionsmust be met by an event.
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
Instrument Instrument -- CompensationCompensation
The digital electronicsallows the compensation of all parameters againsteach other. Thus, thecompensation starts to getincreasingly complex in multicolor applications.
The software allows„Autocompensation“ ifsingle color stains areavailable, reducing theworkload for instrumentsetup during multicolorexperiments.
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
Instrument Instrument -- RatioRatio
The instrument canautomatically calculate theratio between any twoparameters, even fromdifferent lasers.
Calcium flux:Ratio Fluoro3/FuraRed
Ratio Indo(bound)/Indo(Free)
Absorption ratio for dyesexcitable by different wavelengths
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
Instrument Instrument -- LasersLasers
Users should in mostcases not change thesesettings. All values aresetup during installation of your machine.
Please use this tab only if aBD representative asksyou to do so.
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
Instrument Instrument -- StatusStatus
The Status tab shows anyerrors that the instrumentencounters.
If an error occurs, the color ofthe tab changes to red, signalling that something iswrong.
The Clear button at the lowerright corner deletes thewarning messages.
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
AcquisitionAcquisition StatusStatus
The Acquisition Status window included a counter for events over the threshold(Events/Second) and a sum counter (all eventssince Acquire has been pressed).
The Abort Rate and Abort Count show howmany cells are „lost“ from the analysis becausethey were too close together to be separated as single events.
Processed Events shows how many cells havebeen analysed and is always close to theThreshold Count.
Elapsed Time shows how long you have beenacquiring the data.
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
WorkingWorking WithoutWithout Global Global WorksheetsWorksheets
In order to In order to switchswitch to to thethe „Standard „Standard WorkSheetsWorkSheets“, “, clickclick on on thethe smallsmall greengreentriangletriangle in in thethe upperupper leftleft cornercorner of of thethe tooltool palettepalette..
NowNow youyou cancan createcreate different different plotsplots forfor eacheach tube. In tube. In additionaddition, , eacheach tube tube nownow has has itsits ownown setset of of populationpopulation..
„Standard „Standard WorksheetsWorksheets“ “ areare good good forfor a a longlong list of list of tubestubes withwith different different analysisanalysis requirementsrequirements, , oror ifif youyou wantwant to to putput plotsplots of different of different tubestubesonztoonzto thethe samesame printprint pagepage
HoweverHowever, , bebe carefulcareful whichwhich mode mode isis currentlycurrently activeactive: A : A greengreen tabtab meansmeans"Global "Global WorksheetWorksheet", a ", a greygrey tabtab „Normal „Normal WorksheetWorksheet".".
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
User User PreferencesPreferences• Tube specific Worksheet: Each Tube gets itsown Worksheet
• Start Acquisition on pointer change: Savesone mouseclick on Acquisition
• Show GUID: Unique ID for each tube (useful?)
• Remove Tube-Specific Instrument Settingson Duplicate: Always use the sameInstrument Setting in an Experiment, evenwhen one tube is changed.
•Save Analysis after Recording: Copies all Plots from the Template onto the normal Worksheet after Recording
• Automatically Update Instrument Setting: Use always the latest setup even in "old" experiments
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
User User PreferencesPreferences
• Interval Gate Default Color: HistogramGates per Default have no color to reduce clutter
• Quadrant Gate color: Quadrants arealso not colored to reduce datacluttering
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
User User PreferencesPreferences
•• Default Background Color Default Background Color sincesince versionversion 4.1 4.1 isis „White“ „White“ ((equalequal to to printoutprintout))
•• CanCan bebe setset to to anyany colorcolor bybythethe useruser
•• In In somesome instancesinstances, , especiallyespecially whenwhen updatingupdatingexistingexisting databasesdatabases, , itit cancanoccuroccur thatthat youyou getget blackblackdotsdots on on blackblack backgroundbackground, , alwaysalways openopen thethe Population Population HierarchyHierarchy to check to check thisthis!!
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
User User PreferencesPreferences
•• EachEach useruser cancan definedefine his his ownowndefaultdefault templatestemplates forfor newnewexperimentsexperiments..
•• Default Global Default Global WorksheetWorksheet: BD : BD recommendsrecommends to to useuse thethe global global worksheetworksheet as as standardstandard methodmethod to to reducereduce screenscreen clutterclutter..
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
ExportingExporting TemplatesTemplates
After choosing "Export" the user can select the folder in a drop-down box. If you type in a new name, the folder isgenerated.
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
New New InspectorInspector viewview forfor recordedrecordedtubestubes
•• TheThe newnew inspectorinspector viewview forforrecordedrecorded tubestubes showsshows thetheinstrumentinstrument settingssettings in a different in a different way way comparedcompared to to thethe „live „live settingssettings“ “ shownshown beforebefore
•• ItIt isis notnot possiblepossible to to changechange thethevolatevolate settingssettings oror to to deletedelete / / addaddparametersparameters AFTER AFTER acquisitionacquisition
•• ItIt isis possiblepossible to to chnagechnagecompensationcompensation offlineoffline
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
New New scalingscalingIn In thethe digital digital electronicselectronics therethere areare no no errorserrors in in thethe determinationdetermination of of
thethe logarithmiclogarithmic valuevalue forfor lowlow intensitiesintensities. . IndeedIndeed, , itit alwaysalways lookslooksup up thethe mathematicallymathematically correctcorrect valuevalue..
HoweverHowever, , thethe logarithmiclogarithmic scalescale cancan onlyonly displaydisplay numbersnumbers fromfrom 26 26 to 260.000 in to 260.000 in thethe 44--decade decade displaydisplay ((whichwhich isis recommendedrecommended).).
In In thethe 55--decade decade displaydisplay itit isis 2.6 to 260.000 2.6 to 260.000 channelschannels. .
TheThe electronicselectronics actuallyactually calculatescalculates intensitiesintensities fromfrom ––2600 to 2600 to 260.000. 260.000. EverthingEverthing belowbelow 26 in 26 in thethe 44--decade decade scalingscaling thereforetherefore isisan "an "onon--axisaxis" " eventevent, as , as therethere isis no no logarithmiclogarithmic valuevalue to to plotplot..
TheThe "way out": "way out": UseUse a different a different axisaxis scalingscaling. . TheThe "bi"bi--exponential" exponential" sclaingsclaing isis linear at linear at thethe beginningbeginning and and convertsconverts to to logarithmiclogarithmic at a at a certaincertain point.point.
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
ExampleExample 1 1 –– Normal Normal loglog--scalescale
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
ExampleExample 1 1 –– BiBi--exponentialexponential
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
ExampleExample 2 2 –– Normal Normal loglog--scalescale
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
ExampleExample 2 2 –– BiBi--exponentialexponential
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
ExampleExample 3 3 –– Normal Normal loglog--scalescale
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
ExampleExample 3 3 –– BiBi--exponentialexponential
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
BiBi--ExponentialExponential
BiBi--exponential exponential scalingscalingshowsshows also also nicelynicelythethe effecteffect of of compensationcompensation..
TheThe upperupper panelpanel isisoverover--compensatedcompensated, , thethe effecteffect isis clearlyclearlyvisiblevisible in in thethe bibi--expoentialexpoential displaydisplay..
CorrectCorrect CompensationCompensationcancan bebe nicelynicely seenseenin in thethe lowerlower panelpanel..
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
BiBi--Exponential Exponential –– WhereWhere to to activateactivate??
The bi-exponential scaling is activated for each plot in theInspector. Thus, it is possible to have bi-exponential and normal log scale on the same page.
Population borders drawn in bi-exponential displays cannot be displayed in "normal" plots, but are always calculated on bi-exponential scaling.
The same holds true for the borders of population from "normal" log scales. They are not drawn in bi-exponential plots, but calculated based on log scale.
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
DensityDensity PlotsPlots
Densities and Contour Plots canbe generated with the same icon
Final formatting is done via the„Inspector“ Window.
Note that density plots can also be calculated on bi-exponential scaling, showing the populationdistribution.
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
CopyCopy and Paste...and Paste...
TheThe FACSDiVa Software FACSDiVa Software nownow supportssupports CopyCopy and Paste and Paste to to generategenerate PowerPoint PowerPoint PresentationsPresentations oror WordWord--DocumentsDocuments ((oror anyany otherother formatformat) ) withwith flowflow cytometry cytometry datadata..
Alternatively, Plots can be exported as graphics files. Alternatively, Plots can be exported as graphics files. However, only JPGHowever, only JPG--format is supported, leading to format is supported, leading to loss of quality if resized after import to PowerPoint.loss of quality if resized after import to PowerPoint.
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
Hinged QuadrantsHinged Quadrants
In many cases rectangular quadrantsare not the correct solution.
The FACSDiVa Software now offers "hinged" quadrants, which can be moved individually to gate the cells of interest without the need to create four different polygon regions.
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
Hinged QuadrantsHinged Quadrants
In many cases rectangular quadrantsare not the correct solution.
The FACSDiVa Software now offers "hinged" quadrants, which can be moved individually to gate the cells of interest without the neeed to create four different polygon regions.
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
OffsetOffset QuadrantsQuadrants
•• SometimesSometimes itit maymay bebeusefuluseful to to offsetoffset oneone sidesideof of thethe quadrantsquadrants to to adjustadjust clearlyclearlyoverlappingoverlapping populationspopulations..
•• YouYou cancan eithereither offesetoffesetquadrantsquadrants oror useuse thethehingedhinged versionversion. . ItIt isis notnotpossiblepossible to to useuse bothbothversionsversions simultaneouslysimultaneously!!
Dr. Jens FleischerBDB European Technology Center
Basel, Switzerland LSR II Basic Course Dresden January 2006
Final Final recommendationrecommendation::
Just Do It!