Zeiss AxioObserver D1 User manual

ZeissAxioObserver.D1Microscope
 Room:3164A
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 StartUpProcedure
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1. Removedustcoverfrommicroscope
2. ToggleFluoArcLampswitchON
3. TogglePowerBarswitchON
4. TurnMicroscopeON
‐bypressingpowerbuttonlocatedonleftsideofmicroscope
5. TurnONcomputer
6. Logonto:ObserverUseraccount(nopassword)
7. OpenProgram:AxioVisionRel.4.8
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ShutDownProcedure
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1. Ifusedoil,gentlyremoveoilfromobjectiveusingneatlyfolded
drylenspaper
2. Put10xobjectiveinplace,andlowerobjective
3. CloseAxioVisionprogram
4. Shutdowncomputer.Waituntilcompletelyturnedoff
5. TurnMicroscopeOFF
6. TogglePowerBarswitchOFF
7. FluoArcLamp:
‐Regularworkhours:setintensityto50%oncontrolpad(F2),
andleavelampON
‐Non‐workhours:TurnlampOFF
8. Covermicroscopewithdustcover,butDON’TCOVERTHEHOT
LAMPHOUSING!!
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Microscope Quick Guide
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ZeissAxioObserver.D1MicroscopeGuide
Room:3164A
StartUpProcedure
1. Removedustcoverfrommicroscope
2ToggleFluoArcLampswitchON
3TogglePowerBarswitchON
4TurnMicroscopeON
‐bypressingpowerbuttonlocatedonleftsideofmicroscope
5TurnONcomputer
6Logonto:ObserverUseraccount(nopassword)
7OpenProgram:AxioVisionRel.4.8
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ConfiguringImagingSetup
1. Ifnotalreadyvisible,opentheWorkareawindow
2. FromtheWorkareamenu,double‐clickonMultidimensionalAcquisitiontoexpandlist,andthen
selectMultidimensionalAcquisition.
3. Underthetab,clickLoadbutton.Selecttheappropriateexperimentset‐updependingon
whatchannelsyouwanttoimage(ieAllChannels).
4. Selectthecolorstab
5. Youcanactivate/deactivatechannelsbyright‐mouseclickingonthechannelnumber
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SetSampleonStageandFocus
Onlyuseslidesthathavebeenthoroughlycleanedofmountingmedium!
1. Choosetheappropriateobjectiveonmicroscope
2. Lightlycleanyourslide(onbothsides)withakimwipeandasmallamountof70%ethanol.
3. Ifusinganoillens,addaverysmalldropofimmersionoil.
4. Loadslideonstage.
5. Bringspecimenintofocusandbringyourareaofinteresttothecenterofthefieldofview.
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CapturinganImage
1. Underthetab,selectyourchannelandclicktheMeasurebutton.Alivepreviewwindowappears.
2. Makesurethe‘Lineardisplayoffullrangeofpossiblevalues’buttonisselected.
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3. Adjusttheexposuretimetogetanappropriatesignalintensity.ClickOKbuttononleftside.
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4. Repeatexposuretimemeasurementforeachchannel,however,adjustthefinefocusforonlyone
channeliethemostimportantone
5. Tocaptureyourfinalimage,pressthebuttonlocatedatthebottomoftheCtab.
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Post‐AcquisitionAdjustments
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Brightness/Contrast
1. Temporarilyturnoffthepseudo‐coloringofthedisplayedimagebypressingthebutton.Each
channelisdisplayedinblack/white;youcanviewonechannelatatime.
2. Clickonthepropertiesbuttonfoundbelowtheimage
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3. WhileunderDisplaytabofpropertieswindow,clicktheMin/Maxbutton,orunderyourimage,press
thebutton.
4. Youcanmakefurtheradjustmentsusingthedisplayedhistogram(iemovetopoflineleft,andbottom
oflineright)
‐NOTE:donotadjusttheGamma
5. Repeatforeachchannel
6. Pressthebutton
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AddingaScalebar
1. Toaddascalebar,pressthebutton,orAnnotationsfromthemainmenuandselectScalebar.
2. Dragcursoralongimagetogetdesiredscalebarsize
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SavingFiles
1. Fromthemainmenu,selectFileSaveas
2. SaveyourfilesintheAxioVisionformat(.zvi)intheappropriatedestinationyourlabfolderyour
folder.PleaserefertotheAttentionUserssignspostedonthewall.
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ShutDownProcedure
8Ifusedoil,gentlyremoveoilfromobjectiveusingneatlyfoldeddrylenspaper
9Put10xobjectiveinplace,andlowerobjective
10 CloseAxioVisionprogram.Donotsavechangesmadetotheexperiment!
11 Shutdowncomputer.Waituntilcompletelyturnedoff
12 TurnMicroscopeOFF
13 TogglePowerBarswitchOFF
14 FluoArcLamp:
‐Regularworkhours:setintensityto50%oncontrolpad(F2),andleavelampON
‐Non‐workhours:TurnlampOFF
15 Covermicroscopewithdustcover,butDON’TCOVERTHEHOTLAMPHOUSING!!
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MicroscopeControls
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DisplayScreen
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TransmittedLightControls
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ReflectedLightControls
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LightPath
Locatedonleftsideofmicroscope
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ObjectiveType
ObjectiveNA
ObjectiveContrastMethod
ObjectiveMagnification
CondenserContrastMethod
HALLampVoltage
TLShutter(open)
RLShutter(closed)
Reflector
Locatedonbottomfrontof
microscope
ControlsHALlamp
voltage/intensity
Dia
p
hra
g
mControl
Condensercontrast
method
Condensera
p
erturecontrol
Locatedonbottomright
ofmicroscope
Opens/closesFLshutter
TurnsHALLampon/off
Locatedonbottomright
ofmicroscope
Locatedatbackofright
focusknob
Movesreflectorturret
50%Camera
50%Ocular
100%Camera 100%Ocular

Objectives
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ZeissAxioObserver.D1MicroscopeObjectives
Room:3164A
 123
 A‐Plan10xLDPlan‐NeoFluar20xPlan‐Apochromat
20x
0.25Ph10.4KorrPh2‐M270.8M27
Magnification10x20x20x
NumericalAperture0.250.40.8
WorkingDistance(mm)4.27.9atcoverglass0.750.55
Coverglass(mm)0.170‐1.50.17
ImmersionNoneNoneNone
LongDistance(LD)‐ Yes‐
CorrectionRing(Korr)‐ Yes‐
OpticalSystemICSICSICS
Flatness***********
ColorCorrection***********
UVTransmission********
IRTransmission*********
DIC‐ ‐ (DICII)*****
PhaseContrastPh1Ph2‐
Catalog#441031‐0000‐000421351‐9970‐710420650‐9901‐00
 456
 Plan‐NeoFluar
40xPlan‐Apochromat63xPlan‐NeoFluar5x
 1.3OilPh31.4OilDICM270.15
Magnification40x63x5x
NumericalAperture1.31.40.15
WorkingDistance(mm)0.20.1913.6
Coverglass(mm)0.170.170.17
ImmersionOilOilair
OpticalSystemICSICSICS
Flatness************
ColorCorrection************
UVTransmission***********
IRTransmission**********
DIC(notequipped)**(DICIII)*****‐
HighContrastDIC‐ Yes‐
PhaseContrastPh3‐ ‐
Catalog#000000‐1022‐819420782‐9900‐000440320‐0000
ICS=InfinityColorCorrrectedSystem
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Plan‐NeoFluar100x
1.3OilPh3
100x
1.3
0.2
0.17
Oil
ICS
****
****
****
***
‐
‐
Ph3
000000‐1031‐171
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Filter Sets
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ZeissAxioObserverD1MicroscopeFilters
Room3164A
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1. TransmittedLight
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2. FilterSet34(DAPI/BFP)
Excitation:BP390/22
BeamSplitter:FT420
Emission:BP460/50
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3. FilterSet38(Cy2/GFP)
Excitation:BP470/40
BeamSplitter:FT495
Emission:BP525/50
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4. FilterSet46HE(YFP)
Excitation:BP500/25
BeamSplitter:FT515
Emission:BP535/30
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5. FilterSet45(Cy3.5/mcherry)
Excitation:BP560/40
BeamSplitter:FT585
Emission:BP630/75
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6. FilterSet50(Cy5)
Excitation:BP640/30
BeamSplitter:FT660
Emission:BP690/50
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Filter set
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