Olympus Fluoview FV1000 Bedienungsanleitung

OlympusFV1000Userguide August3,2011

Olympus FV1000 MPE Microscope User Guide

A.SystemStartUp

1. Signintothelogbook.Indicatethelaserlinesyouwilluse;visiblelaserandIRlaser.

2. Turnonmercurylamppowersupplyb. 

3. Turnonmicroscopecontrollerandscanner.

Microscopecontroller‐switchONc

Scanner–switchONandturnKeytoONd.

4. Turnonlasercombinersef.

5. Turnonlaserpowersupplies

Multi‐Arg:SwitchtoONandturnKeytoON.

HeNeh:TurnKeytoON.

6. Turnoncomputeriifitisnoton.

7. Enterusername/passwordtologontoWindowsXP.

8. StarttheFV10‐ASWprogramandenterUserIDand

password.



d

OlympusFV1000Userguide August3,2011

B.OutlineofAcquisitionSetting/ImageAcquisition/ImageViewerWindows

OlympusFV1000Userguide August3,2011



C.ViewingwithTransmittedorEpifluorescenceLight

Beforescanningthesamplewithlaserlight,lookat

thesampleandfindtheregionofinterestfirstwith

eithertransmittedorfluorescentlight.



Clickontransmittedlightbuttonj fortransmitted

lightviewingoronepifluorescencelightbutton1)for

fluorescenceviewing(Beforeclickonthelightbutton

youwillused,clickoffthepressedbuttonfirst).



Turnthefilterwheel1!to#6(DICT)fortransmitted

lightviewing,orto#3(GreenNIBA),4(RedWIG),or

5(DAPI)forfluorescentlightviewing.Openthe

shutter1@.



MAKESURETHESLIDEANDCOVERGLASSARECLEANANDSEALED.Place

theslideonthemicroscopestage.Focusontothesample

withthefocusknob1#(fineadjustment)orfocusbuttons1$

(fastfocusing)ontheremotecontroller(Note:thefocusknobonthemicroscopedoesn’tworkiftheremote

controllerisused).Theextent(fineorcoarse)ofmovementbyfocusknob1#canbeselectedalternativelyby

pressingF/Cbutton1%.

PressEscbutton1^tomovetheobjectivecompletelyawayfromthesampleandpressagaintobringthe

objectivetotheoriginalposition.

Adjustthetransmittedlightlevelwiththelampcontroller1& ontheImageAcquisitioncontrolwindow.

Whenyouarereadyforlaserscanning,turnthefilterwheel1! to#1position(LaserScanninglabel).

Folder/File

Explorer

Datamanager:displays

metadatacontaining

acquisitionparameters.

j

1) 1!

OlympusFV1000Userguide August3,2011

D.ImageAcquisition

Clickoffthelightsourcebuttontoturnoff

thenon‐laserlight.

ClickonDyelistbuttonanddoubleclick

onthefluorescentdyesyouwanttouse(e.g.

FITCandTRITC)fromthelist.ClickApply

buttonb(ifAssignDyeManuallybuttoncis

checked,uncheckbeforepressingApply

button).Itwillactivatethelaserlinesandset

detectionchannelsupaccordingtothedyes

youselected.



ChooseImagesize(default‐512x512)from

Sizepanel d.



Pressbutton(ifnotpressed)to

automaticallyadjustHVandOffsetvalues

accordingtothescanspeedchange.Click

buttontoscanthesample.Itwill

scanfastat2us/pixelandshowlow‐quality

(pixelated)imageinaLiveViewwindow.



Whilescanning,focusontotheregionofinterestwiththeremotefocuscontrollerorbyclickingarrowhead

buttonsinthemicroscopewindow(largearrowheadmovetheobjectivebytheextentsetbyStepSizee,

andthesmallarrowheadsmovestheobjectivebythehalfofthelargearrowheadstepsize).

ClickStopbutton tostopscanning.



SettheLaseroutputlevelf(settheMulti‐Arlaser

to1‐5%andtheHeNelaserto1‐25%,depending

onyoursamplecondition,butnolowerthan1%).



Setthescanspeedg to10~12.5us/pixelandclick

XYrepeatbuttontoscan(youcanscan

imagefasterorslower,dependingonyoursample).



Detector(CHS)ChannelSetting:Adjustthe

brightnessbyHV(andGain)andbackgroundblack

level(Offset)ofindividualchannels.KeeptheHV

valuebelow700(beyond700,thebackground

noisewillshowup).





c

d

g

b

f

h

OlympusFV1000Userguide August3,2011

YoucanoptimizetheimagequalitywithHi‐LoLUT(Look‐UpTable).ClickLUTbuttontobringtheLUT

controlwindowandselectChannelandclickHi‐Lobutton(Ctrl‐H

shortcutkeywilldothesamewithoutopeningLUTwindow).Now

theimageisdisplayedusingthisspecificlook‐uptablewherered

pixelsrepresentintensitybeyondscale(saturated)andbluepixels

represent0pixelvalue.Tomaximizethesignal‐to‐noiseratioofthe

image,adjusttheacquisitionsetting(laserpowerlevel,HV,Offset)

suchthattheimageshowsafewredandbluepixelswhileyouare

scanning.



Oftenexcitation/emissionprofilesoffluorophoresyouareusingmay

beclose,sotherecouldbepossiblebleed‐throughofafluorophore

emissiontoneighboringchannel.Tominimizethis,thedetecting

rangeofeachchannelcanbeadjustedbychangingSpectralSetting.

Clickon,itwillbringSpectralSettingwindow.Changetherange

ofspectrumforeachchannel(CHS1andCHS2)bysliding,widening,

ornarrowingthetabsorarrowheads.



UseAreapanelhtorotateandzoomtheimagingarea(clickon0and1buttonstoreturntotheoriginal

viewingarea).Torotatetheviewingarea,clickonthereddotanddragclock‐orcounter‐clockwise.With

zoomedview,youcanselectthescanningareabymovingtheblue‐linedboxaround.Tozoomaspecificregion

in,clickiconanddrawaboxontheliveviewarea.Itwillzoomintheareaspecifically.



Onceyouaresatisfiedwiththesetting,stop scanning.Setthescanspeedtoslowrates(~10‐12.5us/pixel)

(theslowerthespeedset,thebetterthesignal‐to‐noiseratio,butthemorebleaching).ClickXYbutton

toacquireanimage.Whenacquisitionisdone,a2Dviewwindowwillappear.



E.Z‐SeriesImageAcquisition

Usethismodetoobtainopticalsectionthroughthedepth(z

dimension)ofyoursamplethatcanbeusedfor3Dvisualization.

Clickbuttontoscan.Usethearrowheadsbuttons

orthefinefocusknobontheremotecontrollertofocusinto

differentZ‐axialplanes(largearrowheadbuttonsshiftafullstep

sizeandsmallonesahalfstepsize).Whenyoufindupperlimitof

yoursample,clickEndSetbutton.Bringtheobjectivedownuntil

youfindlowerlimitandclickStartSetbutton.Determinethe

StepSizeandthenumberofSlices,whichcorrelatewitheach

other.Itisrecommendedtosetthestepsizesimilartothe

opticalsectionthicknessoftheobjectiveyouareusing,sothat

thereisnogapbetweentheopticalsectionsuponprojectioninto

3D.Thestepsizecanbefixedbycheckingtheboxb.



c

d

b

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.



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.



F.SequentialScanningMode

Usesequentialscanningtopreventpossible

cross‐talkand/orbleed‐throughbetweenthe

emissionsofthefluorophoresyouused.



ChecktheSequentialboxfanditwillbringup

thesequentialscaninformationwindow.Choose

betweenLineorFramemodeg.



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.



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.



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.



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.



e

OlympusFV1000Userguide August3,2011

G.SavingandExportingImages

1. Clickontheimagewindowtobesaved.



2. Clickon iconorselectFile/SaveorSaveasfrom

menu.



3. ASaveaswindowwillappear.Incasetheimageshould

besavedintheImagefolderofaLog‐inuser,clickon

buttonanditwilldirecttoyourimagefolder.

SelectOlympusImageBinaryFormat*.oib filetype,

typefilename,andclickonSavebutton.(Log‐inuser

anditsassociatefolderwillbecreatedasyoubecome

self‐user).



4. Tosetupyourimagefolderwhereyoucanjumpdirectlyby

clickingMyImageFolderbutton,selectTools/Optionmenu.

ClickGeneral tabandspecifythepathofyourimagefolder

withBrowse.. button.[Uponcreatingauseraccount,each

userwillhaveitsownfolderinDdrive(Donotcreateany

folderinCdrive.).ClickOKbutton.



5. Oibfiletypecontainsallthemetadataincludingallthe

acquisiotnparametersanditcanbeopenedinFV10‐ASW

program.Thelightversionofthisprogramisavailablefor

installinginuser’sowncomputer.Otherwise,theimage

filecanbeexportedasotherfileformatsothatitcanbeopenedandprocessedinotherimagingsoftware.



Toexportimages:

1. SelectFile/ExportorFile/ExportMulti‐Tiffmenu.ClickMy

ImageFolderbuttonifthecurrentfolderisnotyours.



2. WithExportMulti‐Tiffoption,itsavestheimageasa

singletifffilethatcancontainmultipleframes,likez‐or

timelapse‐series.ThisfiletypecanbeopenedinImageJ

software.



ChecktheSavePropertiesAsASCIITextboxtosavethe

metadatathatcontaininformationabouttheimage

acquisitionsetting.Thisdatawillbeusefultofindouttheimagingconditionlaterwithotherprograms.



OlympusFV1000Userguide August3,2011

3. File/Exportcommandbringsawindowwith

differentoptions.

Thiscommandwillexportimagesintoa

folderthatcontainindividualimagesofeach

channel,z‐steps,ortime‐points.



SelectFiletypebforexport.

ROIOverlaycsectionallowssavingthefile

withROIinformation.

OutputFormatd:

RGBColor:thered,green,andbluechannel

willbesavedincolorasindividual*.tiffilesin

afolder.

RGBColor+GrayScaleboxchecked:the

colorchannelswillbesavedingrayscaleas

*.tiffilesinafolder.

MergeChannel:asinglemergedcolorimage

willbesavedasa*.tiffile.



FrameRatee determinetheplayspeedof

themoviewhenthefileisexportedasAVI.

ChecktheAVIboxtocompressthemoviefile.



4. ChecktheSavePropertiesAsASCIITextbuttonifyouexportthemetatdata.ClickSavebutton.



5. TransferyourimagedatatoCDorUSBflashmemory.





c

d e

OlympusFV1000Userguide August3,2011

G.SystemShut‐DownProcedure

Wipeofftheoilfromanyoilobjectivesyouhaveused

duringyoursession.



Loggingoffandtransferringdata.

1. ExittheFV10‐ASWprogram.

2. Makesurethatyoutransferyourimagefileto

CDorUSBflashmemory.Allimagefilesmore

thanamontholdwillberemovedduring

monthlyclean‐up.

3. LogoffWindowsXP.Ifsomeonesignsupforuse

withinanext1hourorso,leavethesystemON.

Ifnot,shutdownthesystemasfollows.



ShuttingOffthesystem.

1. ShutdownthecomputerB.

2. TurnthemercurylampoffC.

3. TurnthemicroscopecontrollerkeytoOFFandturn

offbothscannerandcontrollerI/Oswitchesd.

4. Turnoffthelasercombiners e,FV10‐MCPSUand

FV10‐MP‐LCU.

5. TurningofftheMulti‐Arlaser fbyturningthekey

switchtoOFFfirstandthenturntheI/Opower

switchtoOFF.

6. TurnthekeyofHeNelasergtoOFF.



e

Andere Handbücher für Fluoview FV1000

Andere Olympus Mikroskop Handbücher

Olympus

Olympus CX21 Bedienungsanleitung

Olympus

Olympus POS Bedienungsanleitung

Olympus

Olympus CK40M FRAME Betriebsanleitung

Olympus

Olympus BH2 Series Bedienungsanleitung

Olympus

Olympus BH2 Series Leitfaden

Olympus

Olympus BH2 Series Bedienungsanleitung

Olympus

Olympus U-D7RES Bedienungsanleitung

Olympus

Olympus BX43 Bedienungsanleitung

Olympus

Olympus Fluoview FV1000 Handbuch

Olympus

Olympus CHS Bedienungsanleitung

Olympus

Olympus BX60M Bedienungsanleitung

Olympus

Olympus VM Series Bedienungsanleitung

Olympus

Olympus BX61WI Bedienungsanleitung

Olympus

Olympus SZX16 Installationsanleitung

Olympus

Olympus CHA-P Bedienungsanleitung

Olympus

Olympus DP26 Bedienungsanleitung

Olympus

Olympus BHTU Bedienungsanleitung

Olympus

Olympus U-UVF248A Bedienungsanleitung

Olympus

Olympus U-FWT Bedienungsanleitung

Olympus

Olympus bx40 Bedienungsanleitung

Olympus

Olympus DSX1000 Bedienungsanleitung

Olympus

Olympus CK2 Bedienungsanleitung

Olympus

Olympus IX70 Bedienungsanleitung

Olympus

Olympus CX41 Bedienungsanleitung

Beliebte Mikroskop Handbücher anderer Marken

Thermo Scientific

Thermo Scientific Nicolet iN10 Leitfaden

National

National DC3-163 Bedienungsanleitung

KERN Optics

KERN Optics OPE-1 Bedienungsanleitung

OPTIKA MICROSCOPES

OPTIKA MICROSCOPES B-150D Bedienungsanleitung

Prior

Prior Lux POL Bedienungsanleitung

Opitec

Opitec Smartscoop 115615 Bedienungsanleitung

KERN

KERN OPO-1 Bedienungsanleitung

Topcon

Topcon SP-1P Bedienungsanleitung

Motic

Motic PA43 Bedienungsanleitung

Leica

Leica DVM5000 Benutzerhandbuch

MshOt

MshOt MZ62 Bedienungsanleitung

KERN Optics

KERN Optics OZR-5 Bedienungsanleitung

VWR

VWR VisiScope 384 Series Bedienungsanleitung

Nikon

Nikon ECLIPSE E200 POL Bedienungsanleitung

Leica

Leica DI C800 Handbuch

ThermoFisher Scientific

ThermoFisher Scientific Continuµm Bedienungsanleitung

ThermoFisher Scientific

ThermoFisher Scientific Continuµm Bedienungsanleitung

Leica

Leica FL560 Bedienungsanleitung