Beckman流式细胞仪-GALLIOS使用说明书

Gallios TM Quick ReferencePurpose:The purpose of this Quick Reference is to provide a simple step by stepoutline of the information needed to perform various tasks on the system.We begin with basic tasks like starting up or shutting down the system andthen proceed to more involved software tasks. It is organized according tothe modules in the Gallios Flow Cytometer Training Modules. MajorHeadings (in blue) are the major groups of tasks. These headings may befurther subdivided into subsections of the major task and appear in alogical sequence. For example, the Major task of creating a protocol issubdivided into the subsections: Select Parameters, Create Plots,Instrument Settings, Create Regions, Gate, Analyze, and Save Protocol. Alist of the major tasks appears below in the Table of Contents.Table of ContentsStartups P. 3Shutdown P. 4Creating a Protocol P. 5Create a FlowPAGE P. 7AutoSetup P. 8Acquisition Manager (Panels) P. 9Database P. 11Offline Analysis P. 15System S etup P. 19StartupsCheck external Sheath tank, fill the Cleaning agent tankand Empty the Waste tankPower the Computer and Monitor OnEnter a Windows password if any and select OKDouble click icon.Select User nameEnter User password and select NextSelect Connect (to run samples) or W ork Offline .Check gages System Pressure 30 Psi, Vac 17 in (Cytometeronly)Commonly Used Toolbars:Save LMD Print Copy Undo PRISM Histogram Rectangle Elliptical PRISM Multilinear Gates Color Publish File Plot Plot Divider 2 region Blend Open LMD Save Cut Paste Redo Dot Density Polygon Quadrant Contour Linear PRISM ColorMenusWorkspaceToolbarsAcquisition ManagerResourceExplorer Plot TabsFocusobjectWorklist file tab Panelfile tabProtocols file tab Listmode file tab CurrentWorkspaceprotocolHelp FileToolbarCalculateStatsFile Protocol Plot Plot region Divider 1 Precedence ZoomStart Pause & Rotate Abort Shutdown Clenz Manual Mode Cytometer Control Restart Pause Stop Flow Rate Prime Single TubeFixed ModeOpen Worklist Save Panel Add Tube Customize Worklist AutoschedulerNew Worklist Save Worklist Delete Current Add Panel Print Open ExternalCytometerToolbarTasks Worklist OrderAcquisitionManagerToolbar ShutdownPlace 2 ml of 10 % bleach solution in one tubePlace 2 ml of de-ionized water in each of 3 tubesSelect on the toolbar to clear the Acq. Mgr.Select the tab on the Resource ExplorerSelect the Common FolderSelect the c leanse.pnl and drag and drop it to the Acq.Mgr.Enter a carousel numberSelect buttonSelect Yes if you wish to save the previous protocol or N oif notWhen the panel is finished, remove all tubesSelect Standby modeWhen finished select in the upper right to exit thesoftwareSelect NoSelect to turn off the cytometerSelect Start >> Shutdown >> OK to shutdown thecomputerCreating a ProtocolSelect File >> New >> New Protocol to clear theworkspace of all but one dotplot and single elliptical gateregion.Select Yes to save old protocol or N o if you do not wish tosave itSelect Parameters: ? Select Cytometer Control on the toolbarsSelect Acquisition Setup tabSet Limits and Max events to stop accumulation laterSelect buttonDeselect unwanted parametersSelect any desired parametersOptional: Set Ratio, Time and Derived ParametersClose shutters of unused lasersSelect FS Collection angle and SS desired ND filterSelect OKSelect Yes, enter a name and select Save, Histogram PlotCreate Plots: ? Dot Plots (2 parameter) – Select, Density plot – Select(Single parameter)– SelectSelect Parameter(s) and then OKRepeat for each plotSelect each plot Focus (top of the plot) in turn and drag theplot to the desired location on the WorkspacePress C TRL + T, select plot size, and then OK to arrangeplotsInstrument Settings: ? Select Single Tube Mode on the toolbarPlace a sample for the application in position 10 on thecarouselSelect RunSelect Cytometer Control and then the Acq. Setup tabSelect Setup Mode and QuickSETChoose discriminator parameter and adjust Discriminator toexclude debris and noise but pass cellsAdjust the slider bars (top and side of each plot) beginningwith those to be used as gates for a correct patternSelect Acq. Setup tabIf needed, select QuickCOMP and adjust thecompensation using a proper sample until correct patternsare seenDeselect S etup Mode, QuickSET, and QuickCOMPAllow the system to collect the desired amount of dataSelect to stop or allow the system to reach a stop value Create Regions ? Select each plot in turn and add regions:Dot Plots (2 Parameter Plots only):Polygon - selectMove the cursor to starting point on plot and left clickUse a click move technique to create all the sidesback to the starting pointRectangle– selectMove the cursor to the starting corner and left click Move the cursor to the opposite corner and leftClick to complete the rectangleQuadrant – SelectMove the cursor to plot and left clickMove the region to anchor position and left clickAutgogate– Select ellipse or contourMove the cursor to desired population and left clickHistogram Plots:Linear – SelectMove the cursor to starting position and left clickMove the cursor to ending position and left clickMultilinear– SelectMove the cursor to first position and left clickMove the cursor to end of first region and left clickMove the cursor to end of second region and left click Continue until all regions have been c reatedMove cursor to and left click to finishGate ? Move the cursor to lower right corner of plot to be gatedWhen appears, left click and select GateIf you wish to apply the gate to all plots select that option(Note: later ungate t he plot containing the gate region) Optional: Stop Count ? Select Stop and Save tab and enter countSelect OKAnalyze ? Select A nalysis >> Select ResultsDeselect all unwanted statistics and select all desiredstatistics and Select OKSave Protocol ? Select F ile >> Save Protocol or SelectCreate a FlowPAGESelect Insert >> Blank Flow PageMove the new page to a convenient area of the screenAdd Plots ? Move the cursor to the center of a plotLeft click and drag and drop it onto the FlowPAGERepeat for each plotArrange Plots ? Select the Maximize Window icon in the upper rightcorner of the FlowPAGE to expand the page to full screenMove each plot and move into positionORCreate a large rectangle completely encircling two plotsand statsSelect the appropriate alignment tool (top, bottom,left or right)Add Text ? Select on the FlowPAGE toolbarMove the cursor to the location on the page and left clickType in desired text and select OKReshape the text box by grabbing a handle and pullingoutwardRight click in the box and select Properties to change font,fill, etc.Left click on another part of the page to finishAdd Images ? Select on the FlowPAGE toolbarLocate the directory where the image is storedSelect the desired image and then OpenMove the cursor to the desired location on the page and leftclickUse the handles to resize the imageLeft click on another part of the page to finishAdd Lines or Boxes ? Select orMove to the starting pointHold the left button and drag to the ending pointSave FlowPAGE ? Select F lowPAGE >> Save FlowPAGE AsEnter a name and select SaveAutoSetupNote: To enter a new application, you will need a base protocoland target channels for Flow-Set ProNew Application ? Select T ools >> AutoSetup Application DefinitionSelect Create a new application definitionSelect Current user only or All users as desiredSelect NextBrowse to find the base protocol that represents theapplicationSelect the base protocol and then OpenSelect NextSelect Filter Block and then N extOptional: select dyesSelect NextEnter the correct Flow-Set Pro target channelsSelect NextEnter an application name and select NextSelect Create a verification protocol if you wish the systemto generate a verification protocolOrSelect Add additional verifier, locate the protocol, andselect O pen to add a verification protocolSelect NextThe system summarizes your entriesSelect Print if you wish and then FinishThe system now creates all protocols and FlowPAGESneeded for AutostandardizationSchedule an ? Select T ools >> AutoSetup SchedulerApplication ? Select desired applicationOrHold the CTRL key down and select multiple applicationsEnter a carousel number and select ScheduleSelect Run if you wish to run immediatelyOrSelect Close to prep tubes, place them on the system, orenter a calibration factor in the Acquisition ManagerSelect on the toolbar to run the samplesAcquisition Manager (Panels)If it is not visible, select View (or right click)Select Acquisition ManagerSelect ColumnsSelect View >> Customize Worklist ColumnsOKSelect the desired columns to appear and thenEdit Column Names ? Select V iew >> Customize Worklist ColumnsLeft click on the column name until the name can bechangedType in the new name and press EnterWhen finished, select O KCreate a Panel ? Select to clear the Acquisition ManagerSelect protocol tab on the Resource ExplorerDrag and drop the desired first protocol to the AcquisitionManagerSelect the on the toolbar to add the desired number oftubes to the panelIf you wish to change the protocol in any of the tubes,select the Open Protocol in the Protocol column forthat tubeSelect the desired protocol and then OpenRepeat as necessary for each tubeUnder Regions column, toggle the appropriate icon:to use the regions in that protocol, to carry the regionto the other tubes, or to import regions from anotherprotocolUnder the Cytosettings column, toggle the appropriate icon to:to use the cytosettings in the protocol, to pass thecytosettings to the next protocol, or to import cytosettingsfrom another protocolSelect OpenEdit the Parameter names for each tube to reflect the actualmeasurements (example: CD3, CD4, etc.)To Delete a Protocol ? Select the protocol line by its number and press DeleteTo Move a Protocol ? Select the protocol and drag and drop to the new positionSave Panel ? Right click on Panel folder or panel area inAcq. Mgr.Enter a panel nameIf you desire to create a template in the database forReporting results, select Export results of panel toReport Generator on the Panel Save DialogSelect S aveCreate a Worklist ? Select Clear Worklist to clear the Acq. Mgr.then:To Add Panels ? Select Panels tab on the Resource ExplorerLocate the desired panel and drag and drop it to theAcquisition ManagerRepeat as desiredTo Add Protocols ? Select Protocols tab on the Resource ExplorerLocate the desired protocol and drag and drop it tothe Acquisition ManagerRepeat as desiredEditing Parameter ? Select the desired field (e.g. Under P1, P2, etc)Names ? Type in the desired information (e.g. Sample ID,Parameter name, etc.)Save Worklist ? Select Save Worklist on the toolbarsEnter a worklist name and select SaveTo Run ? Select to clear Acquisition ManagerSelect Worklist tab on Resource ExplorerLocate the desired worklist and drag and drop it tothe Acquisition ManagerEdit the boxes as desiredTo add Cal Factor, click on in the Cal Factorcolumn of the Acquisition ManagerEnter absolute count calibration value, verifyEnabled has been selected, and selectOKEnter a carousel number in tube one positionSelect RunDataBaseQC Product EditorEditing a Product ? From the Report Generator toolbar selectProduct EditorSelect the desired product you wish to editSelect the desired information field to be editedMake the changeRepeat the process for each fieldWhen finished, select S ave Prod then select Y es toconfirm update to the lot numberSelect O KEntering a New ? From the Report Generator toolbar selectProduct ? Product EditorSelect the N ew Product buttonSelect the first information fieldEnter the informationRepeat the process for each fieldWhen finished, select Save Prod then select Y es toconfirm update to the lot numberSelect OKQuality Control Report Sample data is sent to the database when samples are run and the protocol region properties are set correctly Setting Region ? Drag and drop a protocol to the workspaceProperties ? Save the previous protocol or not as desiredSelect a region for statistics to be sent to thedatabaseRight click the mouse and select Region PropertiesSelect Region statistics to be exported for QualityControlSelect a QC Product and select OKRepeat for each region to be usedSave the protocolActivating QC ? Select on the Report GeneratorTemplate toolbar(one time procedure) ? Change instrument to DefaultSelect T emplate >> Select TemplateSelect desired template and then OpenSelect T emplate >> Save Template to save thetemplate with the system serial numberRepeat process for each templateSelect a Template ? Select on the Report Generator toolbarSelect T emplate >> Select TemplateSelect desired template and then OpenSelect a point ? Left click on the point (a red line appears)Creating a New ? First, enter the new QC product (earlier in database) QC Template and ensure the protocol has regions selected for QCexport with a QC productRun the protocol at least onceSelect on the Report Generator toolbarSelect T emplate >> Clear TemplateSelect N ew Plot buttonClick on the header portion of the plotSelect user label and select a label from the list orenter the desired labelSelect the QC protocol from the listSelect the appropriate QC productSelect the lot numberSelect a parameter or regionSelect X or Y axisSelect a statisticWhen finished, select O KRepeat process for each new plotWhen finished, select T emplate >> Save AsEnter a name and select SavePanel TemplateAccessing a template ? Select on the Report Generator toolbarSelect F ile >> Open and locate the desired templateSelect the template and then OpenCreating a Template ? First create a panel and then save the panel with theExport results of panel to report Generator selectedSelect Panel Template on the ReportGenerator toolbarSelect F ile >> Open and locate the new blanktemplateSelect the template and then OpenSelect the desired global options on the Local tabSelect each field in turn and enter the desiredinformationSelect U pdate to enter the information and clear thefieldsOrSelect U pdate & Retain to enter the informationonlyRepeat for each lineSelect Data Plots tab to add plotsWhen finished, select F ile >> SavePanel Report ? Select Panel Report on the Report Generator toolbarSelect S earch button to list reportsHighlight desired report and then click on S elect Database Entry ? Select on the Report Generator toolbarSelect NewEnter information in each desired field and selectSaveMaintenance Log ? Select , double click on item to logService Log ? Select , select field and enter informationOffline AnalysisRetrieve with Runtime ? Select Listmode tab on the Resource Explorer Protocol (protocol used ? Locate the desired listmode file and drag and drop To collect the data) the file to the WorkspaceIf asked do you wish to clear acquisition data,answer Y esSave previous protocol as desiredThe protocol appears and the data shortly after Retrieve with a Different ? Select Protocols tab on the Resource Explorer Protocol ? Locate the desired protocol drag and drop it theWorkspaceIf asked do you wish to clear acquisition data,answer Y esSave previous protocol as desiredSelect Listmode tab on the Resource ExplorerLocate the desired listmodeDrag and drop it to any one of the plots on theWorkspaceThe protocol will populate with the dataRetrieve Multiple Listmode ? Select Tools >> Listmode PlaybackFiles with Runtime ProtocolsRight click on Add Protocol tool in thewindowSelect New WorklistSelect Add Protocol tool once for eachaddition protocolUnder the LMD File column, select Open FileiconSelect the desired listmode file and then OpenRepeat for all listmode filesSelect the Options tab and other desired optionsSelect the Single File Mode tabSelect the Run tool in the windowThe files are displayedFiles with Different Same as previous action but with one addition:ProtocolsSelect the Open Protocol icon under theProtocol columnSelect the desired protocol and then OpenRepeat for each protocolBatch Automator ? Select Tools >> AutoMATOR SetupTo remove old selected files from the list: selectthe files and then the Remove Files buttonSelect A dd FilesSelect the desired files and then OpenTo change the order: Select a file and drag and dropit to a new positionSelect the desired process option next to the files(pass,pause 12 seconds, stop and wait torestart)Select the desired output optionTo process: Select Run on the AutomatortoolbarListmode Playback Tool ? Select Tools >> Listmode PlaybackSelect either the Add Protocol or AddPanelSelect Open for the first listmode fileSelect the listmode file and then OpenRepeat for each listmode fileIf desired, select the Open for each protocol inturnSelect the desired protocol and then OpenSelect Options tab and desired optionsSelect Single File Mode tabSelect tool in the windowThe files are displayedPublishingSet Preferences ? Press C TRL + W and select Publish tabSelect the desired publishing option Publish to TextFile or Publish Data to MS Excel or bothSelect desired data formatSelect desired bitmap sizeSelect desired File References optionSelect whether or not to switch to Excel on publishSelect O KOutput ? Select Tools >> Publish Results NowEnter name and select SaveSystem opens MS Excel and displays data Assigning Color to Dot Plots ? Press C TRL + W and select Gating tab Color Precedence ? Select Automatic Color PrecedenceSelect O KSelect Color Precedence Setup on the toolbarsEdit colors as desiredMove regions using a drag drop techniqueSelect O KColor will be assigned to dots based on the colorlocated highest on Precedence list (top position) Color Blend ? Select C olor Blend ModeSelect OKSelect Color Blend SetupSelect 3 different regions to blendSelect O KColor will be assigned based on the number of theregions satisfied by the eventCopy and PasteCopy Image ? Select a plotRight click and select Copy Plot Image Paste Image ? Open the desired program (e.g. PowerPoint, Word)Select desired areaSelect E dit >> PasteCopy Data ? Select plotRight click and select Copy Plot Data Paste Data ? Open the desired program (e.g. MS Excel)Select desired areaSelect E dit >> PasteFreeze Single Histogram ? Run a sample or retrieve a listmode file Plots ? Choose a histogram plot to apply freeze frameMove the cursor to the lower right corner of the plot Until appearsLeft click and select the Histogram tabSelect Freeze Frame and then OKRun another sample or retrieve another file, anoutline of the previous file appears on top of thenew fileYou can repeat the process to achieve 2 freezeframes on top of a new fileBaseline Offset ? Select F ile >> Workspace PreferencesSelect Plot Display tabSelect B aseline Offset OnSystem adds a randomized positive Gaussian offsetto thelow channels bringing the negative populationout from channel zero on log displaysListmode Compensation ? Select Listmode tab on the Resource ExplorerSelect desired listmode fileDrag and drop it to the workspaceSelect A nalysis >> LMD Quick COMPAdjust the slider bars to change the compensationThe data responds accordinglySave recompensated file (file extension will beCMP)System SetupAdministration Functions ? Select a the user with administration privilegesEnter the user passwordSelect A dmin buttonSelect the user, enter both the user andadministration passwords and select OK Add a User ? Select Add UserEnter user name, first name, last name and titleSelect desired privilegesSelect Change and enter a user passwordSelect O KOptional: Select Paths tab and change user pathsSelect O KGroup Users ? Select W orkgroupsSelect E dit and then AddRename your workgroup as desiredSelect O KSelect all users to be included in the workgroupSelect O KTrack User Usage ? Select E xport LogEnter range of dates and select NextSelect users to be included in the billing fileSelect N extSelect to send to clipboard or file and selectDelimiterSelect F inish and then C loseUnlock a User ? Select userSelect M odify UserSelect U nlock UserArchive ? Select Archive DataSelect Archive locationSelect datesSelect data to archiveSelect A rchive and then O KSelect C loseWorkspace Preferences ? Select F ile >> Workspace Preferences File Naming ? Select LMD Filename tabSelect items to be part of the filenameSelect O KPlot Display ? Select Plot Display tabSelect desired optionsSelect O KGating ? Select Gating tabSelect desired optionsSelect O KAcquisition ? Select Acquisition Options tabSelect desired output optionsSelect O KUser Info ? Select User Info tabEnter desired user informationSelect O KPublish ? Select Publish tabSelect desired publishing format optionsSelect O K。