Mitsubishi P91W Manuale delle Istruzioni

ChemiDoc™ MP System with Image Lab™ Software Instruction Manual

ChemiDoc MP with Image Lab Software| ContentsxXcitaBlue™ Conversion Screen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 112Gel Alignmen

ChemiDoc MP with Image Lab Software | Analyzing Images90ALIGNMENTAlign volumes by using the appropriate alignment button. To select several volumes,

ChemiDoc MP with Image Lab Software917 Generating ReportsAfter viewing results, you can generate a report that displays the analyzed gel images, all o

ChemiDoc MP with Image Lab Software | Generating Reports92Use the following dialog boxes to remove information from reports. Doing so does not delete

Report93Image Info• Acquisition Information• Analysis Settings• Image Information•NotesLane and Band Table TabClear any of the following settings to e

ChemiDoc MP with Image Lab Software | Generating Reports94Vo l u me Ta b le TabInclude Volume Table—clear to exclude this information from your report

ChemiDoc MP with Image Lab Software 958 Exporting ResultsThe most convenient way to archive complete information about experiments is to produce repor

ChemiDoc MP with Image Lab Software | Exporting Results96The available export options are described in the following sections.Exporting Gel Images fo

Exporting Gel Images97Note that you can select the entire image or the current view, select the resolution or specify a custom resolution, specify the

ChemiDoc MP with Image Lab Software | Exporting Results98Exporting Gel Images for AnalysisWhen you select File > Export > Export for Analysis,

Exporting Gel Images99Analysis Table ExportYou can export table analysis data from the File menu or by using the export buttons at the top of the Anal

ChemiDoc MP with Image Lab Software11 IntroductionThe ChemiDoc™ MP imaging system offers exceptional application flexibility, high performance, and ea

ChemiDoc MP with Image Lab Software | Exporting Results100

ChemiDoc MP with Image Lab Software 1019 MaintenanceThis chapter includes instructions for maintaining the universal hood in proper working condition

ChemiDoc MP with Image Lab Software | Maintenance1024. Remove the cover with the UV glass by sliding it forward, then lifting up.5. Place it on a non

Fuse Replacement103Fuse ReplacementAlways unplug the instrument before changing or checking the fuses.This unit is protected by 2 fuses (5 x 20 mm, 2

ChemiDoc MP with Image Lab Software | Maintenance104

ChemiDoc MP System with Image Lab Software105Appendix A. Using the Criterion Stain Free™ SystemThe Criterion Stain Free system comprises the ChemiDoc™

ChemiDoc MP System with Image Lab Software 106• Elimination of the need for acetic acid and methanol in staining and destaining, which reduces organic

Imaging Gels107Imaging GelsUse unstained standards with stain-free gels, as some prestained standards are not detected by the stain-free system. To mo

ChemiDoc MP System with Image Lab Software 108

ChemiDoc MP with Image Lab Software109Appendix B. TroubleshootingFollow these suggestions to troubleshoot your ChemiDoc™ MP system.Problem Possible Ca

ChemiDoc MP with Image Lab Software | Introduction2• Smart, application-based protocol setup using Image Lab software, which assists you by presenting

ChemiDoc MP with Image Lab Software | Appendix B. Troubleshooting110Printout does not look like the monitor image• Monitor settings are wrong • Printe

ChemiDoc MP with Image Lab Software111Appendix C. AccessoriesCalibrating AccessoriesIf you are installing accessories along with your original system

ChemiDoc MP with Image Lab Software | Appendix C. Accessories112UV/White Light Conversion ScreenThis optional white light conversion screen (catalog #

Installing Optional Accessories113The XcitaBlue conversion screen is held in place by adhesive-backed edge guides. After the edge guides are installed

ChemiDoc MP with Image Lab Software | Appendix C. Accessories1146. Center the gel on top of the XcitaBlue conversion screen, and proceed with normal i

Installing Optional Accessories1152. Place the gel alignment template that matches the size of your sample tray or agarose gel into the magnetic locat

ChemiDoc MP with Image Lab Software | Appendix C. Accessories116CALIBRATING THE SYSTEM TO USE THE ORANGE FLUORESCENCE REFERENCE PLATE Your system must

Ordering Information117Ordering InformationThe following table contains catalog numbers and descriptions for all parts available for the ChemiDoc MP s

ChemiDoc MP with Image Lab Software | Appendix C. Accessories1181001-4106 Thumbscrew for camera170-8081 Filter, standard emission, 62 mm100-1370 UV bu

Ordering Information119170-8354 EZ Load 500 base pairs molecular ruler170-8355 EZ Load 1 kb molecular ruler170-8205 2.5 kb molecular ruler170-8200Ampl

System Components3Image Lab SoftwareThe system ships with a full version of Image Lab software. In addition to controlling the imager system, image ca

ChemiDoc MP with Image Lab Software | Appendix C. Accessories120

ChemiDoc MP with Image Lab Software121Appendix D. Mitsubishi P93/P95 Thermal Printer SettingsWindowsThe printer driver can be found on the Image Lab™

ChemiDoc MP with Image Lab Software1224. Configure the correct paper size: select 1280 x 1280.5. Click OK to apply your changes.MacThe printer driver

Mac1233. In the Settings drop-down list, select Page Attributes.4. In the Format For drop-down list, select the Mitsubishi printer.5. In the Paper

ChemiDoc MP with Image Lab Software124

Appendix E: Regression Calculation Methods125Appendix E. Regression Calculation MethodsEach regression method calculates a standard curve. Some of the

ChemiDoc MP with Image Lab Software | Appendix E. Regression Calculation 126Logistic: The logistic-4PL equation iswhere:x = mobilityy = molecular weig

ChemiDoc MP with Image Lab Software127GlossaryCCD: (Charge-coupled device) A light-sensitive silicon chip used as a photodetector in ChemiDoc™ MP and

ChemiDoc MP with Image Lab Software | Glossary128Quantitative imaging: Determines the quantity of a protein’s components through analysis of the pixel

ChemiDoc MP with Image Lab Software129IndexAAbsolute quantity 79Absolute volume 89Accessories 111Activation optionsStain Free gel 107Adding annotation

ChemiDoc MP with Image Lab Software | Introduction4Optional AccessoriesBio-Rad Laboratories, Inc. offers a selection of optional filters and illuminat

ChemiDoc MP with Image Lab Software | Index130Crop tool 70Cubic splinedefined 126Cubic spline regression method 80Custom application 25, 43creating 25

ChemiDoc MP with Image Lab Software131Image Lab software 3Image tools 70Image Transformset image contrast 52Instrument calibration wizard 9LLanebackg

ChemiDoc MP with Image Lab Software | Index132Relative volume 88Relative volume quantity 89Replacingfuse 103UV starter 101UV transilluminator 101Repor

ChemiDoc MP with Image Lab Software133ZZoom toolsresizing images 51

Life ScienceGroup 0711 Sig 021110022469 Rev BBio-Rad Laboratories, Inc.Web site www.bio-rad.com USA 800 424 6723 Australia 61 2 9914 2800 Aus

System Requirements5System RequirementsImage Lab software runs under Windows XP Professional, Windows 7, and Mac OS X. Images scanned at high resoluti

ChemiDoc MP with Image Lab Software | Introduction6NUCLEIC ACID GELSPROTEIN GELSBLOTS• Ethidium bromide • GelGreen• SYBR® Green•GelRed• SYBR® Safe• Fl

ChemiDoc MP Technical Specifications7ChemiDoc MP Technical Specifications ApplicationsChemiluminescence YesFluorescence* YesColorimetry/densitometry Y

ChemiDoc MP with Image Lab Software | Introduction8ChemiDoc MP Imager WorkflowFollowing are the basic steps to acquiring, analyzing, and archiving an

ChemiDoc MP with Image Lab Software92 System CalibrationWhen your system is installed, system calibration is performed with a calibration wizard. See

Copyright ©2011 Bio-Rad Laboratories, Inc. Reproduction in any form, either print or electronic, is prohibited without written permission of Bio-Rad L

ChemiDoc MP with Image Lab Software | System Calibration10The Instrument Setup dialog box available from the Edit menu displays these calibration resu

ChemiDoc MP with Image Lab Software 113 Image Lab™ Software OverviewImage Lab image acquisition and analysis software works with the ChemiDoc™ MP imag

ChemiDoc MP with Image Lab Software | Image Lab™ Software Overview12 Interface OverviewThe following illustration shows the Image Lab software main w

Interface Overview13ToolbarMany Image Lab software tools can be selected by clicking toolbar icons. The Screenshot tool enables you to send a screen c

ChemiDoc MP with Image Lab Software | Image Lab™ Software Overview14Display ToolboxThe display toolbox near the top of every image enables you to dis

Interface Overview15Analysis Tool BoxThe Auto Analysis button quickly analyzes images. The remaining tools customize the analyzed data.Note: An imag

ChemiDoc MP with Image Lab Software | Image Lab™ Software Overview16Quantity Tools enable you to automatically quantitate bands, using either relativ

Interface Overview17Recent Images gives you the choice of opening a recent image file.Recent Protocols gives you the choice of opening a recent protoc

ChemiDoc MP with Image Lab Software | Image Lab™ Software Overview18• Volume Table to File—exports as a CSV file so your volume table can be opened i

Interface Overview19• The Volume Table tab enables the researcher to choose appropriate identifiers for the Volume Table and provides the option of ex

ChemiDoc MP with Image Lab SoftwareiiiSafety and Regulatory ComplianceImportant Safety InformationPlease read these instructions before attempting to

ChemiDoc MP with Image Lab Software | Image Lab™ Software Overview20HELPImage Lab Help displays the help system. User Guide displays the instruction

ChemiDoc MP with Image Lab Software214 Acquiring ImagesImage Lab software can run specific applications with repeatable workflows using custom protoc

ChemiDoc MP with Image Lab Software | Acquiring Images22The Protocol Setup WindowThis is the Protocol Setup window for a single-channel protocol. The

The Protocol Setup Window23The Multichannel Protocol Setup window has some different settings, but you work with it the same way as the Single-Channel

ChemiDoc MP with Image Lab Software | Acquiring Images24Creating a Single-Channel ProtocolSetting up a single-channel protocol consists of three proce

Creating a Single-Channel Protocol25Note: For a list of applications with all required detection reagents, light sources, and any conversion screens o

ChemiDoc MP with Image Lab Software | Acquiring Images262. Choose a unique application name. 3. Select a light source, filter, and image color on the

Creating a Single-Channel Protocol27prominent bands. Exposure time can range from 0.001 to 7200 seconds with the ChemiDoc™ MP system After imaging a g

ChemiDoc MP with Image Lab Software | Acquiring Images28In this example, the bar in the Signal Accumulation Setup dialog box shows that images will be

Creating a Single-Channel Protocol29Analyze ImageSTEP 2. DETECT LANES AND BANDSTo analyze the gel or blot, Image Lab software must detect lanes and ba

ChemiDoc MP with Image Lab Software | Safety and Regulatory ComplianceivGeneral Precautions• Please read the instruction manual carefully• The instrum

ChemiDoc MP with Image Lab Software | Acquiring Images30STEP 3. ANALYZE MOLECULAR WEIGHT10. To analyze molecular weight automatically, select Analyze

Creating a Single-Channel Protocol31STANDARD LANES12. Choose which lane contains your standards by typing lane numbers or the words First and Last in

ChemiDoc MP with Image Lab Software | Acquiring Images32REGRESSION METHODSA regression method is used to calculate the molecular weight of the unknown

Creating a Single-Channel Protocol33Output SettingsImage Lab software prints to the default printer unless you select otherwise.You have three choices

ChemiDoc MP with Image Lab Software | Acquiring Images34Review Protocol Settings16. Click Protocol Summary in the left pane of any Protocol Setup wind

Creating a Multichannel Protocol35Creating a Multichannel ProtocolSetting up a multichannel protocol consists of two parts:• Choosing acquisition sett

ChemiDoc MP with Image Lab Software | Acquiring Images36APPLICATION OPTIONS1. In the right pane of the Protocol Setup window, click Configure in the C

Creating a Multichannel Protocol373. Choose Image Exposure Time—if you select the top radio button, Image Lab software acquires an image estimating an

ChemiDoc MP with Image Lab Software | Acquiring Images38Rather than manually acquiring a series of independent images with different imaging times, SA

Creating a Multichannel Protocol39Channel Color—select a color to display the sample image. Assigning each channel a different color makes it easy to

vFUSESThe universal hood of the ChemiDoc MP imaging system has two user-serviceable fuses, F1 and F2, which are located on the bottom rear panel and a

ChemiDoc MP with Image Lab Software | Acquiring Images40STEP 4. SPECIFY OUTPUT7. Select Specify Output in the left pane of the Protocol Setup window t

Choosing a Protocol41You can also choose recently used protocols or image files from the lists on the Start Page.Editing ProtocolsProtocols can be cha

ChemiDoc MP with Image Lab Software | Acquiring Images42Position GelTo position a gel:1. Click the yellow Position Gel button in the Protocol Setup wi

Setting Up a Custom Application43The software runs through the steps in imaging mode, imaging each channel. A message at the bottom of the screen indi

ChemiDoc MP with Image Lab Software | Acquiring Images44To create a custom application:1. Select New in the Manage Custom Applications dialog box.The

Application Tables45Application TablesThe following tables list the applications and primary filters for each light source.Base System—UV, Standard Fi

ChemiDoc MP with Image Lab Software | Acquiring Images46Red Epi (catalog #170-8283) 695/55 FilterCY5 DyLight 650CY5.5 DyLight 680Alexa 647 IRDye 680Al

Creating a Multichannel Image from Single Images47Creating a Multichannel Image from Single ImagesYou can create a multichannel image from existing si

ChemiDoc MP with Image Lab Software | Acquiring Images48

ChemiDoc MP with Image Lab Software495 Viewing ImagesOnce a gel has been imaged, the image appears in the workspace. Many controls are available to op

ChemiDoc MP with Image Lab Software | Safety and Regulatory Compliancevi

ChemiDoc MP with Image Lab Software | Viewing Images50Displaying Gel ImagesNotice the display toolbar buttons above the gel image. Each of these tools

Displaying Gel Images51•Volume• Absolute Quantity• Relative Quantity•Band %•Lane %VOLUMESIf you have drawn volume boundaries on the gel, you can displ

ChemiDoc MP with Image Lab Software | Viewing Images52Image Transform Use the Image Transform dialog box to adjust image brightness and contrast, opt

Displaying Gel Images53• Highlight saturated pixels—when this checkbox is selected, areas of the image with saturated signal intensity (above a measur

ChemiDoc MP with Image Lab Software | Viewing Images54Image Colors You can choose a colormap for your image results file. Viewing the image with a dif

Displaying Gel Images553-D Projection Clicking the 3-D button transforms the gel image into a solid 3-dimensional model spinning in space with x, y, a

ChemiDoc MP with Image Lab Software | Viewing Images56IMAGE DETAILSAcquisition and image information appear in this tab.Note: For multichannel images,

Displaying Multichannel Images57In the display toolbox above the image, additional controls are available for viewing the multichannel image.Multichan

ChemiDoc MP with Image Lab Software | Viewing Images58You can also specify the channels to include in the merged view using the drop-down list.Splitti

Displaying Multichannel Images59CHANGE LAYOUT You can choose a layout for the image panes. Clicking Change Layout shows a list of display options for

viiContentsChapter 1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1ChemiDoc™ MP Imaging System. . . . . . . . . . .

ChemiDoc MP with Image Lab Software | Viewing Images60Horizontal ViewSingle ViewDisplaying Data The results from analyzed data associated with the gel

Displaying Data61Analysis Table Options Numerical data associated with an analysis can be viewed in an analysis table. Data from the Lane and Band ana

ChemiDoc MP with Image Lab Software | Viewing Images62Export formatting—select checkboxes to include lane headers (Lane and Band table tab only) and/o

Displaying Data63Abs. Quant.—absolute quantification of the band.Rel. Quant.—relative quantification of the band compared to the reference band.Band %

ChemiDoc MP with Image Lab Software | Viewing Images64Lane Profile The Lane Profile option shows a cross-section of the selected lane, rotated 90°. Yo

Displaying Data65IDENTIFY BANDS BYYou can display callouts on the Lane Profile to indicate these attributes:• Band Number• Band Label• Molecular Weigh

ChemiDoc MP with Image Lab Software | Viewing Images66Standards appear in green; unknown bands appear in red. You can toggle the molecular weight disp

ChemiDoc MP with Image Lab Software 676 Analyzing ImagesAnalysis Toolbox tools are enabled once an image file is opened and in focus. An active or “in

ChemiDoc MP with Image Lab Software | Analyzing Images68DETECTION SETTINGSBand detection sensitivity options are as follows:Low Band Detection Sensit

Analysis Toolbox Tools69MOLECULAR WEIGHT ANALYSIS SETTINGSMolecular Weight Standard—choose any of the many Bio-Rad standards or other standards you ha

ChemiDoc MP with Image Lab Software| ContentsviiiOutput Settings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

ChemiDoc MP with Image Lab Software | Analyzing Images70Image Tools To show the image tools menu, click Image Tools. FLIP You can flip the gel image

Analysis Toolbox Tools714. (Optional) Type a name for the crop settings in the dialog box that appears and click OK.5. Right-click and select Crop or

ChemiDoc MP with Image Lab Software | Analyzing Images72Lane and Band ToolsClick Lane and Band Tools to select a tab, one for the lane tools and one

Analysis Toolbox Tools73ALL LANESResize—you can resize all lanes by dragging the handles on the corners of the red square to fit the gel image.Adjust—

ChemiDoc MP with Image Lab Software | Analyzing Images74A large disk follows the profile trace less closely, touching fewer points along the trace an

Analysis Toolbox Tools75Detect Bands—this option opens a dialog box in which you can select band detection sensitivity and the lanes to which it appli

ChemiDoc MP with Image Lab Software | Analyzing Images76Molecular Weight Analysis ToolsMolecular Weight Analysis Tools enable you to determine molecu

Analysis Toolbox Tools77Click Change to access the Manage Standards dialog box, where you can choose another standard or add third-party standards. ST

ChemiDoc MP with Image Lab Software | Analyzing Images78To select a reference band from an image and quantitate all other bands relative to it:1. Cli

Analysis Toolbox Tools79ABSOLUTE QUANTITY TABAbsolute quantification is used to quantify bands based on known standard bands using a calibration curve

ixVolume Tools . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 85Chapter 7 Generating Reports . . . . . . .

ChemiDoc MP with Image Lab Software | Analyzing Images803. Select a unit of measure from the choices listed in the Units drop-down list. 4. Select a

Analysis Toolbox Tools81represented by green triangles. Unknown values are represented by red triangles.Note: Selecting the Force Through Origin check

ChemiDoc MP with Image Lab Software | Analyzing Images82Annotation ToolsYou can annotate results with text and arrows to draw attention to areas of i

Analysis Toolbox Tools83Arrow—to add an arrow, first click Arrow. Click the area where you want the arrow to start and then drag to stretch the arrow

ChemiDoc MP with Image Lab Software | Analyzing Images84Foreground—click a text annotation or arrow. This activates the Foreground field, so you can

Analysis Toolbox Tools85Volume ToolsVolume tools enable you to manually quantitate features on a sample image when automated lane and band analysis is

ChemiDoc MP with Image Lab Software | Analyzing Images86A default label appears within the shape drawn. The volume label can be one of three types—U

Analysis Toolbox Tools87VOLUME TYPESYou can define the volume type (unknown, standard, or background), the quantity of standard volumes, or enter a cu

ChemiDoc MP with Image Lab Software | Analyzing Images88Local—local background subtraction calculates a separate background intensity for each unknow

Analysis Toolbox Tools89Relative quantities are displayed in the Relative Quantity column in the analysis table (Volume Table tab). The relative quant