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USER GUIDE USER GUIDE

1. cccccccscceecesescceseceeseeenees 35 SPECIFICATIONS uu cccecceccececceccececceccuseccueeneeseueeeneeaneees 36 WARNINGS PRECAUTIONS amp NOTICES Throughout this manual the following types of notifications require your close attention A WARNING This type of notification tells how to avoid serious personal injury IMPORTANT This type of notification tells how to avoid damaging the microscope and or voiding your warranty contact your local distributor for more warranty information This symbol indicates information specific to the color camera version MONOCHROME CAMERA VS COLOR CAMERA The EVOS fl microscope is factory configured with either a monochrome camera or a color camera Monochrome cameras are commonly used for high performance fluorescence applications and provide the best sensitivity for detection of faint fluorescence signals Color cameras have lower fluorescence sensitivity but have the advantage of being able to differentiate structures by color in transmitted light e g imaging stained tissue samples Throughout this User Guide any operational differences between the two versions of the microscope have been noted 2012 Advanced Microscopy Group All rights reserved Doc Control Number ZP PKGA 0495 REV C6 Published 4 JUN 2012 STANDARD ITEMS INCLUDED Before setting up your new EVOS unpack the unit and accessories and verify all parts are present Contact your distributor if anythin
2. EVOS USER GUIDE Digital Inverted Microscope for Fluorescence and Transmitted Light Applications mercury free w energy efficient Sas EVOS USER GUIDE SETUP r E A EE 3 Standard Items INCIUG d cc ecsesseeeeseeseeeeneeeeeeeeseaaes 3 Moving Transporting EVOS ccccccccsssssssseeceeeeeeseeseees 4 Operating ENVirOnMent cccccsssseesesesseeseceesseeneeeeeeaues 4 Mechanical Stage u ccsscssscsssssessseseeeeseseeasseeeeueseeenaeeeseas 4 LED Light Cubes uuu ccceseeesesseseeeseeeeseeeeaeseseeusseenaeeeeeas 5 Power SUPDLY uu ccscsesecsesseeeseseseeuseeeenseeenaeueeeeueeeeenaeeeseas 5 USB POTUS scarcer csanestsciedusueupeatdedenteusustesauiedenseneneversudedesteunin 5 DVI Output PONE sieccestwedticecusececdeceicicutvenemateuediiercueitedduvtes 5 SNE ete aterctecitutettieccenfeeewo senda EEEE 6 Installing EVOS in a Cell Culture Hood ccccceee 7 QUICK REFERENCE DIAGRAMS ce ececceeeeeeeseeeeeeeees 8 BASIC OPERATION Vu cceccceceesecesseeeeceseeeeecneeeeeeesseeeeenes 9 Fluorescence Operation cccssesssseesssessseseeeeseeeneeeenes 9 Brightfield or Phase Contrast Operation 0008 11 ADVANCED OPERATION uu ccccescceecceeseeeeeneeeeeeeneeenees 13 Logging In Creating New User LOoginS sssss0008 13 Saving Images amp Working with Files 14 Using the QuickSave Option cccccseessseeseseeeeeeeeeeens 15 Recording Time Lapse IMAGES
3. P DOMAIN_E Windows SMB Network with available domains and servers EVOS IP Address 192 166 4 71 DHCP Server 192 168 4 1 Windows SME Network Y DOMAIN_A Server_ Y Server_2 Shared_Folder_1 O shared Fle Shared_Folder_3 Domain User N rg as assword DOMAIN A UserName Server_2 Shared_Folder_2 E c Server_2 Shared_Folder_1 lt Q Server_2 Shared_Folder_2 Remove o Adding shared folders The Network page only displays the top level folders View subfolders in the Browse popup when saving a file Basit Quick Save Channels Network Servite OuickSave File BROWSE Folder New Folder W NETWORK Server_1 Shared_Folder_1 P Sub_Folder_ P Sub_Folder_2 gt Sub_Folder_3 gt NETWORK2 Server_1 Shared_Folder_2 gt USBI Cancel Network destinations in the QuickSave Browse popup www amgmicro com Connect EVOS to a Network continued The upper list box of the Network page will display the top level available domains of the Windows SMB network file tree Click the triangle icon or double click the domain name to expand a domain and display the available servers Note If a domain server or shared file appears on the file tree without a triangle icon and you are not able to expand or open it your permission to access that item Is restricted Enter your network domain user name and password in the login fields and select a server to view the top level of shared fold
4. Remaining Minutes 4 Disk Space 1950 MB Total Review 4 4 h a 15 a Pause Abort E 19 Find amp Focus Po RemainingMB 1765 OO review 4 4 a Play Abort E 19 Find amp Focus Time Lapse Progress paused www amgmicro com RECORDING TIME LAPSE IMAGES With EVOS you can set up your cells and program the microscope to record time lapse images To use this feature open the Time Lapse tool in the toolbar specify the settings and click Start You may pause or cancel sessions in progress START A TIME LAPSE SESSION 1 Once the specimen is focused and ready tighten the stage brakes to prevent the stage from drifting during the session 2 Open the Toolbar and expand the Time Lapse tool 3 Click the Interval text field and enter a value 4 Choose a unit of measure for the capture interval from the interval Unit drop down menu 5 Click the Duration text field and enter a value 6 Choose a unit of measure for the duration from the duration Unit drop down menu Note The Images field shows the total number of images for the session 7 To save the session in a folder other than the default location click the Browse button to select the destination 8 Inthe Browse popup shown on p 75 highlight the desired folder and click OK 9 Under File click the Base Filename text field to enter a name and then choose a file type png or tif from
5. cessseesesseeeeeeeeeeees 16 Using the Transfection TOol ccccccsscseeesssseeeeeeeeseneees 17 Counting Cells roesini 18 Reviewing IMAGES ccccceeceeeesesseseeneeseeeeeeseauseeeeneeeeens 19 Using Digital ZOOM ccccscesssssseeeeeeseeeesseeeesneseeenaeeees 20 Connecting EVOS to a Network ccccssssssssseesseenaes 21 Changing LED Light Cubes esssesseeesseeeeeeeeees 23 Updating Software ccccccccsssseceeeeecsseeeeeeenesseeeensaeeeees 24 Setting Date and Time ou cscccsecsesseeesessseeeeeeeeneeeeeees 25 CONTROLS GLOSSARY cccccecccesceusceessueccseccusecnsseusees 26 Onscreen Controls ccccececeecceecceecceecceceeceeeeeeseeeseeeeens 26 Mechanical Controls assasnasnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnn 30 CARE amp MAINTENANCE cccccecceccececcecssceccneeseeeenees 32 General CAV 6 sei sesesicxscwascasenarveuvcussedeseniecanecnnenianecieeweawens 32 Objective Lens Care ccsscsessssseseeseseeeeesseeeeseeeeneeeeees 32 Stag Se ene eee eee een ne nee een nee 32 Sterilization PrOCCCUIPCS cecceesceeeceeccneccnsceeeeeeeeeneeess 32 TROUBLESHOOTING w ccccceccecescecceccececcecenceccueeeseeeueees 33 Image Quality ISSUES cccseeesesseeseeceeseeeesseeeeneseesnaees 33 Software Interface ISSUCS ccseccceeeceeeeeceeeeeeeeneeeees 33 Mechanical ISSUES c ccceeceecceeceeeceeeceeeeeeseeescneeeeeeeeees 34 CUSTOMER amp TECHNICAL SERVICE 55 AMG Contact INformation
6. the File Type drop down menu 10 To create a video avi file select Write Video File 11 Click the Start button to begin the time lapse session EVOS will display the Time Lapse Progress popup as long as the session is active Note The Review slider lets you review the images already captured during the current session The Play button shows all the images in sequence PAUSE AND RESTART A TIME LAPSE SESSION In the progress popup click the Pause button to suspend the time lapse captures The progress popup will dim and a Resume button will replace the Pause button Alternatively to pause and adjust the settings click the Find amp Focus button Click the Start button to resume the time lapse capture sequence or uncheck the Resume radio button and start a new time lapse session ABORT A TIME LAPSE SESSION In the progress popup click the Abort button A dialog box will pop up giving you the option to delete or keep the files already saved Clicking Cancel will resume the session EVOS User Guide REV C6 Published 4 JUN 2012 lt C gt ADVANCED OPERATION WN GDN NEO Sequence paused between images www amgmicro com USING THE TRANSFECTION TOOL EVOS Transfection tool expedites the capture and overlay of images for transfection analysis 1 Choose a light cube focus on the sample and adjust the lighting See steps 1 13 of Fluorescence Operation p
7. 18 IMAGE REVIEW TOOL The Image Review tool allows you to review still images or play video files from the USB drive or network connection You may also use this tool to rename or delete saved files See Reviewing Images p 19 for more details Saree SETTINGS BUTTON 62 0 Count tool i The Settings button bottom center opens the a Settings dialog box Within the Settings dialog box Settings the following options are available button gt Basic Tab e This EVOS section displays the serial number and software version e Login section controls default Guest login e Display section controls the following options Highlight saturated pixels in red indicates overexposed areas onscreen Reset Scalebar Location moves the Scalebar back to its default position in the lower right corner of the screen Enable Mouse Wheel allows the mouse scroll wheel to control illumination levels Mouse Speed controls the mouse s reaction time LCD Backlight controls the LCD lighting e File Save section allows you to specify 8 bit as the default format for TIFF files Settings QuickSave Tab gt QuickSave Tab allows you to set up a file name Image Review Tool monochrome version shown color version shown count file type and folder to save captured images www amgmicro com automatically as well as the option to save each channel in a separate file for each c
8. 9 for detailed instructions Open the Toolbar and expand the Transfection tool Note The Pause after first image option allows you to adjust focus if necessary before capturing the transmitted light channel Click the Run Sequence button The sequence always starts with the fluorescence channel and finishes with the transmitted light channel Note The Lighting Override feature allows you to select a channel to activate with the LIGHT ON OFF button If the Pause option is selected adjust focus and click the Continue button If Pause is deselected go to step 5 The dual channel image will display with the Overlay tab selected Adjust the brightness and contrast settings as desired and save the image See Saving Images amp Working with Files p 14 for detailed instructions EVOS User Guide REV C6 Published 4 JUN 2012 zAV C gt ADVANCED OPERATION WN DN NEINO S COUNTING CELLS The Count tool streamlines cell counting by marking items with up to 6 labels onscreen As you tag items EVOS will keep a running tally of counts with percentages for each label assigned Document your results simply by saving the tagged image with the Count tool displaying the totals 5 2 Count ae Tags A Mode Add Delete Labels 1 Non Expres 44 62 9 2 Transfected 26 37 1 0 0 0 0 0 0 0 0 6 TOTAL 70 Clear All a 10 7 Setumgs Hide T
9. box where you can fine tune your live image Brightness Contrast Saturation and Hue prior to capture VIEW SCALEBAR OPTION The Scalebar option bottom center is a toggle button that displays or hides the Scalebar tool This option is only available after an image is captured To move the Scalebar simply click and drag it VIEW TOOLBAR OPTION The Toolbar option bottom center includes the Time Lapse Transfection Count and Image Review tools Click the small gray triangle to open each tool EVOS User Guide REV C6 Published 4 JUN 2012 CONTROLS GLOSSARY TOOLBAR Time Lapse tool SETTINGS Serial Number F1209 1441 026 oftware Revision 15844 Detaatt to Guest on startup Display Highlight sanmaced pixels inred R yul Caled Enabic Mouse Whee CU Gackhgitt Less Always save TIFF as 3 bir Save TIFF in Microsoft compatible format Settings Basic Tab TIME LAPSE TOOL The Time Lapse tool allows you to set up the interval duration and the file name destination for time lapse sessions See Recording Time Lapse Images p 16 TRANSFECTION TOOL 57 1 The Transfection tool automatically captures and ji overlays images for transfection analysis See Using the Transfection Tool p 17 sec COUNT TOOL The Count tool allows you to tag cells and other features using up to 6 labels and it calculates the totals and percentages for each label See Counting Cells p
10. delete all tags for all labels To save an image showing the labels counts and percentages as shown in the Count tool select the Save Image with Toolbar option and click the Save button Deselecting this option will produce an image saved with the tags only EVOS User Guide REV C6 Published 4 JUN 2012 lt 5 ADVANCED OPERATION ADVANCED MICROS CAPTURE PLAY rw n mpn r r Ole G a s J tocar Image Review Tool View rename or delete a file Guest Pind amp Pocus bt Se o herme VIRTUAL KEYBOARD Rename File Cells awT Cells avi Backspace Clear za Date o Caps Lock Accept Cancel Virtual keyboard CONFIRM FILE DELETE Please confirm that you wish to permanently delete the Cells avi file Yes No Confirmation popup www amgmicro com REVIEWING IMAGES The Image Review tool allows you to review still images or play video files from the USB drive or network connection You may also use this tool to rename or delete saved files 1 Open the Toolbar and expand the Image Review tool 2 The preview list displays thumbnail images for all viewable files in the selected directory the top level USB directory is selected by default If there are no viewable files in the directory the preview area will be empty Note The File Type drop down menu filters files by type By default it is set to display all files with png tif jog bmp or avi exte
11. digital image 4 Right click to restore the view to unzoomed magnification Note Capturing and saving a zoomed image will result ina file showing the actual magnification not the zoomed magnification If the scalebar is active it will appear in the file EVOS User Guide REV C6 Published 4 JUN 2012 lt 5 ADVANCED OPERATION ADVANCED MICROSC CONNECTING EVOS TO A NETWORK You can log EVOS onto a Windows SMB network via an Ethernet cable connection and save captured images directly to shared folders on the network Note SMB is the only supported protocol No other protocols such as HTTPS FTP or WebDAV are currently supported If you are connecting to a Linux server it will have to use Samba in order for EVOS to find it Contact your network administrator for help if a physically connected EVOS cannot find the SMB network ITEMS NEEDED FOR ETHERNET CONNECTION To set up the network connection you need the following items not included gt Ethernet cable gt USB to Ethernet adapter LOGGING ONTO THE NETWORK Guest Find Focus LIGHT u 1 Verify the microscope is powered on and the network Actual Yi F cable is plugged into the correct jack and connected AMG E via the adapter to the USB port poe PULTON ASEE TO GUESERTONIE 2 Be sure you are logged in under your own EVOS user ID The current user ID is displayed above the AMG logo in the bottom left corner of the screen See Logging In V
12. each objective periodically or when necessary with an optical grade swab and a pre moistened lens wipe or lens paper moistened with lens cleaning solution To avoid scratching the soft lens coatings use only optical grade cleaning materials and do not rub the lens The Objective Lens Cleaning Kit AMEP 4702 is available for use with all your objective lenses Note To protect all optical components of the microscope use the dust cover when the microscope is not in use STAGE CARE gt Clean the stage as needed with paper towels or Kimwipes dampened with 70 ethanol gt When moving EVOS be sure to lock the stage with the stage lock pin as shown on p 4 to prevent the stage from sliding STERILIZATION PROCEDURES To sterilize the EVOS please follow these procedures 1 Turn power OFF 2 Clean the LCD display a Use a soft dry lint free cloth to wipe off any dust from the screen b Clean the LCD display with a non alcohol based cleaner made for flat panel displays IMPORTANT Do not spray cleaning fluid directly onto the screen as it may drip into the display or optics 3 Lightly wipe EVOS working surfaces stage top focusing knobs objective selection wheel housing with paper towels or Kimwipes dampened with 70 ethanol or 4 000 ppm hydrogen peroxide H 0 IMPORTANT Do not allow sterilization solution to get into lubricated areas such as the stage roller bearings or any points of rotation such as axles for th
13. next image acquired in the customized channel EVOS User Guide REV C6 Published 4 JUN 2012 lt 5 ADVANCED OPERATION ADVANCED MICROSC VIEW SETTINGS SAVE Toolbar tt a Settings button O Intensity INFO Max 144 Ratio 14 40x Min 0 Cursor 0 Objective SETTINGS Basic Quick Save Channels Network Service CONTACT YOUR DISTRIBUTOR FOR SERVICE Microscope Turret Position Objective Lens AMG 2X Plan LWD AMEP 4601 AMG 4X Plan LWD PH AMEP 4602 AMG 10X Plan FL AMEP 4623 AMG 20X Plan FL AMEP 4624 AMG 40X Plan LWD PH AMEP 4635 Numbered Turret Positions Hot Pixel Correction Software Update Correct Update Error Log Date and Time Send Change Cancel Settings Service Tab The EVOS update file was not found on a USB flash drive The update cannot proceed sorry Missing update notification Verifying and Unpacking Update Verification progress bar LAP DATE Update confirmation popup EVOS Software Updater DO NOT TURN OFF POWER TO THE UNIT OR PLUG OR UNPLUG DEVICES TO THE USB PORTS WHILE THE UPDATE I5 IN PROGRESS Sibu U pirang app bain bootloader Installation progress bar www amgmicro com UPDATING SOFTWARE Periodically AMG adds functionality and other improvements to the EVOS user interface We recommend keeping your EVOS microscope up to date with the latest software If you have any questions about software updates cont
14. to start the update 5 The screen will display update progress When the update is complete the main EVOS screen will reappear IMPORTANT Do not power off unplug the USB stick or add remove any devices during the update EVOS User Guide REV C6 Published 4 JUN 2012 lt 5 ADVANCED OPERATION ADVANCED MICR Q VIEW SETTINGS SAVE intensity INFO T OES Max 144 Ratio 14 40x Toolbar iat Min 0 Cursor 0 Objective Settings button O SETTINGS Basic Quick Save Channels Network Service CONTACT YOUR DISTRIBUTOR FOR SERVICE Microscope Turret Position Objective Lens AMG 2X Plan LWD AMEP 4601 AMG 4X Plan LWD PH AMEP 4602 AMG 10X Plan FL AMEP 4623 AMG 20X Plan PL AMEP 4624 AMG 40X Plan LWD PH AMEP 4635 Numbered Turret Positions Hot Pinel Comection Software Update Comect Update Eror Log Date amd Time Send Change B 2 O OK Cancel Settings Service Tab Date and Time Time 02 10 PM Date Tuesday January 04 7011 Hours Minutes 7 PM F Date Tuesday January 04 2011 Time zone GMT 08 00 Pacific Time US Canada 3 OK Cancel Date and Time popup www amgmicro com SETTING DATE AND TIME To ensure accurate date and time information on saved image files verify the EVOS date time settings before your capture session 1 Click the Settings button to open the Settings dialog box and then select the Service tab Note Cha
15. IEW SETTINGS SAVE intensity INFO Creating New User Logins p 13 for more information Blot ai heal ia 3 Click the Settings button to open the Settings dialog box and then select the Network tab Toolbar Settings button Note EVOS will try to connect for about 30 seconds ae If there is a problem with the connection the Network page will display No Items Basic Quick Save Channels Network Service Double check the physical connections and Pa lt O click the Refresh Network button During the F Use Static IP Address refresh a progress icon will appear Unless there is an issue with the network or you are using an incompatible adapter refreshing the connection should resolve the problem No Items within a few moments Contact your network administrator for help if the problem persists IP address mot assigned ea a Name A If your configuration requires using a static IP address select the Use Static IP Address option enter your IP Address Subnet Mask Gateway nae Address and DNS Server Address then click Set Static IP No tems Remove OK Cancel Refreshing the network connection continued on next page www amgmicro com EVOS User Guide REV C6 Published 4 JUN 2012 lt 5 ADVANCED OPERATION ADVANCED MICROSC EVOS IP Address 192 163 4 71 DHCP Server 192 168 4 1 Windows SMEB Network DOMAIN_A Server_ Server_2 Server_s 0 server_4
16. These locations will also be available in the Save dialog box and through the Browse button in the Time Lapse and Image Review tools EVOS User Guide REV C6 Published 4 JUN 2012 zAV C gt ADVANCED OPERATION DVANCED MICROS Remove LED light cube access Light cube tool under cover under stage access cover S3 i gt Hlp Light cube tool attached to light cube VIEW SETTINGS intensity INFO Max 144 Ratio 14 40x Toolbar Min 0 Cursor 0 __ Objective Settings button Basic Quick Save Channels Network Service Custom Channel Settings TRANSMITTED E omy am DEFAULT DAPI F Cancel Assigning custom channel display colors not applicable for the color camera version www amgmicro com CHANGING LED LIGHT CUBES INSTALL LIGHT CUBE Optional LED light cubes are available contact info amgmicro com for details Each LED light cube is coded so EVOS will automatically recognize it in any position A A WARNING UV LIGHT HAZARD This microscope uses a Class 3B ultraviolet LED for the DAPI channel Before changing LED light cubes power off the microscope 1 Turn the power switch to the O OFF position before removing or changing light cubes 2 Move the stage back to allow access to the light cube access cover centered under the back of the stage 3 Loosen the thumbscrew to remove the light cube access cover and remove the light cube t
17. act your local distributor If you do not have your distributor information you can look it up at the or contact AMG Customer Service see p 35 DOWNLOAD SOFTWARE UPDATE Software updates are available under the Support menu at the 1 Download the update directly to the top level of a USB flash drive with at least 30MB available Do not open or rename the file on your computer EVOS will verify and install it during the update process 2 Download the current user guide for EVOS from the AMG website The updated user guide covers the new software functionality when features are added Alternatively you can get the latest software and documentation updates from your local EVOS distributor INSTALL SOFTWARE UPDATE 1 Plug the USB stick into the data port on the lower right side of the EVOS support arm 2 Click the Settings button to open the Settings dialog box and then select the Service tab Note Changing settings in the Service tab will affect the microscope s performance If service beyond the software update is needed please contact your EVOS distributor 3 Click the Update button in the Service tab A verification progress bar should appear If a missing update notification pops up be sure the USB with the update file is fully plugged in Click OK and then click the Update button again 4 After file verification an update confirmation dialog will pop up Check the revision details and click Yes
18. ags Save image with Toolbar 8 Show Grid ux25u Ww Count tool Settings Hide Tags Save image with Toolbar Show Grid ux2u 3 25u x 25u 0u x 50u 10u x 100u j F image Rewiew Detail of grid size menu and Show Grid option Selected tag drag to move www amgmicro com Acquire an image See BASIC OPERATION p 9 for detailed instructions Open the Toolbar and expand the Count tool Click in a black Label text field to name a label You may use up to 6 labels Under Settings you may choose a grid size in the drop down menu or leave the Show Grid option inactive Select a Label button and left click at each point onscreen to tag the items for that category Switch labels as desired EVOS will tag for the selected label Note To use Digital Zoom while counting cells first suspend the Count tool Either select the Hide Tags setting 8 click the triangle to minimize the Count tool or click the Toolbar option to minimize the whole Toolbar When the Count tool is suspended the left mouse button will behave according to the rules described in Using Digital Zoom p 20 After zooming reactivate the Count tool by deselecting Hide Tags or reopening the tool To move a tag select and drag it Left click anywhere else onscreen to deselect To delete a tag right click it or choose the Delete mode and left click it You may also use the Clear All button to
19. amp Focus LIGHT Actual On LIGHT ON button in the control bar Pa Light shield box in place on stage www amgmicro com The EVOS microscope uses both mechanical and software controls for operation Mechanical controls include the stage X Y axis knobs focusing knobs objective selection wheel and the light cube selection lever Software controls are located in the control bar at the bottom of the screen The channel indicator bar at the top of the screen shows the selected filter cube or transmitted light position The login button displays the current user ID Refer to the QUICK REFERENCE DIAGRAMS p 8 as needed See also the CONTROLS GLOSSARY p 26 for more details FLUORESCENCE OPERATION 1 Turn on the microscope using the power switch on the right side of the microscope base 2 Plug a USB flash drive into one of the USB ports on the right side of the microscope arm 3 Place the sample on the stage using a vessel holder if needed 4 Set the magnification using the objective selection wheel on the front of the microscope 5 Move the light cube selection lever on the left side of the microscope all the way toward you the channel bar will highlight the Transmitted position 6 Turn on illumination using the LIGHT ON button located on the left side of the control bar 7 Focus the sample using the focusing knobs 8 Place the light shield box on the stage ov
20. an work with the same EVOS microscope without having to reset their preferences To use this feature set up a user profile for each regular user You may also assign user IDs for experiments in progress Note User profiles are not password protected All users should verify they are logged in correctly to avoid changing others settings LOG IN WITH AN EXISTING PROFILE 1 Click the login button at the bottom left of the screen this is the AMG logo with the current user profile indicated above 2 Select the desired user profile and click OK Note No password is necessary to log in ADD OR REMOVE A USER PROFILE 1 Click the login button O 2 To copy an existing profile highlight the profile in the user list select the Copy from name option and click the Add button The virtual keyboard will pop up so you can name the new profile 3 To create anew user profile without copying any settings deselect the Copy from option 9 click the Add button 8 and enter a user name 4 After adding a new user profile click OK to log in under that name and adjust settings as desired When you switch off EVOS will save your settings to memory 5 To remove a user profile highlight it and click the Remove button A confirmation dialog box will pop up Deleting the user profile will remove all its associated settings from memory 6 To rename a user profile click the Rename button and e
21. aptured image gt Channels Tab not shown allows you to assign a custom display color to each fluorescence channel monochrome camera version only See Assign Custom Channel Display Colors p 23 gt Network Tab not shown allows you to set up connections to shared folders via Ethernet See Connecting EVOS to a Network p 21 gt Service Tab not shown allows for maintenance functions contact your distributor if service is needed EVOS User Guide REV C6 Published 4 JUN 2012 CONTROLS GLOSSARY Save button SAWE FILE Save File dialog box VIRTUAL KEYBOARD New Folder Caps Lock Cancel Virtual keyboard Intensity INFO Max 915 Ratio 91 Min 0 Cursor 16 40x Objective i 5 QUICKSAVE Fluor RGB 0O0T 1if 40x Objective Info display variations c Note In all tabs the color camera version displays the QuickSave option instead of the Info bar 2X oom J 40x Objective Objective Objective turret Digital Zoom between positions Value display www amgmicro com SAVE BUTTON The Save button bottom right saves the current image to the USB flash drive or network folder A red USB icon indicates that the current image has not been saved yet After a file is saved the USB icon turns green SAVE FILE DIALOG BOX The Save File dialog box popup allows options for naming and filing captured images The following options are available in the Save Fil
22. back movements to position the specimen within the field of view particularly at high magnifications To secure the stage in place for time lapse image captures tighten the Y axis stage brake FOCUSING KNOBS Use the focusing knobs to bring the specimen into focus OBJECTIVE SELECTION WHEEL Turn the objective selection wheel to change magnifications The objective turret will click into place at each position EVOS User Guide REV C6 Published 4 JUN 2012 lt 5 CONTROLS GLOSSARY NWV NN GDN EINO s LIGHT CUBE SELECTION LEVER Move the light cube selection lever to change light channels The lever will click into place for each of the following positions gt Position O closest to front gt Position second from front gt Position second position from back gt Position furthest position to the back Note The channel indicator bar across the top of the screen displays the active channels for each lever position see p 26 PHASE ANNULI SELECTOR Set the phase annuli selector to the position that corresponds with your selected objective for phase or brightfield observations The selector will click into place for each position EXAMPLES gt BF use for brightfield observations gt 20 40 PH use for phase observations at 20x or 40x CONDENSER SLIDER SLOT Transmitted Light Applications Enhance image quality by inserting an optional slider all the way into the slot se
23. be used for low magnification 2x or 4x brightfield applications to improve flat field illumination 8 Turn on illumination using the LIGHT ON button located on the left side of the control bar 9 Focus the sample using the focusing knobs Phase annuli selector Insert slider into condenser Find amp Focus LIGHT _92 orr Owerlay LIGHT ON button in the control bar continued on next page www amgmicro com EVOS User Guide REV C6 Published 4 JUN 2012 CALG BASIC OPERATION ADVANCED MICROSC Light shield box in place on stage cover shown in place CAPTURE 6 LIGHT SAVE intensity INFO i Max 165 Save button 40x Objective Ratio 24 0 Cursor 12 AWE FILE VIRTUAL KEYBOARD Caps Lock Cancel Virtual keyboard www amgmicro com 10 1l 12 13 14 15 Brightfield or Phase Contrast Operation continued When switching from fluorescence to transmitted light with the light shield box on the stage remove the light shield box cover so the light from the condenser can pass through Adjust the illumination intensity if necessary using the Illumination slider on the control bar or the mouse scroll wheel Note Overexposed pixels will appear red Dim the illumination until the red highlights disappear to get the maximum level of brightness without any overexposed areas See p 28 for instructions on changing the o
24. butor If you do not have your distributor information you can look it up at the AMG website or contact AMG Customer Service see p 35 www amgmicro com EVOS User Guide REV C6 Published 4 JUN 2012 AMG CONTACT INFORMATION gt Toll free US amp Canada 866 614 4022 gt Local 425 368 0444 gt International 01 425 368 0444 gt Fax 425 368 0555 gt E mail customerservice amgmicro com gt Web site www amgmicro com Advanced Microscopy Group Customer Service business hours are 7 00 a m 4 00 p m Pacific Standard Time After hours you may leave a telephone message We will return your Call the following business day ADVANCED MICROSCOPY GROUP 22025 20th Ave SE Suite 100 Bothell WA 98021 Thanks for being our valued customer Advanced Microscopy Group is a Seattle area Bothell WA optical equipment design and manufacturing firm with a domestic and international client base Advanced Microscopy Group designs develops and manufactures optical systems and software for scientific biotechnology industrial and educational fields DISCLAIMER The information in this manual is furnished for informational use only and is subject to change without notice AMG assumes no responsibility or liability for any errors or inaccuracies that may appear in this document or any software that may be provided in association with this document No part of this document may be reproduced stored in a retrieval syste
25. cked into a position gt Capture image first scalebar is only available after capturing an image gt Click capture first It is only possible to save an image that is captured The USB icon on the Save button is red when there is a captured image to save gt Power EVOS off check the cable connections and power on the microscope gt Verify that the external display accepts DVI D input i e digital input A monitor with analog input only DVI A will not display the EVOS digital output gt Verify physical cable connections confirm the Ethernet jack is active gt Use a compatible network adapter gt If the physical connections are good and the problem persists contact your network administrator to resolve any network issues Note that a Linux network must use Samba for EVOS to be able to find it EVOS User Guide REV C6 Published 4 JUN 2012 lt S TROUBLESHOOTING MECHANICAL ISSUES LED light cube selection lever gt NEVER force lever does not move gt Remove LED light cube lock and replace with light cube access cover see p 5 Vessel does not sit securely on gt Use the correct vessel holder for the application refer to the EVOS Vessel Holders spec moving stage sheet included on the USB flash drive Mechanical stage tension is loose gt Tighten the stage brakes on the stage knobs as desired to increase tension Note For additional technical support contact your local EVOS distri
26. cope www amgmicro com EVOS User Guide REV C6 Published 4 JUN 2012 lt AMGS SETUP a DVANCED MICROS LED LIGHT CUBES IMPORTANT Before changing light channels ALWAYS be sure the light cube lock has been removed Applying force to the light cube selection lever while the lock is in place may seriously damage the mechanism This type of damage is not covered by the manufacturer s warranty 1 Move the stage back to allow access to the light cube lock which is centered under the back of the stage LED light cube lock 2 Loosen the thumbscrew to remove the light cube lock You may store the light cube lock in your accessories box for future use during transport or shipping 3 Place the light cube access cover into the opening and tighten thumbscrew Note For information about adding optional LED light cubes refer to Changing LED Light Cubes p 23 Secure light cube access cover over opening POWER SUPPLY 1 Turn the power switch to the O OFF position before connecting the power adapter IMPORTANT Always use the correct power supply The power adapter specifications appear on the serial number label front of LCD hinge and in the SPECIFICATIONS p 36 Damage due to an incompatible power adapter is not covered by warranty Power switch Power adapter plugged in 2 Connect the power adapter to the power jack on the right side of the microscope base attach the c
27. e dialog box gt Save File Name text field Click to enter a file name gt New Folder button Click to enter a folder name and date optional gt Save Folder list The captured image will save to the selected orange folder gt File list to the right of the Save Folder list Displays saved files already in the selected folder gt File type menu Drop down menu allows tif png jpg or bmp formats gt Comment text field Click to enter a comment and date optional Note For more information refer to Saving Images amp Working with Files p 14 VIRTUAL KEYBOARD The Virtual keyboard popup allows text entry anytime a text field or text related button is selected Where applicable a Date button is available to include the current date in the text field Click the Accept button when you have finished entering the text Note You may plug in a USB keyboard for text input When a keyboard Is connected the virtual keyboard does not pop up except when you add a new user ID or rename a file with the Image Review tool INFO DISPLAY monochrome camera version only The Info display bottom right shows the following context sensitive data in the Find amp Focus and Actual tabs gt Intensity Max and Min gt Ratio Max to Min gt Cursor position gt Current objective selected In Overlay the info display shows the QuickSave option with the QuickSave file name see QuickSave tab under Settings but
28. e slider usage guidelines p 77 Fluorescence Applications The Block slider is useful in a dark environment to block fluorescent light from being reflected by the condenser and improve image quality It allows more access to the sample than the light shield box Condenser slider slot MOUSE SCROLL WHEEL When the Enable Mouse Wheel option is selected in the Basic tab of the Settings dialog box see p 28 the mouse scroll wheel will adjust the illumination intensity Roll the scroll wheel away from you for brighter illumination or roll it toward you for dimmer illumination www amgmicro com EVOS User Guide REV C6 Published 4 JUN 2012 GENERAL CARE gt When cleaning optical elements use only optical grade materials to avoid scratching soft lens coatings gt Use the appropriate cleaning solutions for each component as indicated in the Sterilization Procedures below gt If liquid spills on the microscope turn off the power immediately and wipe dry gt Do not exchange objectives between microscopes unless you know that the components have been approved and recommended by AMG gt After using cover the microscope with the supplied dust cover IMPORTANT Never disassemble or service the microscope yourself Unauthorized repairs may damage the microscope or alter its functionality which may void your warranty Contact your local EVOS distributor to arrange for service OBJECTIVE LENS CARE Clean
29. e stage knobs condenser wheel etc Do not soak any surface in sterilization solution NEVER spray liquid anywhere on the EVOS Always wipe surfaces with dampened paper towels instead 4 If it is necessary to sterilize the condenser do not apply solution directly to the condenser assembly Instead select the desired phase ring and then cover the condenser with clear plastic wrap and wipe the wrap with sterilization solution IMPORTANT EVOS should not be subjected to UV sterilization UV degrades many materials including plastic Damage from UV exposure is not covered under the manufacturer s warranty www amgmicro com EVOS User Guide REV C6 Published 4 JUN 2012 IMAGE QUALITY ISSUES POSSIBLE SOLUTIONS PROBLEM Misaligned overlay image Transmitted light image is too dim at higher magnifications Specks dots or blurs on image Uneven focus across screen Trouble focusing on coverslipped specimen on standard slide LCD screen is black LCD screen is red or red patches cover parts of the screen gt Re capture images in each channel gt Set the phase annuli selector to the BF position gt Remove condenser slider if one is in place gt Remove light shield box if it is in place gt Follow instructions under Objective Lens Care p 32 to clean objectives gt Position specimen flat on the stage be sure the specimen s thickness is even gt Place the slide so the coverslip is facing up
30. ead lighting might have on your image Condenser shield removable www amgmicro com EVOS User Guide REV C6 Published 4 JUN 2012 ee le Lit ie y i h Pe e a ae EVOS installed in a cell culture hood LCD tilted back into transport position www amgmicro com INSTALLING EVOS IN A CELL CULTURE HOOD EVOS small footprint simple power connection and easily viewed display make it quick to install and convenient to use in a cell culture hood DIMENSIONS EVOS will fit in cell culture hoods that are at least 20 12 inches 520 mm deep If your cell culture hood is smaller it may be possible to turn the EVOS at a slight angle to fit ENGLISH METRIC DEPTH 18 5 in 47 0 cm WIDTH 14 0 in Gave tcl HEIGHT TRANSPORT 12 75 in 32 4 cm HEIGHT DISPLAY 2275in 57 8 cm WEIGHT 33 7 Ibs 15 3 kg INSTALLATION Note Refer also to the illustrations on p 4 for more details about moving EVOS 1 Secure the stage with the stage lock pin switch EVOS off and disconnect the power cord mouse and if connected keyboard 2 Tilt the LCD screen back until it is parallel with the tabletop 3 Lift the microscope by grasping it firmly with both hands under the support arm just behind the condenser 4 Gently place the microscope on a lab cart and transport it to the cell culture hood Note Verify that the hood sash Is raised
31. enough for the microscope to slide underneath approximately 14 or higher 5 Lift the microscope as before and move it into the hood 6 Tilt the LCD monitor upright 7 Remove the stage lock pin connect the power cord mouse and if desired keyboard and switch EVOS on EVOS User Guide REV C6 Published 4 JUN 2012 Ses QUICK REFERENCE DIAGRAMS Channel indicator bar TRANSMITTED Active channel highlighted Login button Control bar Control bar tabs e Find amp Focus e Actual e Overlay LIGHT ON OFF button Illumination slider Exposure time slider Image capture button Color option Scalebar toolbar options Settings control button Save image button 34S ec 7 Info display bar ee A apie a a Selected objective on PS lem o Se eee eee pce o O O00 00 The color camera version shows the QuickSave option instead of the info display bar Power switch Power input jack USB and DVI ports Coarse stage positioning knobs Stage X axis knob Stage Y axis knob Focusing knobs Objective selection wheel Light cube selection lever Phase annuli selector Condenser slider slot Note Refer to the CONTROLS GLOSSARY p 26 for more details about onscreen and mechanical controls www amgmicro com 8 EVOS User Guide REV C6 Published 4 JUN 2012 Power switch and data ports Light cube selection lever Objective selection wheel and focusing knobs Find
32. er the sample This is important for optimal image quality Note If your application requires access to the sample work in a dark room and use the Block slider to block light reflected from the condenser 9 Move the light cube selection lever to the desired fluorescence channel the channel indicator bar will highlight the selected light cube 10 Click the Find amp Focus tab 11 Click the LIGHT ON button to turn on fluorescence illumination WARNING UV LIGHT HAZARD When using the DAPI channel avoid exposure to beam and use protective shields NEVER look directly at light Adjust the focus as needed continued on next page EVOS User Guide REV C6 Published 4 JUN 2012 CALLS BASIC OPERATION ADVANCED MICROS z CAPTURE 1 Illumination slider and Capture button in the control bar i SETTINGS SAVE a Ble Color options shown for fl monochrome Save button DISPLAY Color VIEW Scalebar Toolbar imti Ma Malin Lontrast T ra A be A 75 Brightness Reset a 4 TE DAPI G A 20 v a 32 RFP 5 29 SANE FILE VIRTUAL KEYBOARD Caps Lock Cancel Virtual keyboard Find amp Focus Actual LIGHT 1 ms Actual tab www amgmicro com 13 14 15 16 17 18 19 20 Fluorescence Operation continued Adjust the illumination intensity if necessary using the Illumination slider on the control ba
33. ers on that server You may now navigate below the top level shared folders on the Network page After the server accepts your login EVOS will display the list of available shared folders on the selected server Select a shared folder and click the Add button to include it in the list of possible file destinations Alternatively you may type in the file path and click Add The folder should appear on the list box below the Add button If it does not contact your network administrator for help Note You may add multiple shared folders to the list but you can only use a single login on any one server or domain We recommend that each EVOS user establish network connections under his or her own EVOS user ID See Logging In Creating New User Logins p 13 for more details on EVOS user IDs Your EVOS user ID will remember your network login it is encrypted in the microscope s non volatile memory All your connections and login information will be hidden from other EVOS user profiles If you need to remove a shared folder from the destinations list select the folder name and click the Remove button Click OK to close the Settings dialog box To verify your list of network destination folders go to the QuickSave tab and click the Browse button to display the QuickSave Browse popup All your selected network destinations as well as any USB flash drives currently plugged in will appear in the Browse popup
34. g is missing Note If you do not have your distributor information you can look it up at the or contact AMG Customer Service see p 35 LED light cube lock LED light cubes in place under EVOS microscope per order in place under stage LED light cube lock Condenser shield removable Power adapter Dust cover USB flash drive includes User Guide and Quick Start Guide gt Accessories e Light shield box e USB mouse e Power cord e UV shield assembly mount shield screws amp L shaped hex key e Light cube access cover includes LED light cube installation tool install light cube access cover before using EVOS v v v v v CLASS 3B LED UV RADIATION Light cube access cover top Light cube access cover bottom with tool UV shield assembly kit Light shield box www amgmicro com EVOS User Guide REV C6 Published 4 JUN 2012 lt AMGS SETUP a DVANCED MICROS MOVING TRANSPORTING EVOS gt ALWAYS lock stage with the stage lock pin O before moving microscope gt When transporting or shipping EVOS secure the LED light cubes in place with the light cube lock gt Lift the microscope by grasping it firmly with both hands under the support arm balancing the weight as shown at left Stage lock pin engaged gt To transport EVOS to a different facility use the original packaging materials if possible Always be sure the microscope is properly cushioned and b
35. gaged only save at 8 bit depth 5 Click in the Comment text field to enter a comment and date optional 6 Click the Save button to save the file 7 To delete a file or folder highlight the item on the list and click the Delete button A confirmation dialog box will pop up It is not possible to recover a deleted file 8 To rename a file or folder highlight the item on the list and click the Rename button The virtual keyboard will pop up you can use the Clear button to reset to a blank field EVOS User Guide REV C6 Published 4 JUN 2012 zAV C gt ADVANCED OPERATION ADVANCED MICROSCOPY GROUP F VIEW SETTINGS SAVE intensity INFO Scalebar ee Max 163 Ratio 24 0 40x Toolbar Min 6 Cursor 12 Objective Settings button and Save button SETTINGS Basic Quick Save Channels Network Service OuickSave File Base Filename Fluor Count 1 File Type lif v Next Fluor _RGB_0001 10 DuidkSawe Folder Browse lt O USBI Also save each channel separately 16 bit 10 Cancel Settings QuickSave Tab VIRTUAL KEYBOARD xT New Folder 04 01 2010 Backspace Clear 9 lt lt Date Caps Lock Accept Cancel Virtual keyboard BROWSE Folder New Folder Vv USBI gt DOM Experiment 1 FL new Ul captures OK Cancel Browse popup to select a destination folder OUICKSAVE Fluor RGB 0001 tif 40x Objective QuickSave opti
36. his will create up to five files per captured image named according to the following conventions gt BaseName_RGB_0001 tif Overlay image gt BaseName_channel_0001 tif where channel is the selected channel such as GFP RFP DAPI or TRANS When optional LED light cubes are installed the files will automatically save channels with their names See Changing LED Light Cubes p 23 for more information 8 Click OK to accept QuickSave settings 9 Select the Overlay tab and click the radio button for the QuickSave option Note The color camera version displays the QuickSave option in all tabs 10 After acquiring an image with the Capture button click Save The image will be saved as specified in the QuickSave settings EVOS User Guide REV C6 Published 4 JUN 2012 lt 5 ADVANCED OPERATION ADVANCED MICROSC X axis and Y axis stage brakes VIEW SETTINGS Toolbar bat TOOLBAR Y Time Lapse _ Start Settings 5 Value Unit Interval 1 Minute Toolbar option in the control bar TOOLBAR P Time Lapse P Transfection Duration 30 Minute r images Oo a1 Folder 7 Browse an 8 USBI1 P Count P image Review Toolbar Base Filename Cells File Type 10 eas tif v First Cells 0001 0f Write Video File Cells avi Time Lapse tool TIME LAPSE PROGRESS Interval 30 Seconds Remaining Seconds 18 Duration 5 Minutes
37. long working distance objectives require a thick optical substrate and image best through 1 0 1 5 mm of glass or plastic gt Click the LIGHT ON button onscreen gt Move objective selection wheel so that light shines through objective gt Verify that the phase annuli selector on condenser is not stuck between settings gt Center specimen over objective gt Verify power supply is connected and power switch is on gt Dim the illumination until the red highlights disappear to get the maximum level of brightness without any overexposed areas gt Disable the Highlight saturated pixels in red option in the Settings see p 28 SOFTWARE INTERFACE ISSUES We recommend keeping your EVOS microscope up to date with the latest software See the Updating Software section for more information PROBLEM Image does not respond to changes in focus or stage position LIGHT ON OFF button is inactive Scalebar does not appear when clicked Save button does not respond when clicked DVI output does not work on an external LCD monitor or projector Unable to connect to network www amgmicro com POSSIBLE SOLUTIONS gt Click the LIGHT ON button note that a red USB icon on the Save button indicates there is an unsaved image which will be lost unless it is saved before clicking LIGHT ON gt Verify that the light cube selection lever is clicked into a position gt Verify that the objective selection wheel is cli
38. lor Actual On 1 1 Seg dh 30ms Actual tab Brightness Reset Contrast TRANS A TE 4 Fa Find amp Focus tele w _ if 21 Ne A 789 Actual aly a 32 A 53 Overlay RFP v 29e 75 Overlay tab DISPL AY ADJUST COLOR Brightness A Color meee 7 Saturation A Color Adjustment Hue k button zz 3 Reset Cancel Adjust Color Dialog O a Scalebar VIEW ih scalebar ay Teolbar View Scalebar and Toolbar options TOOLBAR P Time Lapse b Transfection P Count P image Review Toolbar www amgmicro com ACTUAL TAB Use the Actual tab with fluorescence channels to view the image at the actual exposure time used for high quality image capture With Actual tab selected EVOS responds more slowly to stage position and focus changes depending on the user selected exposure time for the camera The following controls are available in the Actual tab LIGHT ON OFF button Illumination slider gt gt gt Exposure time slider gt Image capture button gt Color option or Color Adjustment Button OVERLAY TAB Use the Overlay tab to select and overlay multiple fluorescence channels as a single multicolor image The following controls are available in the Overlay tab gt Brightness and Contrast sliders for each channel gt QuickSave option to the right not shown ADJUST COLOR DIALOG BOX The Color Adjustment button left control bar opens the Adjust Color dialog
39. m or transmitted in any form or by any means for reasons other than personal use without the express written consent of AMG Information in this document is provided in connection with AMG products No license express or implied by estoppel or otherwise to any intellectual property rights is granted by this document www amgmicro com EVOS User Guide REV C6 Published 4 JUN 2012 Optics Infinity corrected optical system RMS threaded objectives with 45 mm parfocal distance Objectives included vary per order Contact AMG Customer Service see p 35 or contact info amgmicro com for a full list of objectives supported Objective Turret 5 position front mounted control Light cubes included vary per order Specifications for our most popular light cubes are listed below Contact AMG Customer Service for a full list of LED light cubes supported Objectives Light Cubes gt U S Patent No 7502 164 DAPI 360 nm excitation 447 nm emission GFP 470 nm excitation 525 nm emission RFP 530 nm excitation 593 nm emission illumination LED 50 000 hour life adjustable intensity Contrast Methods Fluorescence and transmitted light brightfield amp phase contrast Includes 3 pos with brightfield and phase contrast annuli Condenser ora Working Distance X Y axis fine positioning controls 28 5mm 1 11 per rotation Mechanical 110 mm x 110 mm 4 3 x 4 3 range of motion Glide Stage Z axis f
40. mizes photobleaching and phototoxicity effects When you capture an image illumination and exposure time automatically adjust for best image quality and then reset to lower levels after the capture The Actual tab provides full powered illumination and actual exposure times for live viewing of the sample Note With longer exposure times more than 200 ms there will be a lag between focusing the image and seeing the focus change onscreen EVOS User Guide REV C6 Published 4 JUN 2012 zAV C gt BASIC OPERATION WV NN GDN EINO s BRIGHTFIELD OR PHASE CONTRAST OPERATION 1 Turn on the microscope using the power switch on the right side of the microscope base 2 Plug a USB flash drive into one of the USB ports on the right side of the microscope arm Power switch and data ports Light cube selection lever j 3 Place the sample on the stage using a vessel holder if needed 4 Set the magnification using the objective selection wheel on the front of the microscope 5 Move the light cube selection lever on the left side of the microscope all the way toward you the channel indicator bar will highlight the Transmitted position 6 Turn the phase annuli selector to the position that corresponds to the selected objective and contrast method 7 Insert the appropriate condenser slider into the slot on the condenser assembly for optimal image quality Note A Diffuser slider can
41. nging settings in the Service tab will affect the microscope s performance If service beyond the date time setting is needed please contact your EVOS distributor 2 Click the Change button under Date and Time in the Service tab 3 Inthe Date and Time popup use the arrows and the drop down menus to set the time date and time zone 4 Click OK in the Date and Time popup 5 Click OK in the Settings dialog box The new settings will take effect immediately EVOS User Guide REV C6 Published 4 JUN 2012 Channel indicator bar with GFP channel highlighted Guest Control bar variations Find amp Focus Actual amp Overlay Note In all tabs the color camera version displays the QuickSave option instead of the Info bar CAPTURE i DISPLAY Actual E Color Find amp Focus tab DISPLAY Color Color Adjustment button www amgmicro com This section describes all the onscreen and mechanical controls in detail To see the most commonly used controls in context refer to the QUICK REFERENCE DIAGRAMS p 8 ONSCREEN CONTROLS This glossary is not alphabetized Onscreen items are listed from top to bottom first and then from left to right CHANNEL INDICATOR BAR The channel indicator bar top highlights whichever one of the following light channels is currently selected gt Lever position 1 transmitted in example closest to front of microscope gt Lever
42. nsions 3 If the image or video file you wish to review is not in the directory displayed click the Browse button to find and open the desired directory 4 Use the scroll bar as needed to search the preview list for the desired file Click the image to select it The selected file name appears orange 5 Click the View File button to display the image in the image review window This button toggles between View File and Hide File Hiding the file closes the image review window Note Double clicking the thumbnail image will also toggle between displaying and hiding the file 6 To zoom the image in the review window double click the area of interest right click to restore normal magnification Refer to Using Digital Zoom p 20 for more detailed instructions 7 To rename a file select it and click the Rename button The Virtual Keyboard will pop up Enter the new file name and click the Accept button 8 To delete a file select it and click the Delete button A confirmation dialog box will pop up It is not possible to recover a deleted file 9 Use the Slide Show button to display each image thumbnail within the image review window continuously EVOS User Guide REV C6 Published 4 JUN 2012 zAV C gt ADVANCED OPERATION DVANCED MICROSCOPY GROUP Live image at 40x magnification Live 40x image zoomed 2x gt Oe vr a Pee sued k E X lt 4 L gt A AJ p
43. nter the new name CHANGE THE DEFAULT LOGIN The default user login is Guest to set the default login as the last active user go to the Basic tab of the Settings dialog box and uncheck the Default to Guest on startup option Note For multiple users we recommend leaving the Default to Guest on startup option checked EVOS User Guide REV C6 Published 4 JUN 2012 lt 5 ADVANCED OPERATION ADVANCED MICROSC DISPLAY ie SETTINGS SAVE intensity INFO Scolor Scalebar Max 168 Ratio 24 0 tt a Min 6 Toobar Cursor 12 Save button O Scalebar option and Display options SAVE RILE Homensary chew ice auai la bike flease inserta ush tach owe Save File information message SAVE FILE 4x Hela Cells Confluent tif image 0001 Save File dialog box VIRTUAL KEYBOARD x New Folder 04 01 2010 Backspace 17 Clear O gt lt e Date Shift K C Caps Lock o Accept Cancel Virtual keyboard CONFIRM FOLDER DELETE Please confirm that you wish to pemmanently delete the Experiment 1 folder Yes No Confirmation popup www amgmicro com SAVING IMAGES amp WORKING WITH FILES When you click the Save button the Save File dialog box appears If there is no USB flash drive or network connection in place an information message will appear Click the Cancel button to clear this message In the Save Folder list and the saved files list selec
44. ocusing controls 480 um rotation Interchangeable vessel holders available for most common shapes amp sizes LCD Display 15 color 1024 x 768 pixels adjustable tilt images f Onboard microprocessor built in software for image acquisition via mouse control Acquisition Monochrome Color Camera Sony ICX445 monochrome CCD Sony ICX285AQ color CCD 1 3 1280 x 960 1 3 Megapixels 2 3 1360 x 1024 1 4 Megapixels Monochrome camera Color camera Captured 16 bit monochrome TIFF or PNG 12 bit dynamic 16 bit color TIFF or PNG 12 bit dynamic range Images range 24 bit color TIFF or PNG 24 bit color TIFF or PNG joeg bmp 1280 x 960 pixels joeg bmp 1360 x 1024 pixels Output Ports 3 USB and 1 DVI digital output Power Suppl AC Adapter Inout 100 240V 50 60Hz sad Output 12 VDC 415A OCO Operating height 57 8 cm 22 75 Storage transport height 32 4 cm 12 75 Depth 47 0 cm 18 5 Width 35 5 cm 14 0 Note Specifications are subject to change without notice Go to the EVOS product page online to download the latest product information www amgmicro com EVOS User Guide REV C6 Published 4 JUN 2012 AMG O E www amgmicro com info amgmicro com 866 614 4022 01 425 368 0444 outside the US
45. on radio button www amgmicro com USING THE QUICKSAVE OPTION QuickSave allows you to save multiple images under a single base file name Simply specify the settings and select the QuickSave option and EVOS will save each image with a single click of the Save button 1 Click the Settings button to open the Settings dialog box and then select the QuickSave tab 2 Click in the Base Filename text field and enter a name that describes the imaging session The orange Next file name will reflect the information entered 3 Click in the Count text field to enter the starting number if you do not want to start at 1 The orange Next file name will reflect the information entered 4 Selecta file format tif or png from the File Type drop down menu 8 5 Click the Browse button to select a destination folder for the QuickSave files In the Browse popup highlight the desired folder and click OK 6 To create a new folder first click the name of the desired parent folder and then click the New Folder button 8 Enter a folder name and date if desired After creating the new folder click OK to close the Browse popup Note Clicking the Date button anywhere within a text field will automatically insert the current date MM DD YYYY wherever the cursor Is in that field 7 Select Also save each channel separately to save multiple channels for each image within an overlay image T
46. ool under it 4 Move the light cube selection lever to the position you want to use for the new cube 5 Use the light cube tool to loosen the 2 slotted screws as shown in photo at left 6 Screw the threaded end of the light cube tool into the hole in the center of the light cube as shown 7 Use the tool to tilt the light cube slightly toward you and lift out gently and then remove tool from cube 8 Attach the tool to the new light cube and then lower the cube into position so that the electronic connection aligns properly facing the back of the microscope and the cube sits squarely in place 9 Use the light cube tool to gently tighten the 2 slotted screws O so that the screw heads sit flush with the ridges on the light cube 10 Slide the tool into its storage slot 11 Replace the light cube access cover and ensure the thumbscrew is tightened 12 Turn the power switch to the ON position ASSIGN CUSTOM CHANNEL DISPLAY COLORS You may assign custom display colors for each light cube in the Channels tab of the Settings dialog box Note The color camera version does not include this feature 1 Click the Settings button to open the Settings dialog box and then select the Channels tab 2 Select the appropriate color from the drop down menu for each light cube you wish to customize 3 Click OK to accept the custom color assignments The display change will take effect with the
47. ord to the adapter and plug the cord into an outlet USB PORTS Plug the mouse and the USB flash drive into the USB ports located on the bottom right of the support arm You may also plug in a USB keyboard not included for text input DVI OUTPUT PORT A DVI port is available for output to a projector or other display DVI cable not included This port produces digital output only EVOS is compatible with either a DVI D or DVI I display Mouse and USB flash drive DVI cable not included plugged in plugged in www amgmicro com EVOS User Guide REV C6 Published 4 JUN 2012 UV SHIELD A WARNING UV LIGHT HAZARD This microscope uses a Class 3B ultraviolet LED for the DAPI channel Avoid exposure to the UV beam and use protective shields NEVER look directly at light For your protection follow this procedure to install the UV safety shield before using the DAPI fluorescence channel 1 Attach the UV shield mount to the front of the condenser with the two screws provided 2 Remove the protective coverings from both surfaces of the UV shield 3 Place the holes in the UV shield over the screws on the mount and push the slots down on the screws to secure the shield in place as shown Note The UV shield is removable for access to the condenser sliders used in transmitted light mode Simply unhook it from the screws on the UV mount CONDENSER SHIELD This shield can help reduce the effects that overh
48. position 2 GFP in example second from front gt Lever position 3 RFP in example second from back gt Lever position 4 DAPI in example closest to back See also Light Cube Selection Lever p 31 LOGIN BUTTON The Login button AMG logo bottom left allows for logging in and creating or changing user profiles This button also displays the current user profile CONTROL BAR The control bar bottom left varies depending on which tab is selected Choose a tab by clicking on it in the left end of the control bar gt Find amp Focus to avoid photobleaching while setting up a fluorescence specimen gt Actual to view image using actual acquisition parameters LED brightness and exposure time gt Overlay to view multiple fluorescence channels also the QuickSave option is available on this tab FIND amp FOCUS TAB Use the Find amp Focus tab to view the sample with either transmitted light to minimize photobleaching or fluorescence and to acquire images This feature displays 10 frames second for focusing and captures images at longer exposure high quality settings The following controls are available in the Find amp Focus tab gt LIGHT ON OFF button gt Illumination slider gt Image capture button gt Color option or Color Adjustment Button EVOS User Guide REV C6 Published 4 JUN 2012 lt 5 CONTROLS GLOSSARY ADVANCED MICROSC Find amp Focus LIGHT CAPTURE DISPLAY Co
49. r or the mouse scroll wheel Note When the Color option is off overexposed pixels will appear red Dim the illumination until the red highlights disappear to get the maximum level of brightness without any overexposed areas See p 28 for instructions on changing the overexposed pixel display For the color camera version the overexposed pixels are always highlighted in red unless this feature is disabled in the settings Click the Capture button to acquire the image Move the light cube selection lever to the next position and repeat steps 10 14 to acquire each fluorescence channel as desired Click the Overlay tab to show all channels in color overlay Adjust brightness and contrast for each channel to bring them into balance with each other Click the Save button to save the color image The Save File dialog box will pop up Click in the Save File Name text field to enter the file name A virtual keyboard will pop up After entering the file name click the Accept button at the lower right of the keyboard Choose the file type and click the Save button Note See Saving Images amp Working with Files p 14 for more information HELPFUL TIPS b Turn off the Color option to display the image in grayscale This often shows more detail than a color image Find amp Focus uses a shorter exposure time 100 ms and lower illumination appx 60 compared to image capture settings This mini
50. raced to prevent damage Gi IMPORTANT Never allow EVOS to be subjected to sudden impact or excessive vibration Handle the microscope with care to prevent damage Grasp under support arm with both hands to lift EVOS OPERATING ENVIRONMENT gt Place the microscope on a level surface away from vibrations from other pieces of equipment gt Allow at least 5 cm 2 in free space at the back of the microscope to allow for proper ventilation and prevent overheating of electronic components gt Set up EVOS away from direct light sources such as windows Ambient room lighting can enter the imaging path and affect the image Note Place the light shield box on the stage over the sample to reduce the effects of ambient light and improve image quality Operating temperature range 4 32 C 40 90 F gt Relative humidity range 30 90 IMPORTANT EVOS should not be subjected to UV sterilization UV degrades many materials including plastic Damage from UV exposure is not covered under the manufacturer s warranty Light shield box on stage MECHANICAL STAGE STAGE LOCK PIN Before moving the mechanical stage for the first time remove the stage lock pin from the back right hand corner of the stage plate Pull firmly to remove this pin You may store the stage lock pin in your accessories box for future use Note Always secure the mechanical stage with the stage lock pin before moving the micros
51. ted items will appear orange If a USB keyboard is installed the virtual keyboard will not appear Note that pressing the Enter key on a physical keyboard is like pressing the Save button in the Save file dialog box Note Saved image files include the date time stamp To ensure accurate date time information verify the settings before your capture session See Setting Date and Time p 25 for instructions 1 Click in the Save File Name text field Enter a file name and click the Accept button To overwrite a file simply select the name of the file from the saved files list instead of clicking on the Save File Name text field A Save As confirmation dialog box will pop up It is not possible to recover an overwritten file 2 Click on the name of a folder in the Save Folder list to select the destination for the new image 3 To create anew folder first click the name of the parent folder and then click the New Folder button to enter a folder name and if desired date Note Clicking the Date button anywhere within a text field will automatically insert the current date MM DD YYYY wherever the cursor is in that field 4 Select a file format tif ong jog or bmp from the File Type drop down menu Note To save a 16 bit image select tif or png and ensure the Scalebar and Color options are off File types jog and bmp as well as images of all types with the Scalebar or Color options en
52. ton p 28 and the current objective selected DIGITAL ZOOM VALUE DISPLAY The Digital Zoom Value display bottom right appears above the selected objective display to show the zoom value when you zoom an image EVOS User Guide REV C6 Published 4 JUN 2012 O gt CONTROLS GLOSSARY DVANCED MICROSCOPY GROUP Coarse stage positioning knobs stage X axis knob brake and stage Y axis knob brake Focusing knobs and objective selection wheel www amgmicro com MECHANICAL CONTROLS This controls glossary is not alphabetized Mechanical controls are listed in the order they are normally used POWER INPUT JACK Plug the power adapter into the power input jack O POWER SWITCH Set the power switch to to turn the microscope on or O to turn it off USB AND DVI PORTS Plug the mouse and flash drive into the USB ports Use the DVI port for digital output to a projector or other display COARSE STAGE POSITIONING KNOBS Use the coarse stage positioning knobs to position the specimen within the field of view particularly at low magnifications STAGE X AXIS KNOB Use the stage X axis knob for fine left right movements to position the specimen within the field of view particularly at high magnifications To secure the stage in place for time lapse image captures tighten the X axis stage brake STAGE Y AXIS KNOB Use the stage Y axis knob for fine front
53. verexposed pixel display Click the Capture button to acquire the image Click the Save button to save the image The Save File dialog box will pop up Click in the Save File Name text field to enter the file name A virtual keyboard will pop up After entering the file name click the Accept button at the lower right of the keyboard Choose the file type and click the Save button Note See Saving Images amp Working with Files p 14 for more information EVOS User Guide REV C6 Published 4 JUN 2012 Guest Find amp Focus LIGHT A kj Actual AMG Login button set to Guest profile Al Users Add 0 Copy from Larry ae 4 Remove lt Rename ae 7 Kn lt Cancel Login dialog box VIRTUAL KEYBOARD New Folder Caps Lock Cancel Virtual keyboard SETTINGS Basic Quick Save Channels Network Service This EVOS F1209 1441 026 Software Revision 15844 Serial Number Login Default to Guest on startup Display Highlight saturated pixels in red Reset Scalebar Location Enable Mouse Wheel Mouse Speed LCD Backlight Less LCD Backlight More File Save Always save TIFF as 8 bit Save TIFF in Microsoft compatible format OK Cancel Settings Basic Tab to change default login www amgmicro com LOGGING IN CREATING NEW USER LOGINS EVOS keeps settings in memory for each user ID so multiple users c
54. z 4A 4 19 nP O a WF T Tk g x Prey f t h S W F aptured image at 40x magnification with scalebar www amgmicro com USING DIGITAL ZOOM EVOS can zoom the image onscreen quickly allowing a closer look Simply double click live or captured images to zoom them In the images below the numbered arrows indicate click points Also note that the zoom factor display appears over the selected objective display ZOOM AND RECENTER LIVE IMAGES Note Live images can only zoom to 2x To zoom an image at higher levels you must first capture it 1 Double click the area of interest O in the image onscreen EVOS will display a view zoomed 2x centered on the point clicked 2 Inthe enlarged image double click on any point in the middle area of the screen to recenter the image EVOS will place points from the outer edges of the screen as close to the center as possible You may recenter repeatedly 3 Right click anywhere on the image to restore the view to unzoomed magnification ZOOM CAPTURED IMAGES 1 Double click the area of interest in the image onscreen A view zoomed 2x centered on the point clicked will appear 2 Inthe enlarged image double click again on any point to double the digital zoom level The enlarged image will center around the point clicked 3 Continue double clicking to double the digital zoom value as desired It is possible to zoom in to the pixel level of the

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