Quick Guide
Contents
1. Adjusting the laser intensity e Set the Pinhole to 1 Airy Unit Fig 23 e Set the Detector Gain high Fig 22 Image Display e When the image is saturated reduce AOTF transmission in the Laser control section of the Channels Tool Fig 23 using the slider to reduce the intensity of the laser light to the specimen Adjusting gain and offset w Channels Channel Ch2 Ch3 e Increase the Amplifier Offset until all blue pixels disappear and then make it slightly positive Fig 23 Pinhole 1 00 Airy Units 116 pm section e Reduce the Detector Gain until the red pixels only just disappear Gain Amp Offset Track Tracki v v 488 514 561 633 9800 Transmission Fig 23 Channels tool 18 08 2007 Scanning a Z stack ZStack Open the Z Stack tool in the Left Tool Area Select Mode First Last on the top of the Z Stack tool Continuous Click on the button in the Action Button area A continuous XY scan of the set focus position will be performed Use the focus drive of the microscope to focus on the upper position of the specimen area where the Z Stack is to start Click on the Set First button to set the upper position of the Z Stack Then focus on the lower specimen area where the recording of the Z Stack is to end Click on the Set Last button to set this lower position Optimal Interval 3 27 ym Click on the button to set number of slices to match the optimal2. Click the Laser icon to select the laser lines and set the attenuation values transmission in in the displayed window For the configuration of the beam path please refer to the application specitic configurations depending on the used dyes and markers and the existing instrument configuration In the Imaging Setup tool the Detection Bands amp Laser Lines are displayed in a spectral panel Fig 16 to visualize the activated laser lines for excitation vertical lines and activated detection channels colored horizontal bars 08 2007 Settings for track configuration in Channel Mode w m Imaging Setup Configuration Mode Channel Mode Switch track every Frame A Trakl ch 500 600 700 Fig 15 Imaging Setup tool for a single track LSM Activation deactivation of the excitation wavelengths check box and setting of excitation intensities slider If necessary open the Laser Control tool see above Selection of the main dichroic beam splitter HFT or secondary dichroic beam splitter NFT position through selection from the relevant list box Selection of an emission filter through selection from the relevant list box Activation deactivation via check box of the selected channel Ch 1 4 monitor diode ChM META detectors ChS1 8 transmission ChD for the scanning procedure and Fig 16 Detection Bands amp Laser Lines display EE imaging Seulp e For storing a new configuration enter a
3. Configuration IT cre m desired name in the first line of the Mode a a EE Configurations list box Fig 17 and click Alexa 488 Cy3 sim Switch track every duo seq Store NLO 1 a i i l T au EE e For loading an existing configuration select it Ch ose 1 a eee from the list box and click on the Load test meta b Utton z stack config e For deleting an existing configuration select it in the list box and click on Delete Fig 17 Track Configurations window Settings for multiple track configurations in Channel Mode Multiple track set ups for sequential scanning can be defined as one configuration Channel Mode Configuration to be stored under any name reloaded or deleted The maximum of four tracks with up to 8 channels can be defined simultaneously and then scanned one after the other Each track is a separate unit and can be configured independently from the other tracks with regard to channels Acousto Optical Tunable Filters AOTF emission filters and dichroic beam splitters The following functions are available in the List of Tracks panel in the Imaging Setup Tool Fig 15 Fig 16 and Fig 17 Switch track every Line Tracks are switched during scanning line by line The following settings can be changed between tracks Laser line laser intensity and channels Frame Tracks are switched during scanning frame by frame The following settings can be changed between tracks Laser line and intensity a
4. Z interval for the given stack size objective lens and the pinhole diameter Make sure that the activation tickbox is set gt Click on the Start button to start the recording of the Z Stack Mode First Last Center Number of Slices Interval Keep Interval Slice Optimal Interval 3 08 ym Set Last 20 241 Set First 25 695 Range 45 94 um Focus Position 2 73 ym Z Stack Navigation Move to Last Center First Fig 24 Z Stack tool Note When ETOD is not activated the respective tool and its set parameters are not included in the multidimensional image acquisition If no multidimensional tool is activated the pan Start button is grayed out and only single images can be scanned 08 2007 S Fig 25 Save Image buttons in ZEN speichern unter Speichern ej Desktop IE Eigene Dateien demo data Fi Arbeitsplatz 2 Jenakultur ONetzwerkumgebung JofBioTech_CallforPapers gt CiscoSecure Authentication Agent 95m 4 2 Icons pix Movies Movies O 4xiovision 4 6 Icons Music x iT Dateiname Image2 Dateityp Fig 26 Save as window Tagged Image File Contents of image window single plane Raw data single plane Raw data series Contents of image window single plane Contents of image window series Full resolution image window single plane Full resolution image window series Select file name and save Fig 27 Export window 20 t
5. and the System PC switch Fig 1 e Switch off the X Cite 120 lamp or the HBO 100 mercury burner e Switch off the UV Ar laser of by the toggle switch on the power supply Fig 2 08 2007 21
6. bit 48 kB 0 44 MB 08 2007 Turning on the lasers e Open the Laser Tool in the left tool area always the first in the list and activate the lasers you need for your experiment Fig 10 Remember the argon multi line laser has to first be put to standby for a 5 minute warm up before it changes to on e Zeiss recommends operating the Argon multi line Laser at a tube current of about 6 A 50 output This is the best compromise between laser stability oower and laser life time tube current control can be found in Pro Mode DPSS 561 10 561 Chameleon 720 930 HeNe633 633 Continuous Laser Informaton Setup Manager Laser OD Imaging Setup O E Light Path Maximum Power 30 0 mW Wavelength 456 477 466 514 nm Online Acquisition Acquisition Mode Status Warming up Multidimensional Acquisition Tu b E C urent 6 D A OD ZStack Bleaching 4 Time Series Q Tile Scan i Information On Experiment Fig 10 Laser Control tool in Pro Mode Output 08 2007 Setting up the microscope Changing between direct observation camera detection and laser scanning mode The Ocular Camera and LSM buttons switch the beam path and indicate which beam path is currently in use for the microscope e Click on the Ocular button to change the microscope beam path for direct observation via the eyepieces of the binocular tube lasers are blocked e Click on the Camera button to change
7. Another unique feature in Imaging Software is the scalable ZEN interface This Workspace Zoom allows adjustment of the ZEN window size and fonts to the situational needs or your personal preferences Fig 7 e Setting up conventional confocal software for a specific experiment can take a long time and is often tedious to repeat With ZEN these adjustments have to be done only once and may be restored with just two clicks of the mouse For each type of experiment one can now set up and save the suitable Workspace Layout These configurations can also be shared between users e For most controls buttons and sliders a tool tip is available When the mouse pointer is kept over the button a small pop up window will display which function is covered by this tool button These are just some of the most important features of the ZEN interface For a more detailed description of the functionality for the ZEN software please refer to the User Manual that is provided with your system 6 08 2007 To create a new image document in an empty image container click the Single or the Start L button The new document is immediately presented in the Open Images Area Remember an unsaved 2D image in the active image tab will be over written by a new scan Multi dimensional scans or saved images will never be over written and a new scan will then automatically create a new image document Alternatively you can create a new empty image docu
8. Microscopy from Carl Zeiss LSM 5 MP LSM 510 and LSM 510 META LSM Software ZEN 2007 We make it visible Page NS ta tr rete cee erat es ee a eee EEEE 1 PVCU OCCU Oooo sasistese se resesesesntevaccuptocesnsesecneosetecsessedosespecSeuaeesetasqonseSesseesqSeunenseteseaesedeusnessasnecteeee 1 Starting the System casas access oceans eee tesco sinee se anccan son suee tansesesuancuseseonsueettanseeeseree 2 Introduction to ZEN Efficient Navigation ccccesseeeeseeeeeeeeseeeeeeseeeseaseeeeeasaeeseaneessaaaes 5 Setting up the TI ICl OS CO Care siactasececespanmaneneceaancnineceaeucuanesenseenaneusannecaneeeasucasmensdansuanasenseecaars 10 Configuring the beam path and laserS cceseceeeeeeeeeeeeeeeeeseaseeeseaaeeeeeaaneessaaseeseaneessaaaes 12 Scanning an NIT A Soe coc ates sie cca e cle notoriteceaecounecowetiocatiencsinaocareciewndonsiieonaineceume come ennnen mnnn ennnen mnnn 15 Storing and exporting image data cccccseeeeeeeeeeeeseeeseneeeseneeeceseeeeeneeseaneesoanessennessaneessaaes 20 Switching off the SV SCI sseccceesteetec eter et cecties tee ediesseqedincteentecccientensiewedcendanstountnestenetcecdavet cate 21 Introduction This LSM 510 LSM 510 META LSM 510 NLO Quick Guide describes the basic operation of the LSM 510 LSM 510 META LSM 510 NLO Laser Scanning microscope with the ZEN software The purpose of this document is to guide the user to get started with the system as quick as possible in o
9. ent S LS LS LS LS Fig 19 Channels tool 16 08 2007 Image acquisition Once you have set up your parameter as defined in the above section you can acquire a frame image of your specimen Use one of the Find Fast Continuous or Single buttons to start the scanning procedure to acquire an image Scanned shown in windows images are separate Click on the Stop button to stop the current scan procedure if necessary nd Select Find for automatic pre adjustment of detector gain and offset ast Select Fast for continuous fast scanning useful for finding and changing the focus Select Continuous for continuous scanning with the selected scan speed Continuous Select Single for recording a single image Select Stop for stopping the current scan procedure Image optimization Choosing Range Indicator In the View Dimensions View Option Control Block click inside the color field in the button under the channel button Fig 21 Note Clicking on the right hand side of the button leads to a list of colors 08 2007 View Dimensions Z Position 4 Channel s Merged Chi Chi Fig 21 View Dimensions Control Block 17 The scanned image appears in a false color presentation Fig 22 If the image is too bright it appears red on the screen Red saturation maximum If the image is not bright enough it appears blue on the screen Blue zero minimum
10. ew Controls Fig 5 ZEN Main Application Window after Startup with several images loaded 08 2007 Introduction to ZEN Efficient Navigation ZEN Efficient Navigation is the new software for the LSM 5 Family from Carl Zeiss With the launch of this software in 2007 Carl Zeiss sets new standards in application friendly software for Laser Scanning Microscopy The ZEN Interface is clearly structured and follows the typical workflow of the experiments performed with confocal microscopy systems On the Left Tool Area Fig 4 D the user finds the tools for image acquisition image processing and system maintenance easily accessible via 3 Main Tabs Fig 5 1 All functions needed to control the microscope and to acquire images are bundled in the Acquisition Tools Fig 5 3 and 4 Arranged from top to bottom they follow the logic of the experimental workflow The area for viewing and interacting with images is centered in the middle of the Main Application Window the Center Screen Area Each displayed image can be displayed and or analyzed with many view options available through view tabs which can be found on the left side of the image According to the chosen view tab the required view controls appear in View Control Blocks below each image File management and data handling tools are found in the Right Tool Area see Fig 4 and Fig 5 Color and brightness of the interface have been carefully adjusted to the typical light conditio
11. for reflected light illumination via the power supply as described in the respective operating manual Switching on the Enterprise UV Ar Laser e If the UV laser is required switch it on via the toggle switch Fig 2 1 on the power supply It will be ready for operation after a few seconds Fig 2 Power supply of UV Ar laser 2 08 2007 Starting the ZEN software F e Double click the ZEN icon on the WINDOWS desktop to start the Carl Zeiss LSM wer software ZEN 2007 The ZEN Main Application Window and the LSM 510 Startup window appear on the screen Fig 3 Login ZEN 2007 Laser Scanning Microscope LSM 510 O Boot Status z m Start System Image Processing Login ZEN 2007 Laser Scanning Microscope LSM 510 Boot Status CP DIOs CP Video Switches CP Cameras CP HRZ CP Focus Fe ec G a The ZEN 2007 Main Application Window ES P CP Focus Start System Offlime Demo c LSM 510 Startup Window Fig 3 ZEN Main Application Window at Startup a and the LSM 510 Startup Window b and c In the small startup window choose either to start the system online Start System hardware for acquiring new images or in Image Processing mode to edit already existing images Toggle the little g symbol to view the Boot Status display and get the additional Offline Demo button option Choosing Start System initializes the whole microscope system and activates the entire sof
12. g the stage in X and Y using the XY stage Tine motion control 10 08 2007 Setting the microscope for reflected light e Click on the Reflected Light icon to open the X Cite 120 Controls and turn it on e Click on the Reflected Light shutter to open the shutter of the X Cite 120 lamp HBO100 e Click on the Reflector button and select the desired filter set by clicking on it Storing the microscope settings Microscope settings can be stored as configurations Fig 13 by typing a config name in the pull down selector and pressing the save button Fast restoration of a saved config is achieved by selecting the config from the pull down list and pressing the ka load button The current config can be deleted by pressing the delete button In Pro Mode these configurations can be assigned to buttons that are easier to press Note Depending on the microscope configu ration settings must be done manually if necessary 08 2007 amp E Light Path Reflected Light ore Source X Cite 120 Configuration or HBO 100 0 La Bem Reflected Light m wue Closed 100 63 6 Shutter a aam E Reflector kka Transmitted Light pea Control BF kd p pg 49 61 Fig 12 Microscope Control window with Transmitted Light pop up menu w E Light Path Ocular Fig 13 Saving Microscope configurations Configuring the beam path and lasers e Click on the LSM button Choosing a configuration Simultaneous
13. gs Administrator All Users Default User HPPET M105E Application Data amp Desktop Axiovision 4 6 Icons amp demo data 4D cells fromChris amp Nice Small LSM D Gif Movies Imaaes File Browser i f Alzheimers4D 3 Channels 8 bit 0 14 MB 0 60 MB cS 3 Channels 8 bit 0 29 MB 0 52 MB fons gt oe We es cervicalsmear 3 Channels 8 bit 0 13 MB 0 58 MB a Ke ce 3rd 3 Channels 8 bit 0 19 MB 0 75 MB Arabidopsisleats 3 Channels 8 bit 75 kB 0 75 MB calciumspikesinastro 3 Channels 8 bit 65 kB 0 57 MB ciliate 3 Channels 8 bit 0 16 MB 0 56 MB A 3 Channels 8 bit 0 15 MB 0 57 MB p da autofluorescence 3 Channels 8 bit 0 41 MB 0 87 MB candidaalbicans 3 Channels 8 bit 0 17 MB cy sticmousekidney 3 Channels 8 bit 76 kB 0 75 MB AlzheimerFRETFLIM 3 Channels 8 bit 0 16 MB 0 87 MB 3 Channels 8 bit 0 39 MB 0 75 MB ChlinNIH3T 3 Channels 8 bit 0 13 MB 0 58 MB cysticproximaltubules 3 Channels 8 bit 0 11 MB 0 75 MB AlzheimerFRETFLIMs 3 Channels 8 bit 24 kB 0 43 MB bilecanaliculi 3 Channels 8 bit 0 55 MB 1 1 MB centrin 3 Channels 8 bit 88 kB Daphn 3 Channels 8 bit 89 kB 0 87 MB alzheimers 3 Channels 8 bit 0 30 MB 1 1 MB bonemarrow 3 Channels 8 bit 69 kB 0 75 MB cerebralarmyloidangio 3 Channels 8 bit 0 17 MB 0 87 MB DAPI 3 Channels 8
14. ll filters and beam splitters the channels incl settings for gain and offset and the pinhole position and diameter Frame Fast The scanning procedure can be made faster Only the laser line intensity and the Amplifier Offset are switched but no other hardware components The tracks are all matched to the current track with regard to emission filter dichroic beam splitter setting of Detector Gain pinhole position and diameter When the Line button is selected the same rules apply as for Frame Fast An additional track is added to the configuration list in the Imaging Setup Tool The maximum of four tracks can be used One track each with basic Add Track button configuration is added i e Ch 1 channel is activated all laser lines are switched off emission filters and dichroic beam splitters are set in accordance with the last configuration used m Remove button The track marked in the List of Tracks panel is deleted A A click on this arrow button will move the selected track highlighted in light grey one position upwards in the list box 42 A click on this arrow button will move the selected track highlighted in light grey one position downwards in the list box 14 08 2007 Scanning an image Setting the parameters for scanning e Select the Acquisition Mode tool from the Left Tool Area Fig 18 e Select the Frame Size as predefined number of pixels or enter your own values e g 300 x 600 in the Acquisiti
15. ment with the New button in the Action Button area or the New function in the File Menu Acquired data is not automatically saved to disc Make sure you save your data appropriately and back it up regularly The ZEN software will ask you if you want to save your unsaved images when you try to close the application Note There is no image database any more like in the earlier Zeiss LSM software versions ee a Tu Cragg Omiran Ebay See yd say ot a a ioo h Fa Flew Conic Setup Moraga ADI nil 458 477 40A 514 re bina 4i A aw r New Image mE E H fi ii mo ON P 7 7 i J cha ba CE te Fig 8 New image document in the Open Images Ares 08 2007 7 Advanced data browsing is available through the File Browser Ctrl F or trom the File Menu The File Browser can be used like the WINDOWS program file browser Images can be opened by a double click and image acquisition parameters are displayed with the thumbnails Fig 9 For more information on data browsing please refer to the detailed operating manual Fig 9 My Documents My Pictures Desktop Axiovision 4 6 Icons demo data 4D cells fromChris Nice Small LSM Demo Image Gif Movies Riken Meta images Kogure Best mdb vonZIKUP ZIKUP_ImageData mdb JenaKultur JofBioTech_CallforPapers LSM 4 2 Icons Movies Music narration zen temp install ZEN2007 AIM AIM45 Daten_Olaf Documents and Settin
16. ns of the imaging laboratory guaranteeing optimal display contrast and minimal stray light for high sensitivity detection experiments The ZEN software is optimized for a 30 TFT monitor but can also be used with dual 20 TFT setups amp Time Series Pro Mode Interval Cycle Delay Time Cycle Delay Time experiment 1 Trigge Trigger2 v x Basic Mode Time Series a Marker Cycles i Marker Interval Fig 6 Basic and Pro Mode A focus in the development of ZEN was to fulfill the needs of both basic users and microscopy specialists Both types of users will appreciate the set of intuitive tools designed to make the use of a confocal microscope from Carl Zeiss easy and fast 08 2007 5 The Pro Basic concept ensures that tool panels are never more complex than needed In Basic Mode the most commonly used tools are displayed For each tool the user can activate Pro Mode to display and use additional functionality Fig 6 Workspace Zoom E Light Path Acquisition Fig 7 ZEN Window Layout configuration More features of ZEN include e The user can add more columns to the Left Tool Area or detach individual tools to position them anywhere on the monitor To add a column drag a tool group by the title bar e g Online Acquisition to the right and a new tool column automatically opens To detach a tool click on the little icon on the very right end of the blue tool header bar Fig 7 e
17. on Mode tool Click on the Optimal button for calculation of appropriate number of pixels depending on objective N A and The number of pixels influences the image resolution w Acquisition Mode Objective Plan Apochromat 63x 1 4 Oi DIC Scan Mode Frame T Frame Size X 512 H F Find Optimal Setup Manager Laser x Imaging Setup a S E Light Path 7 Online Acquisition Pixel Dwell 1 60 psec Scan Time 3 93 sec Averaging Multidimensional Acquisition Number Bit Depth B Bit D ZStack Bleaching 7 O Time Series 3 D Scan Area Q Tile Scan 7 i Information On Experiment Fig 18 Acquisition Mode tool Note When using an Axioskop 2 FS MOT make sure you set the correct objective manually in the Acquisition Mode tool This ensures correct calibration calculation of pinhole and Z stack optimization etc The Axioskop 2 FS MOT does not automatically detect the objective lens Adjusting scan speed e Use the Scan Speed slider in the Acquisition Mode tool Fig 18 to adjust the scan speed A higher speed with averaging results in the best signal to noise ratio Scan speed 8 usually produces good results Use speed 6 or 7 for superior images Choosing the dynamic range e Select the dynamic range 8 or 12 Bit per pixel in the Bit Depth pull down in the Acquisition Mode tool Fig 18 8 Bit will give 256 gray levels 12 Bit will give 4096 gray levels Publication quality images should be ac
18. oring and exporting image data e To save your acquired or processed images click on the Save or Save As button in File Menu Fig 25 1 click on the EJ button at the bottom of the File Handling Area Fig 25 2 or click the B button in the Main Toolbar Fig 25 3 e The WINDOWS Save As window appears e Enter a file name and choose the appropriate image format Note the LSM 5 format is the native Carl Zeiss LSM image data format and contains all available extra intormation and hardware settings of your experiment e Click on the SAVE button If you close an image which has not been saved a pop up window will ask you if you want to save it Choosing yes will lead you to the WINDOWS Save As window To export image display data a single optical section in raw data format or the contents of the image display window including analysis and overlays choose Export from the File Menu In the Export window you can select trom a number of options and proceed to the WINDOWS Save As window to save the exported data to disk 08 2007 Switching off the system e Click on the File button in the Main Menu bar and then click on the Exit button to leave the ZEN software e f any lasers are still running you should shut them off now in the pop up window indicating the lasers still in use e Shut down the computer e Wait until the fan of the Argon laser has switched off e On the REMOTE CONTROL turn off the Components switch
19. quired using 12 Bit data depth 12 Bit is also recommended when doing quantitative measurements or when imaging low fluorescence intensities 08 2007 15 Setting scan averaging Averaging improves the image by increasing the signal to noise ratio Averaging scans can be carried out line by line or frame by frame Frame averaging helps to reduce photo bleaching but does not give quite as smooth of an image e For averaging select the Line or Frame mode in the Acquisition Mode tool e Select the number of lines or frames to average Adjusting pinhole size e Select the Channels tool in the Left Tool Area e Set the Pinhole size to 1 Airy unit for best compromise between depth discrimination and detection efficiency Pinhole adjustment changes the Optical Slice thickness When collecting multi channel images adjust the pinholes so that each channel has the same Optical Slice thickness This is important for colocalization studies w Channels Channel Ch Pinhole 1 00 Airy Units 116 pm section Gain Continuous Amp Offset Setup Manager 9 Laser 6 Imaging Setup E Light Path Ss LM Track Tracki L Online Acquisition gt Acquisition Mode Channels D Fous lt Stage Regions vw v 488 514 561 633 800 LS S is iS LS Transmission lt Multidimensional Acquisition D ZStack amp Bleaching Time Series Q Tile Scan i Information On Experim
20. rder to obtain some first images from his samples This Quick Guide does NOT replace the detailed information available in the full user manual or in the manual of the respective microscopes Axio Imager Axio Observer Axioskop 2 FS MOT Also this Quick Guide is written for a user who is familiar with the basics of Laser Scanning Microscopy For your safety Observe the following instructions The LSM 510 LSM 510 META LSM 510 NLO laser scanning microscope including its original accessories and compatible accessories from other manufacturers may only be used for the purposes and microscopy techniques described in this manual intended use In the Operating Manual read the chapter Safety Instructions carefully before starting operation Follow the safety instructions described in the operating manual of the microscope and X Cite 120 lamp HBO 100 mercury lamp 08 2007 1 Starting the System Switching on the LSM system e When set to ON the REMOTE CONTROL switch labeled System PC provides power to the computer This allows use of the computer and ZEN software offline Fig 1 e To completely switch on the system now press the Components switch to ON This starts the other components and the complete system is ready to be initialized by the ZEN software Switching on the X Cite 120 or the HBO 100 mercury lamp Fig 1 REMOTE CONTROL switch e Switch on the main switch of the X Cite 120 HBO 100 lamp
21. scanning of single double and triple labeling Advantage faster image acquisition Disadvantage Eventual cross talk between channels Sequential scanning of double and triple labeling line by line or frame by frame Advantage Only one detector and one laser are switched on at any one time This reduces cross talk Disadvantage slower image acquisition e Open the Imaging Setup and the Light Path tool in the Setup Manager Tool group to access the hardware control window to set up the beam path The open Light Path is shown in Fig 14 O E Light Path Acquisition LSM Track1 ld Oz BP 500 550 IR Mirror IE NFT 565 BP 650 710 IR HFT o 488 561 6 o Laser Hu Ld None 0 Imaging Setup a 4 E Light Path Pal i uisiti iti a lt Stage nnels 4 2 lt e lal Regions 7 Multidimensional Acquisition ZStack 7 l j a fal Bleaching Time Series Tile Scan 4 i Information On Experiment FT 510 LP 515 Fig 14 Light Path tool for a single track LSM 12 08 2007 e Select Channel Mode if necessary Fig 15 e Click on the LSM tab Fig 14 The Light Path tool displays the selected track configuration which is used for the scan procedure e You can change the settings of this panel using the following function elements assigning a color to the channel Select the appropriate filters and activate the channels
22. the beam path of the microscope for the port where the camera is attached for camera based image acquisition e Click on the LSM button to set the beam path for the LSM 510 system Setting up the microscope and storing settings Click on the Ocular button for direct observation Then choose the Light Path tool from the Left Tool Area Since the Ocular button has been chosen before the Oculars tab is pre selected and presented as the front tab in this tool Fig 11 E Light Path ke Selecting an objective Ocular e Open the graphical pop up menu by clicking on the Objective symbol and select the objective lens for your experiment Fig 11 Configuration e The chosen objective lens will automatically move into the beam path ofso Closed 100 63 64 EC Plan Neofluar Fi 10x 0 30 M27 Focusing the microscope for transmitted light e Open the graphical pop up menu by clicking on the Transmitted Light icon Fig 12 Focus a le e Click on the On button Set the intensity of the Halogen saa Cau lamp using the slider e Clicking outside the pop up control closes it J J V ka e Place specimen on microscope stage The cover slip must On 50 Open be facing the objective lens Remember the immersion medium if the objective chosen requires it e Use the focusing drive of the microscope to focus the Fig 11 Microscope Control window object plane e g Axio Imager Z1 e Select specimen detail by movin
23. tware package for new image acquisition and analysis The Image Processing mode ignores all hardware and activates only data handling and image processing functionality for already acquired images The Offline Demo mode reads the current hardware database but does not activate the system hardware for use Instead it simulates the system hardware for training purposes Upon clicking the Start System button the Image Processing button changes to a Cancel button Click Cancel to interrupt stop the Startup of the system After Startup the ZEN Main Application Window Fig 4 and Fig 5 opens To benefit trom all of Zen s features run the window in its Tull screen mode 08 2007 3 Application Bar Menu Bar Main Toolbar fic Ute 2 Y a i i i 6 i AN re Arpt oe O topione S Sintered KPED opam a ocmw Left Tool Area Center Screen Area hosts up to 3 Image Containers Right Tool Area Status Area Fig 4 ZEN Main Application Window after Startup with empty image container AlzheimerFRETFLIM ipo 0 87 MB AlzheemerFRETRI IMeyresinn eg C 0 43 MB Abheamners41 px 0 50 MB Torrens a Set Rrit Focus Position 0 00 ym O 2 Stack Neapsticn 0 75 MB bliecanatoul jeg 1 1 MS Multi TrackZStackFL Refi tem ec 25 MB 1 Tabs to switch between tool types 6 View Tabs 2 Action Buttons 7 Image Tabs 3 Tool Group 8 Displayed Image 4 Tool 9 File Handling Area 5 Status Bar 10 Vi
Download Pdf Manuals
Related Search