NIS-Elements F User s Guide. Laboratory Imaging

Transcription

1 NIS-Elements F 2.20 User s Guide Laboratory Imaging

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3 No part of this manual may be reproduced or transmitted except with the written permission of Laboratory Imaging, Ltd. Information within this manual is subject to change without notice. Changes, technical inaccuracies and typographical errors will be corrected in subsequent editions of this manual. Revision: 1.0, April 4, 2006

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5 Table of contents Welcome to NIS-Elements F 5 Quick Start 7 1 Installation 7 2 Connecting Camera 7 3 Starting Application 7 4 Troubleshooting 7 Main Window 9 Operation modes 9 Areas of interest 10 Documents treatment 10 Zooming 10 Tools 11 Camera settings 11 Working with camera 12 File Menu 13 Edit Menu 15 Camera Menu 17 Preprocess Menu 19 View Menu 21 Help Menu 23 Camera Settings for DXM1200(F) 25 Camera Settings for DXM1200C 27 Camera Settings for DS-1QM 31 Settings for DS-U1 cameras 35 Settings for DS-U2 cameras 39 Camera Settings for DQC-FS 45 Camera Settings for Cascade 512B 49 Camera Settings for Coolsnap ES 53

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7 Welcome to NIS-Elements F This software is supplied along with laboratory digital cameras. It enables user to run the camera, capture and save images, make basic image processing etc. It is also a preview of what is provided with the higher versions of NIS-Elements software: NIS-Elements Basic Research and NIS-Elements Advanced Research. Page 5

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9 Quick Start We recommend not to plug the camera unit into the computer before the installation. 1 Installation - Insert the installation CD into your CD-ROM drive - Locate the setup.exe file and run it. - The install shield that appears will guide you through the installation process. - When asked, select the correct camera type. - Finish the setup. 2 Connecting Camera - Connect the camera head to the control unit. - Connect the control unit to the computer. - Switch the control unit ON. Wait until a solid green light shines on the control unit. 3 Starting Application - Click the desktop icon or go to the Start menu > Programs > NIS-Elements F > Run NIS-Elements F - The main window appears, ready to be used. - Click the Live button, and you get the live camera signal. 4 Troubleshooting It is not possible to run the application without a camera connected to the system. If the application is unable to start correctly, the reason may be: - The camera is not plugged into the computer properly or is switched OFF. - Check if the camera cable is attached to the computer and if the camera (or the control unit) is switched ON. - The camera system drivers are not installed or are out of date. - The camera drivers should have been installed during the setup. However, some unexpected error might occur or you might have selected a wrong camera type by mistake. Try to: - Re-install the application. - Install the camera drivers directly from the installation CD. - Restart the computer. - If none of these steps helps, please, locate the lxapp.log file in the installation directory Page 7

10 Page 8 and send it by mail to your local Nikon dealer. The technical support team will give you a hand.

11 Main Window Operation modes This mode enables you to click on the image and move it within the image window. Zooming can be performed by a mouse wheel. This mode changes the cursor to an interactive zoom tool that magnifies a circular part of the current image when clicked. Page 9

12 This mode enables you to insert annotation objects into pictures according to the Tools settings. Areas of interest This tool defines the area of the live image that is used to determine auto exposure parameters applied to the camera. This tool defines the area of the live image that is used to determine the auto focus parameters. Place the rectangle to where you would like to focus. Documents treatment Opens an image file stored on harddisk. This command can save a captured or modified picture on hard disk. Opens a printing dialog window that enables you to print the current image if a printer is present. This button creates a basic image report. It makes a pdf file that contains the current image, its name, path, dimensions, and the file date. Zooming Adjusts the zoom factor to view exactly the whole image as big as possible. Adjusts the zoom factor to view the current image on the 1:1 scale. Increases the zoom factor of the current image. Page 10

13 Decreases the zoom factor of the current image. Enables to zoom the image within a range of 10%-1200%. Tools Displays a scale in the bottom right corner of the image. There are seven scales for different objective magnifications (2, 4, 10, 20, 40, 60, 100 times) predefined. A number of annotation objects can be inserted into the image. This button indicates whether the line, arrow, text, rectangle, or circle mode is active. Pressing the button toggles between the Annotate and Point operation modes. An intensity profile of a user defined cross section can be displayed in the image. Camera settings Auto Exposure modes The number of available AE modes differs according to the camera type. - AE F (Focus) - Calculates optimal combination of Exposure time and Gain in order to achieve optimal exposure. High frame rate - which is priority in this mode - limits the Exposure time. - AE P (Program) - Calculates optimal combination of Exposure time and Gain in order to achieve optimal exposure. Image quality is the priority, therefore longer exposure times are chosen. - AE S (Shutter) - Calculates optimal Gain for a given Exposure time. Exposure Compensation If the current autoexposure mode gives an image that is too bright or too dark, use the exposure compensation. Click the High button to get a brighter image and the Low button to get a darker image. Auto Exposure Area Click the Show button to display and activate the auto exposure area. The area is used to determine auto exposure parameters applied to the camera. Clicking on the arrow on the left side will cause the AE area size to be selected and the position reset to the center. Page 11

14 Advanced The advanced settings such as noise reduction and others can be set. The settings available depend on the camera type. Working with camera Displays the live image in the picture window. Freezes and grabs the live camera image into the picture window as the Frozen document. This command captures Live or Frozen image to the application window. It creates a new document called Captured. This command captures Live or Frozen image and saves it automatically and names it according to the current settings (File menu > Options). Page 12

15 File Menu The menu contsists of the following commands: Open This command loads a specified image file according to selected options. Save This command saves modifications of the current image onto a disk. Save As This command saves an image onto a disk (in case of a new image) or enables you to save the image under a different name (in case the image has already been saved). Reload This command reloads the current image that has been modified, and returns the it to the last saved state. A confirmation dialog appears. Print This command opens a printing dialog window that enables you to print the current image (if a printer is present). Send This command opens a default application and attaches the current image automati- Page 13

16 cally. Report This command creates a basic image report. It makes a new pdf file that contains the current image, its name, path, dimensions, and the file date. Options Opens the options dialog box. Working folder - is the folder used as default by the Open, Save, Save As commands. Although, pictures of the working folder are displayed inside the bottom preview window. Auto Capture - defines a prefix and the format of the files captured and saved via the Auto Capture command. Calibration - here you can select the units used for calibration of the objective and the units precision. Exit This command quits the application. Page 14

17 Edit Menu The menu contsists of the following commands: Undo This command displays the current image as it appeared before the most recent transformation. Redo This command returns the image back to the state that was displayed before the Undo command was called. Resize Using this command it is possible to adjust image dimensions. You can enter the new size as an exact value in pixels or in percentage of the current image size. Crop This command cuts off everything outside a selected image area. Selecting the command invokes the Crop cursor. Frame the area you want to crop by the red rectangle. Use the white squares on the rectangle edges to adjust its size, and drag it with the left mouse button to set its position. Finish the command with the right click or by pressing Enter. Page 15

18 Copy Copies the whole image into clipboard. Flip Horizontally This command allows you to mirror all parts of the current image horizontally. Flip Vertically This command allows you to mirror all parts of the current image vertically. Rotate Right This command performs a rotation by -90. Rotate Left This command performs a rotation by 90. File Merge This command can combine color planes, stored in separate files, into one RGB image. Page 16

19 Camera Menu The menu contsists of the following commands: Live, Freeze, Capture, Auto Capture These commands correspond to the main window Camera section buttons. Please see the Main window description - Working with camera. Calibrate This command can modify the calibration of the live camera signal. To calibrate the current objective: - Click the Calibrate command. - A dialog window appears. Select the calibration lines orientation. - Place the parallel lines to the image window. - Another dialog window appears. - Fill in the real distance between the lines and confirm the calibration by OK. - The objective is calibrated. Page 17

20 Page 18 Using this command, the current objective (the one selected in the Tools section of the main application window) calibration is modified and saved. The other six objectives calibrations change accordingly.

21 The menu contsists of the following commands: Preprocess Menu Contrast This command enhances contrast of the current color image. The Auto contrast and the Gamma correction can be applied. White Balance This command balances and adjusts red, green and blue components of the current color image. The White Balance Adjustment dialog window appears. One or two circular probes appear in the picture window at the same time. One probe is used for definition of the white level, the other for the black level. Change Saturation This command changes the saturation of a color image. It can be increased or decreased within the 200% range (+100, -100) Change Hue This command transforms a color image by a hue shift. Page 19

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23 View Menu The menu contsists of the following commands: Zoom (submenu) This submenu contains commands corresponding to the zoom section buttons. Please see the Main window description - Zooming. Full Screen This command switches the application to the full screen view. All toolbars and menus are hidden and only the current image is displayed. To quit the full screen view, pres Esc. Scale (submenu) The Show Scale Bar command displays a rough scale in the bottom right corner of the image window. There are seven predefined scales for different objective magnifications (2, 4, 10, 20, 40, 60, 100 times). These can be selected from the Tools section of the main application window Page 21

24 The Scale Bar Properties such as scale color, background color, thickness, rotation, or units can be defined by the second command of this submenu. Operation Mode (submenu) This submenu contains commands corresponding to the Operation modes section buttons. Please see the Main window description - Operation modes. Page 22

25 Help Menu The menu contsists of the following commands: Contents This command displays the content of the electronic documentation. Search This command opens the electronic help file at the search page. About This command displays the version of the software, and the copyright information. Page 23

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27 Camera Settings for DXM1200(F) Live Format A camera format used with the Live command. It is optimized to give as many frames per second as possible in low resolution. This mode increases the gain and uses short exposure times. Format For Capture A camera format used with the Capture command. It gives pictures in high resolution, but the frame-rate is low. This mode sets the gain as low as possible and extends the exposure time. The 1280x1024,Fine(SW) mode The DXM1200F camera does not provide the image resolution of 1280x1024 px. The (SW) sign means that the images are captured in maximal camera resolution and then resampled by the software to the resolution indicated. This feature might be useful e.g. while saving the capacity of a hard drive. Preview in Capture Quality Starts live preview with format specified in the Format For Capture combo box. Page 25

28 Auto White Automatically adjusts gains of the red, green and blue components to get the color neutral white either in the whole picture window or in the probe. You can switch the probe on/off in the application toolbar. Auto Exposure Initiates the auto exposure mode. The software analyzes incoming images and adjusts the exposure to get the best results. Exposure Controls the exposure time of the camera. Influences the frame rate. On DXM1200/1200F you get maximally 12 Fps on 640x512 resolution. Gain Influences the image contrast. It is a multiplication constant that controls the color dynamics (especially with lower illumination levels). A possible contrast enhancement procedure is to set a lower offset and then to raise the gain. Red, Green, Blue - Gains for Red, Green, and Blue components. Offset Sets the brightness of the image. It is a constant additive (positive or negative) changing all pixel values. With negative offset value the dark image areas become black. In the case of fluorescence microscopy the appropriate offset setting creates a black background and thus helps to enhance (together with gain or illumination enhancement) the contrast. This property is software-implemented. Gamma Coefficient of a gamma correction, the grabber automatically performs gamma correction on a live image. Gamma correction maps the intensity of a video signal exponentially with a parameter gamma. For gamma < 1, you get the more sensitive information for the image parts of low intensities whereas for gamma > 1 image parts of higher intensities are enhanced. This property is software-implemented. Using the AE Area The AE Area enables you to determine an image area that serves as the data source for the AWB (auto white balance) and the AE (Auto Exposure) features of the cameras. When the AE Area functionality is not supported, the AWB and AE algorithms are computed from the whole image. When the AE Area is ON: - The Auto Exposure computed from within the AE Area is supported. - The Auto White Balance computed from within the AE Area is supported. Page 26

29 Camera Settings for DXM1200C Sets the video capture board parameters up. Live Format A camera format used with the Live command. It is optimized to give as many frames per second as possible in low resolution. This mode increases the gain and uses short exposure times. Format For Capture A camera format used with the Capture command. It gives pictures in high resolution, but the frame-rate is low. This mode sets the gain as low as possible and extends the exposure time. Provided image resolutions Live Format 686x512, Focus - very fast mode for focusing1372x1024, Focus - this is the actual resolution of the DXM1200C CCD chip. The Format for Capture 1372x1024, Focus - the resulting image is a single frame of the camera.1372x1024, Fine(SW) - the image is captured in a maximal resolution and resampled by the software.4116x Using this mode, three frames are acquired. The CCD is shifted by a sub-pixel distance Page 27

30 after each frame by a precise piezo shift (inbuilt inside the camera). The resulting image is combined from the three frames. Preview in Capture Quality Starts live preview with format specified in the Format For Capture combo box. Auto White Automatically adjusts gains of the red, green and blue components to get the color neutral white either in the whole picture window or in the probe. You can switch the probe on/off in the application toolbar. This feature is available in the color mode only. Auto Exposure Performes the automatic exposure. The software analyzes incoming images and adjusts the exposure to get the best results. Cont. (Continuous AE) This checkbox turns the autoexposure mode ON. If checked, the automatic exposure is being performed continuously. AE Compensation In automatic exposure mode the compensation affects how optimum exposure settings (Exposure time and Gain) are calculated. The Compensation value is expressed in Exposure Values (EV). Setting Compensation to EV makes the image twice brighter (e.g. doubles the Exposure time or Gain). Exposure Controls the exposure time of the camera. Influences the frame rate. On DXM1200C you get maximally about 15 Fps on 686x512 resolution. Gain The image can be enhanced by gain. There are three possible settings: Normal, High, Max. Offset Sets the brightness of the image. It is a constant additive (positive or negative) changing all pixel values. With negative offset value the dark image areas become black. In the case of fluorescence microscopy the appropriate offset setting creates a black background and thus helps to enhance (together with gain or illumination enhancement) the contrast. This property is software-implemented. Gamma A coefficient of a gamma correction, the grabber automatically performs gamma correction on a live image. Gamma correction maps the intensity of a video signal exponentially to the Page 28

31 gamma parameter. For gamma < 1, you get the more sensitive information for the image parts of low intensities whereas for gamma > 1 image parts of higher intensities are enhanced. The maximum gamma value of 1 is allowed on DXM1200C. Commands Black Balance Automatically adjusts gains of the red, green and blue components to get the color neutral black. CCD Cooling DXM1200C camera has an inbuilt cooler. When it is on, it helps to reduce the dark current noise and enhances the live image. The cooler can be switched OFF by unchecking the CCD Cooling option. This feature is available in the color mode only. Set Defaults This command resets all camera settings to default values. The settings can be saved and loaded under miscellaneous names. Using the AE Area The AE Area enables you to determine an image area that serves as the data source for the AWB (auto white balance) and the AE (Auto Exposure) features of the cameras. When the AE Area functionality is not supported, the AWB and AE algorithms are computed from the whole image. When the AE Area is ON: - The Auto Exposure computed from within the AE Area is supported. - The Auto White Balance computed from within the AE Area is supported. Page 29

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33 Camera Settings for DS-1QM Live Fast / Quality Capture formats Indicates which mode is actually running in the document window. The Quality Capture mode is always used for capturing. The Format combo box Sets the output image format. Formats differ in the image size, intensity, capturing method and consequently in frame rate (frames per second - fps). Smaller resolutions are achieved using Binning (integrating more pixels together) therefore intensity changes. AE Settings in Commands menu affect how auto exposure is performed (see below) x 1000 (max 15 fps) - base chip resolution,500 x 500 (max 30 fps) - image has same brightness as the baseresolution but significantly less noise,250 x 250 (max 50 fps) - image is two times brighter than in the baseresolution,125 x 125 (max 90 fps) - image is eight times brighter than in the baseresolution. Auto Exposure The Auto Exposure button runs a single auto exposure. Correct exposure is calculated for both Live Fast and Quality Capture modes taking intoaccount the Gain settings and possibly different format intensities. If theexposure calculation should exceed twenty seconds (due to long exposuretime or averaging), the process terminates and outputs a possibly wrongexposure time. Exposure Time Controls the exposure time. It is the time of charge accumulation in CCD`s basic element, between frames. The longer the exposure is, the lighter the image you ll get. Note, that long exposure times lowers the frame rate. Page 31

34 Averaging The Averaging is a commonly used technique of decreasing noise in the image. In this method 2, 4, 8 or 16 consecutive frames are averaged together. Select the number of frames from the combo box. Temperature and Fan Shows temperature of the CCD chip in relation to the temperature outside. The Fan icon button activatesthe CCD chip cooling.! (next to the Format For Live combo box) Let s explain this on an example: Assuming your camera settings match our example picture (above), the Live mode produces data of much higher intensity than the Capture mode. In order to display the live image with the same intensity as the captured image would have, the system shortens the exposure time of the Live mode. However, if the Live binning is too high, and the Capture exposure time too short, the Live Exposure time cannot be shortened any more. At that time, The! button turns red to remind you. Press it, and the Live mode binning will be automatically changed to a suitable value. Commands > Advanced Setup Live Acceleration Would you like to increase the frame rate of the live signal? Move the slider to a desired multiplier. The system then automatically shortens the exposure time (so the frame rate rises), and the loss of intensity is compensated by gain (software multiplication). Target Maximal Intensity Restricts the maximal image intensity after the auto exposure was applied. The value represents a percentage of the whole camera dynamic range. Maximal Exposure This is a safeguard of the time of the Auto Exposure. For quick exposure, it is convenient not to set this value too high. Overillumination Tolerance Sets how many pixels should be white after the Auto exposure is performed. Use lower values Page 32

35 (0.01%) for very bright (shining) objects (fluorescence). For common brightfield, even 1% may be a good value. Optionally, you can define the absolute number of white pixels. Using the AE Area The AE Area enables you to determine an image area that serves as the data source for the AWB (auto white balance) and the AE (Auto Exposure) features of the cameras. When the AE Area functionality is not supported, the AWB and AE algorithms are computed from the whole image. When the AE Area is ON: - The Auto Exposure computed from within the AE Area is supported. - The Auto White Balance computed from within the AE Area is not supported. Page 33

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37 Settings for DS-U1 cameras Fast (Focus) Format and Quality (Capture) Format Sets the output image format. Formatsdiffer in the image size, capturing method and consequently in framerate (frames per second - fps). The higher the resolution is, the lower frame rate can be achieved. Available resolutions depend on the camera head used. 5 megapixel camera heads: 640 x 476 Central (max 15 fps) - image is taken from the central part ofthe chip, 640 x 480 Normal (max 7.5 fps) - image covers the whole chip, 1280 x 960 Fine (max 3.75 fps) - image covers the whole chip and isfour times more sensitive than the 1280 x 960 normal, 1280 x 960 Normal (max 3.75 fps) - image covers the whole chip 2560 x 1920 Fine (max 1.5 fps) - image covers the whole chip 2 megapixel camera heads: 800x560 Central - image is taken from the central part of the chip 800x600 Fine - image covers the whole chip 1600x1200 Fine - image covers the whole chip Page 35

38 Exposure Mode Exposure mode sets how the Exposure time(shutter speed) is calculated. There are two options: Auto Exposure - the exposure time is calculated automatically. Manual Exposure - you can set the exposure time manually. Exposure Controls the exposure time (the shutter speed), which is expressed inversely (1/x second). It is thetime of charge accumulation in a CCD`s basic element, between frames. Increasing the Exposure timeincreases the brightness of the image and increases the quality (less gain less noise)and decreases frame rate (fps). Enabled only when Exposure mode is setto ME or AE S. Gain Controls the Gain (Sensitivity) of thecamera. Increasing the Gain increases brightness of the image, decreases quality (more gain more noise) and increases (indirectly -enables shorter Exposure time) frame rate (fps). Enabled only whenexposure mode is setto ME. AE Compensation In automatic exposure mode thecompensation affects how optimum exposure settings (Exposure timeand Gain) are calculated. The Compensation value is expressed inexposure Values (EV). Setting Compensation to EV makes theimage twice brighter (e.g. doubles the Exposure time or Gain) which maybe useful for brightfield. AE Lock Causes the automatic exposure to lockactual exposure settings (Exposure time and Gain). Contrast Affects dynamics of how the luminosity isrendered. There are several modes for different illumination (contrast)scene situation. Auto White balance (color camera only). The Auto White button calculates theright values in order to obtain the white color correctly. The Auto White givesthe best results on color neutral (gray) scenes. Page 36

39 Advanced > Advanced Setup Mode Defines how the Auto Exposure isperformed. Average is suitable for brighfield and Peak is betterused for darkfield. Area Defines the size of central area which istaken into account for Auto exposure. Noise Reduction (NR) If activated, it reduces noise generatedby higher gain. NR works on interframe basis so you may notice someshadows on moving objects. Sharpness Affects how sharply edges appear. Too muchsharpness leads to oversaturated edges. Offset Is anadditive constant (positive or negative) that changes all pixel valuesby the same amount (the Offset value).it is used to set black signal to zero. That is when no light isfalling on the chip all pixels should have value of zero. In thecase of fluorescence microscopy the appropriate offset setting createsa black background and thus helps to enhance (together with Gain orillumination enhancement) the contrast. Saturation (color camera only) The amount of saturation determines how colors are rendered. Moresaturation produces richer colors. Less saturation makes the colorsgray. Hue (color camera only) Hue shifts colors across the rainbow. White Balance (color camera only) Gains for Red, Green, and Blue image components. Using the AE Area The AE Area enables you to determine an image area that serves as the data source for the AWB (auto white balance) and the AE (Auto Exposure) features of the cameras. When the AE Area functionality is not supported, the AWB and AE algorithms are computed from the whole image. Page 37

40 Page 38 When the AE Area is ON: - The Auto Exposure computed from within the AE Area is not supported. - The Auto White Balance computed from within the AE Area is not supported. The DS-U1 cameras compute the exposure time from the central area of the CCD chip, no matter if the Probe is ON or OFF. The size of the area can be adjusted in the Advanced setup (described above) to Large, Medium, or Small. The probe then serves as the data source only for LUTs and a histogram.

41 Settings for DS-U2 cameras Mode - Normal/Binning The Binning mode provides only one image resolution in contrast with considerably enhanced CCD chip sensitivity. The Normal mode offers all resolutions of the current camera head. Fast (Focus) Format and Quality (Capture) Format Sets the output image format. Formatsdiffer in the image size, capturing method and consequently in framerate (frames per second - fps). The higher the resolution is, the lower frame rate can be achieved. Available resolutions depend on the camera head used. Page 39

42 5 megapixel camera heads: 640 x 476 Central (max 15 fps) - image is taken from the central part ofthe chip, 640 x 480 Normal (max 7.5 fps) - image covers the whole chip, 1280 x 960 Fine (max 3.75 fps) - image covers the whole chip and isfour times more sensitive than the 1280 x 960 normal, 1280 x 960 Normal (max 3.75 fps) - image covers the whole chip 2560 x 1920 Fine (max 1.5 fps) - image covers the whole chip 2 megapixel camera heads: 800x560 Central - image is taken from the central part of the chip 800x600 Fine - image covers the whole chip 1600x1200 Fine - image covers the whole chip Exposure Mode Exposure mode sets how the Exposure time(shutter speed) is calculated. There are two options: Auto Exposure - the exposure time is calculated automatically. Manual Exposure - you can set the exposure time manually. Exposure Controls the exposure time (the shutter speed), which is expressed inversely (1/x second). It is thetime of charge accumulation in a CCD`s basic element, between frames. Increasing the Exposure timeincreases the brightness of the image and increases the quality (less gain less noise)and decreases frame rate (fps). Enabled only when Exposure mode is setto ME or AE S. Gain Controls the Gain (Sensitivity) of thecamera. Increasing the Gain increases brightness of the image, decreases quality (more gain more noise) and increases (indirectly -enables shorter Exposure time) frame rate (fps). Enabled only whenexposure mode is setto ME. AE Compensation In automatic exposure mode thecompensation affects how optimum exposure settings (Exposure timeand Gain) are calculated. The Compensation value is expressed inexposure Values (EV). Setting Compensation to EV makes theimage twice brighter (e.g. doubles the Exposure time or Gain) which maybe useful for brightfield. AE Lock Causes the automatic exposure to lockactual exposure settings (Exposure time and Gain). Contrast Affects dynamics of how the luminosity isrendered. There are several modes for different illumination (contrast)scene situation. Page 40

43 Auto White balance (color camera only). The Auto White button calculates theright values in order to obtain the white color correctly. The Auto White givesthe best results on color neutral (gray) scenes. Scene Mode There are eight presets of camera settings optimized for the specific usage. You can reset the settings by the Reset button. Industrial microscopy: - W - Wafer IC-Chip - M - Metal or ceramic - C - Circuit boards Bioscience microscopy: - F - Dark-field fluorescence - B - Bright field - D - DIC or phase contrast - H - Hematoxilin -Eosin - E - Enzyme - Antibody Commands > ROI Use Current ROI The currently defined region of interest, set by the Define command, can be switched ON/ OFF by checking this item. Define... To restrict observed scene only to a part of CCD chip: - Click the Define command. A red rectangle appears on the screen. - Double click inside the rectangle to toggle between two pre-defined sizes of ROI. - Drag ROI to the intended position. - Right click into the image. The Use Current ROI is automatically switched ON. The scene changes. Page 41

44 Commands > Advanced Mode Defines how the Auto Exposure is performed. Average is suitable for brighfield and Peak is better used for darkfield. Noise Reduction (NR) If activated, it reduces noise generatedby higher gain. NR works on interframe basis so you may notice someshadows on moving objects. Sharpness Affects how sharply edges appear. Too muchsharpness leads to oversaturated edges. Offset Is anadditive constant (positive or negative) that changes all pixel valuesby the same amount (the Offset value).it is used to set black signal to zero. That is when no light isfalling on the chip all pixels should have value of zero. In thecase of fluorescence microscopy the appropriate offset setting createsa black background and thus helps to enhance (together with Gain orillumination enhancement) the contrast. Saturation (color camera only) The amount of saturation determines how colors are rendered. Moresaturation produces richer colors. Less saturation makes the colorsgray. Hue (color camera only) Hue shifts colors across the rainbow. Preset The scene mode can be selected from this combo box. White Balance (color camera only) Gains for Red, Green, and Blue image components. Page 42

45 Using the AE Area The AE Area enables you to determine an image area that serves as the data source for the AWB (auto white balance) and the AE (Auto Exposure) features of the cameras. When the AE Area functionality is not supported, the AWB and AE algorithms are computed from the whole image. When the AE Area is ON: - The Auto Exposure computed from within the AE Area is supported. - The Auto White Balance computed from within the AE Area is supported. Page 43

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47 Camera Settings for DQC-FS Format For Live Camera format used in live command. Format For Capture Camera format used in capture command. Auto Exposure Initiates the autoexposure mode. Software analyzes incoming image and adjusts exposition and gain to get best results. Exposure Time Controls the exposure time of the camera. Available exposure time range depends on selected Format and Readout Speed. Multiplier Controls on-chip multiplication gain. Multiplication factor is adjusted in 4095 steps. There is complex relationship (approximated to an exponential) between set value and level of multiplication. Readout Speed Speed of digitization influences frame rate and image quality. Higher frequency increase frame rate, lower frequency offers better image quality. Page 45

48 Conversion Gain This is a hardware gain. Commands > Advanced Setup Live Acceleration Would you like to increase the frame rate of the live signal? Move the slider to a desired multiplier. The system then automatically shortens the exposure time (so the frame rate rises), and the loss of intensity is compensated by gain (software multiplication). Target Maximal Intensity Restricts the maximal image intensity after the auto exposure was applied. The value represents a percentage of the whole camera dynamic range. Maximal Exposure This is a safeguard of the time of the Auto Exposure. For quick exposure, it is convenient not to set this value too high. Overillumination Tolerance Sets how many pixels should be white after the Auto exposure is performed. Use lower values (0.01%) for very bright (shining) objects (fluorescence). For common brightfield, even 1% may be a good value. Optionally, you can define the absolute number of white pixels. Page 46

49 Commands > Advanced Camera Settings Trigger Mode Sets the exposure method. - Internal - The exposure time and the beginning of each frame acquisition is controlled by NIS-Elements settings. - Strobe - In this mode, the beginning of each frame exposure is being controlled by external signal. - Bulb - The exposure time and frames timing are controlled by external signal connected to the camera. Desired Temperature Sets the target temperature of the CCD chip. Clear Cycles This option enables you to decide, how many times the CCD chip should be cleared (reset). The set Clear Mode determines how often it is performed. Clear Mode Sets how often the CCD chip will be reset. Please, see the camera user manual for further details. These options are available: - Automatic - Never - Pre-exposure - Pre-sequence - Post-sequence - Pre and Post-sequence - Pre-Exposure and Post-Sequence Sensor Mode This option sets the method of reading the CCD chip. - Automatic - This option makes the system to choose the best mode. - Normal - Please, see the camera user manual for further details. - Alternate Normal Page 47

50 Keep Overlapped If checked. The Clear Mode and Sensor Mode boxes are set to Automatic and become disabled. This mode produces maximal frame rate and decreases the list of exposure times (in the camera settings window). Only the times of 1 frame and longer become available. Using the AE Area The AE Area enables you to determine an image area that serves as the data source for the AWB (auto white balance) and the AE (Auto Exposure) features of the cameras. When the AE Area functionality is not supported, the AWB and AE algorithms are computed from the whole image. When the AE Area is ON: - The Auto Exposure computed from within the AE Area is supported. - The Auto White Balance computed from within the AE Area is not supported. Page 48

51 Camera Settings for Cascade 512B Sets up video capture board parameters. Format For Live Camera format used in live command. Format For Capture Camera format used in capture command. Auto Exposure Initiates the autoexposure mode. Software analyzes incoming image and adjusts exposition and gain to get best results. Exposure Time Controls the exposure time of the camera. Available exposure time range depends on selected Format and Readout Speed. Multiplier Controls on-chip multiplication gain. Multiplication factor is adjusted in 4095 steps. There is complex relationship (approximated to an exponential) between set value and level of multiplication. Readout Speed Speed of digitization influences frame rate and image quality. Higher frequency increase frame rate, lower frequency offers better image quality. Page 49

52 Conversion Gain This is a hardware gain. Commands > Advanced Setup Live Acceleration Would you like to increase the frame rate of the live signal? Move the slider to a desired multiplier. The system then automatically shortens the exposure time (so the frame rate rises), and the loss of intensity is compensated by gain (software multiplication). Target Maximal Intensity Restricts the maximal image intensity after the auto exposure was applied. The value represents a percentage of the whole camera dynamic range. Maximal Exposure This is a safeguard of the time of the Auto Exposure. For quick exposure, it is convenient not to set this value too high. Overillumination Tolerance Sets how many pixels should be white after the Auto exposure is performed. Use lower values (0.01%) for very bright (shining) objects (fluorescence). For common brightfield, even 1% may be a good value. Optionally, you can define the absolute number of white pixels. Page 50

53 Commands > Advanced Camera Settings Trigger Mode Sets the exposure method. - Internal - The exposure time and the beginning of each frame acquisition is controlled by NIS-Elements settings. - Strobe - In this mode, the beginning of each frame exposure is being controlled by external signal. - Bulb - The exposure time and frames timing are controlled by external signal connected to the camera. Desired Temperature Sets the target temperature of the CCD chip. Clear Cycles This option enables you to decide, how many times the CCD chip should be cleared (reset). The set Clear Mode determines how often it is performed. Clear Mode Sets how often the CCD chip will be reset. Please, see the camera user manual for further details. These options are available: - Automatic - Never - Pre-exposure - Pre-sequence - Post-sequence - Pre and Post-sequence - Pre-Exposure and Post-Sequence Sensor Mode This option sets the method of reading the CCD chip. - Automatic - This option makes the system to choose the best mode. - Normal - Please, see the camera user manual for further details. - Alternate Normal Page 51

54 Keep Overlapped If checked. The Clear Mode and Sensor Mode boxes are set to Automatic and become disabled. This mode produces maximal frame rate and decreases the list of exposure times (in the camera settings window). Only the times of 1 frame and longer become available. Using the AE Area The AE Area enables you to determine an image area that serves as the data source for the AWB (auto white balance) and the AE (Auto Exposure) features of the cameras. When the AE Area functionality is not supported, the AWB and AE algorithms are computed from the whole image. When the AE Area is ON: - The Auto Exposure computed from within the AE Area is supported. - The Auto White Balance computed from within the AE Area is not supported. Page 52

55 Camera Settings for Coolsnap ES Sets up video capture board parameters. The Camera Settings dialog box appears. Format For Live Camera format used in live command. Format For Capture Camera format used in capture command. Auto Exposure Initiates the autoexposure mode. Software analyzes incoming image and adjusts exposition and gain to get the best results. Exposure Time Controls the exposure time of the camera. Page 53

56 Commands > Advanced Setup Live Acceleration Would you like to increase the frame rate of the live signal? Move the slider to a desired multiplier. The system then automatically shortens the exposure time (so the frame rate rises), and the loss of intensity is compensated by gain (software multiplication). Target Maximal Intensity Restricts the maximal image intensity after the auto exposure was applied. The value represents a percentage of the whole camera dynamic range. Maximal Exposure This is a safeguard of the time of the Auto Exposure. For quick exposure, it is convenient not to set this value too high. Overillumination Tolerance Sets how many pixels should be white after the Auto exposure is performed. Use lower values (0.01%) for very bright (shining) objects (fluorescence). For common brightfield, even 1% may be a good value. Optionally, you can define the absolute number of white pixels. Commands > Advanced Camera Settings Trigger Mode This combo box belongs to other Photometrics cameras. Desired Temperature This combo box belongs to other Photometrics cameras. Clear Cycles This option enables you to decide, how many times the CCD chip should be cleared (reset). The set Clear Mode determines how often it is performed. Page 54

57 Clear Mode Sets how often the CCD chip will be reset. Please, see the camera user manual for further details. These options are available: - Automatic - Never - Pre-exposure - Pre-sequence - Post-sequence - Pre and Post-sequence - Pre-Exposure and Post-Sequence Sensor Mode This option sets the method of reading the CCD chip. - Automatic - This option makes the system to choose the best mode. - Normal - Please, see the camera user manual for further details. - Alternate Normal - Please, see the camera user manual for further details. Keep Overlapped If checked. The Clear Mode and Sensor Mode boxes are set to Automatic and become disabled. This mode produces maximal frame rate and decreases the list of exposure times (in the camera settings window). Only the times of 1 frame and longer become available. Using the AE Area The AE Area enables you to determine an image area that serves as the data source for the AWB (auto white balance) and the AE (Auto Exposure) features of the cameras. When the AE Area functionality is not supported, the AWB and AE algorithms are computed from the whole image. When the AE Area is ON: - The Auto Exposure computed from within the AE Area is supported. - The Auto White Balance computed from within the AE Area is not supported. Page 55

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