page 3 Chapter 2:Getting Started-Basic maging Functions Zoom ROI Tool Zoom ROI is a tool designed for use with fast or fixed focal length lenses.These lenses typically have lower F-stop numbers than zoom lenses,but do not have zoom capabilities.Fast lenses are most commonly used to decre acquired image's region of interest is possible using the AlphaView software interface. When the sample's region of interest is smaller than the acquisition window's and the system is not equipped with a zoom lens,the zoom ROl tool provides an ideal workaround.The Zoom ROI tool allows for software-based zooming without having to switch lenses.To use the Zoom ROI tool,focus on the image,then and Acqre ge butons to confre and aure the fh e region of interest occupies the preview windows entire nage are tool eliminates the need for cropping an image down to a desired size post-acquisition. 00 Figure 2-16:Zoom ROI Tool FluorChem HD2 and FC2User Guide
page 32 Chapter 2: Getting Started—Basic Imaging Functions FluorChem HD2 and FC2 User Guide Zoom ROI Tool Zoom ROI is a tool designed for use with fast or fixed focal length lenses. These lenses typically have lower F-stop numbers than zoom lenses, but do not have zoom capabilities. Fast lenses are most commonly used to decrease exposure times in chemiluminescence applications. However, they can also be used for other applications, such as fluorescent gels. Since fixed focal length lenses do not have zoom rings, adjusting an acquired image’s region of interest is possible using the AlphaView software interface. When the sample’s region of interest is smaller than the acquisition window’s and the system is not equipped with a zoom lens, the zoom ROI tool provides an ideal workaround. The Zoom ROI tool allows for software-based zooming without having to switch lenses. To use the Zoom ROI tool, focus on the image, then position the Zoom ROI slider until the region of interest occupies the preview window’s entire image area. Use the Expose Preview and Acquire Image buttons to configure and acquire the final image. The Zoom ROI tool eliminates the need for cropping an image down to a desired size post-acquisition. Figure 2-16: Zoom ROI Tool
Basic Imaging Functions Saving Images Once a satisfactory image has been captured,it should be saved as an original file for safe archiving.Click File>Save As or click the Save or Save As icons. Super-OD Acquisition Feature this the range of different light()levels from the sample that can be viewed as a single image beyond that of a standard image.Super-OD acquisition can produce an image that represents more than 100-fold extension of the brightness range. Camera Setup and Preriew 10 5 Figure 2-17:Super-OD Acquisitio Feature Window The AlphaView acquisition window contains a Super-OD slider bar located above the image window display. FluorChem HD2 and FC2 User Guide
Basic Imaging Functions page 33 FluorChem HD2 and FC2 User Guide Saving Images Once a satisfactory image has been captured, it should be saved as an original file for safe archiving. Click File > Save As or click the Save or Save As icons. Super-OD Acquisition Feature Super-OD™ is a process that delivers full dynamic range performance for digital imaging applications. Super-OD acquisition accomplishes this by extending the range of different light (or brightness) levels from the sample that can be viewed as a single image beyond that of a standard image. Super-OD acquisition can produce an image that represents more than 100-fold extension of the brightness range. Figure 2-17: Super-OD Acquisition Feature Window The AlphaView acquisition window contains a Super-OD slider bar located above the image window display
page 34 Chapter 2:Getting Started-Basic maging Functions Acquiring in Super-OD Acquisition Mode is in any position other than"Off".As the slider bar is moved to the right the'Super-OD Factor"will increase and be displayed. Super OD Figure-1:Super OD Slider In addition to the Super-OD Factor,the software also shows a time estimate for the total acquisition time. This time estimate is displayed as'Time x value"where Time'is the initial exposure time and the value'is the multiplier to achieve the chosen Super-OD Factor.In the example above.the Factor is set to 8.The total total acquisition time for this acquisition will be 10 milliseconds x 544=5440 milliseconds or 5.44 seconds. Initial Exposure Time for Super-OD Acquisition Whenever Super-OD is activated the acquisition interface disables the exposure time input fields and switches to Auto Exposure mode.The initial exposure time used for Super-OD is determined by a proprietary Auto Exposure algorithm.Clicking the Expose Preview button in Super-OD mode will calculate and display the initial exposure time.Clicking the Acquire button in Super-OD mode will start the Super-OD acquisition process which first determines the initial exposure value and then performs the Super-OD acquisition process automatically. Acquiring Super-OD Images Slide the Super-OD slider bar to the desired Super-OD factor Click the Acquire button.When the process is complete the resultant Super-OD image will be displayed in the AlphaView software main window. Movie Mode If kinetic,multiplex,color or chemiluminescence experime ents require the system to automatically capture several images at preset exposure times,preset time delay between images,preset lighting sources,or preset filter choices,then the AlphaView Movie Mode should be used.Movie Mode is accessible in the Camera Setup and Preview window.To launch the setup screen.click the Movie Mode button in the Camera Setup and Preview window.Movie Mode can also be accessed from Movie Mode in the ToolBox Enhancement Tools tab. FluorChem HD2 and FC2User Guide
page 34 Chapter 2: Getting Started—Basic Imaging Functions FluorChem HD2 and FC2 User Guide Acquiring in Super-OD Acquisition Mode The user may acquire a Super-OD image with a factor from 2 to 500. Choose the Super-OD factor to use by sliding the Super-OD slider bar to the right. Super-OD acquisition is activated whenever the Super-OD slider is in any position other than “Off”. As the slider bar is moved to the right the “Super-OD Factor” will increase and be displayed. Figure 2-18: Super OD Slider In addition to the Super-OD Factor, the software also shows a time estimate for the total acquisition time. This time estimate is displayed as “Time x ‘value’” where ‘Time’ is the initial exposure time and the ‘value’ is the multiplier to achieve the chosen Super-OD Factor. In the example above, the Factor is set to 8. The total acquisition time will be 544 times the initial exposure time. If the initial exposure time is 10 milliseconds, the total acquisition time for this acquisition will be 10 milliseconds x 544 = 5440 milliseconds or 5.44 seconds. Initial Exposure Time for Super-OD Acquisition Whenever Super-OD is activated the acquisition interface disables the exposure time input fields and switches to Auto Exposure mode. The initial exposure time used for Super-OD is determined by a proprietary Auto Exposure algorithm. Clicking the Expose Preview button in Super-OD mode will calculate and display the initial exposure time. Clicking the Acquire button in Super-OD mode will start the Super-OD acquisition process which first determines the initial exposure value and then performs the Super-OD acquisition process automatically. Acquiring Super-OD Images Slide the Super-OD slider bar to the desired Super-OD factor. Click the Acquire button. When the process is complete the resultant Super-OD image will be displayed in the AlphaView software main window. Movie Mode If kinetic, multiplex, color, or chemiluminescence experiments require the system to automatically capture several images at preset exposure times, preset time delay between images, preset lighting sources, or preset filter choices, then the AlphaView Movie Mode should be used. Movie Mode is accessible in the Camera Setup and Preview window. To launch the setup screen, click the Movie Mode button in the Camera Setup and Preview window. Movie Mode can also be accessed from Movie Mode in the ToolBox Enhancement Tools tab
Basic Imaging Functions page 35 Camera Setup and Preview ”:高0哥 207:a0✉四￥日 回+回 ▣中 Figure 2-19:Movie Mode Setup The Total Frames field sets the total number of individual frames in the movie.Each movie can capture maximum of 50 frames (images)and a minimum of one frame The Frame field selects the active frame currently being used for condition customization.For example,if the movie uses three images,selecting Frame I allows for lighting and filter setup of the movie's first frame: selecting Frame 2 allows for setup of the movie's second frame.All frames can be adjusted manually using this technique,or specific lighting conditions can be copied to the next frame using the Copy To Next button. Copy To Next copies all settings from the previous frame to the current frame.The Copy to End button nscopies current frame setting to the entire frame set For chemiluminescence imaging.all lighting is typically shut off and the fiter wheel is positioned for the nsche miluminescence position across all frames.In these situations,the exposure time is the only variable that changes from frame to frame.The Copy To Next button is a useful tool that optimizes setup time. The Delay field configures a predetermined delay between captured imaged for kinetic experiments.The default Delay is set for the shortest possible delay(19 ms).but can be configured up to 50 minutes between images. FluorChem HD2 and FC2 User Guide
Basic Imaging Functions page 35 FluorChem HD2 and FC2 User Guide Figure 2-19: Movie Mode Setup The Total Frames field sets the total number of individual frames in the movie. Each movie can capture maximum of 50 frames (images) and a minimum of one frame. The Frame field selects the active frame currently being used for condition customization. For example, if the movie uses three images, selecting Frame 1 allows for lighting and filter setup of the movie’s first frame; selecting Frame 2 allows for setup of the movie’s second frame. All frames can be adjusted manually using this technique, or specific lighting conditions can be copied to the next frame using the Copy To Next button. Copy To Next copies all settings from the previous frame to the current frame. The Copy to End button nscopies current frame setting to the entire frame set. For chemiluminescence imaging, all lighting is typically shut off and the filter wheel is positioned for the nschemiluminescence position across all frames. In these situations, the exposure time is the only variable that changes from frame to frame. The Copy To Next button is a useful tool that optimizes setup time. The Delay field configures a predetermined delay between captured imaged for kinetic experiments. The default Delay is set for the shortest possible delay (19 ms), but can be configured up to 50 minutes between images
page 36 Chapter 2:Getting Started-Basic maging Functions Once the movie setup is properly configured,click the Go button to begin the movie process.The system begins processing image acquisitions for each frame.When completed,the movie setup box disappears and the Toolbox window automatically configures to the Movie tools.This allows for movie playback,saving,load- ing,or new movie recording. The movie display buttons offer the following functionality REC Launches the Camera Setup and Preview window configured to Movie Mode. PLAY Displays the movie in a continuous loop. STOP Stops the movie at the current frame display. PAUSE Pauses movie playback at a user defined image. REW(rewind) Rewinds the movie to the first image. REV(reverse) Plays the movie in a continuous loop in reverse. FWD(forward) Forwards the movie to the last image. DEL(delete) Deletes the movie on the display. NOTE:This function does not delete movies saved to the hard drive. LOAD Loads a previously saved movie NOTE:This function does not load individual images previously captured in nor- mal capture mode SAVE Saves a movie of images. Saving An Individual Image from a Movie After you load,play,and stop a movie at the desired image,it is possible to save the individual image seenon format on the local or network drives FluorChem HD2 and FC2 User Guid
page 36 Chapter 2: Getting Started—Basic Imaging Functions FluorChem HD2 and FC2 User Guide Once the movie setup is properly configured, click the Go button to begin the movie process. The system begins processing image acquisitions for each frame. When completed, the movie setup box disappears and the Toolbox window automatically configures to the Movie tools. This allows for movie playback, saving, loading, or new movie recording. The movie display buttons offer the following functionality: Saving An Individual Image from a Movie After you load, play, and stop a movie at the desired image, it is possible to save the individual image seen on the screen. Use the SAVE AS button located on the tool bar to save the image in the desired location and file format on the local or network drives. REC Launches the Camera Setup and Preview window configured to Movie Mode. PLAY Displays the movie in a continuous loop. STOP Stops the movie at the current frame display. PAUSE Pauses movie playback at a user defined image. REW (rewind) Rewinds the movie to the first image. REV (reverse) Plays the movie in a continuous loop in reverse. FWD (forward) Forwards the movie to the last image. DEL (delete) Deletes the movie on the display. NOTE: This function does not delete movies saved to the hard drive. LOAD Loads a previously saved movie. NOTE: This function does not load individual images previously captured in normal capture mode. SAVE Saves a movie of images