Acquiring Images Using SimplePCI with the DVC-1412 or DVC-1312

• Log on to the computer
  • enter username (and press tab to move to-)
  • enter password (case is important)
  • choose domain MHMICROSCOPY (not local computer e.g. Morpheus or Wheatstone)
    
     
• Start SimplePCI
  • run SimplePCI (C-Imaging)
    
    
    
     
  • During SimplePCI startup, 2 identical warning boxes will appear
    
  • Press OK or ignore messages concerning registry (report any other error messages to facility management and in the log book)
     
  • Main SimplePCI window will appear
    
     
• Start Capture mode in SimplePCI
  • Double click on camera icon
    
     
  • Two additional windows will open
    • The  capture box window (for camera control)
      
       
    • The Image Display window
      
       
• Ensure desired camera is chosen in top right selection box (01)
  • DVC-1412 on Morpheus
  • DVC-1312 on Wheatstone
    Change thumbnail to screen shot for 1412 or 1312)
     
• Choose number of color channels (02)
  • 
    

   

• Select proper Filter Setting for each color channel using the drop down menus
  • For previewing by eye, choose the WHITE LIGHT setting
  • For imaging a single fluorophore, choose WHITE LIGHT
  • For imaging multiple fluorophores using separate dichroic positions for each, choose manXXX settings for each fluorophore
    • ***Recommended for fixed samples
  • For imaging multiple fluorophores using a single dichroic for multiple wavelengths, choose the appropriate setting that does not have a man prefix
  • Click here for full description of all Filter Settings

   

• Ensure proper dichroic position is selected on the microscope stand!
   
• Press Focus button, which starts rapid acquisition of an image to the display window (03)
           
   
• Increase or decrease exposure time in order to get an image (03)
  • for fluorescence it may be desirable to increase camera gain to maximum (04) and decrease exposure time in order to make focusing easier.  Note that image may be noisy.  Black level should not be changed.
     
• Focus while viewing image on monitor
   
• For multi-color imaging: 
  Click on the "Red", "Green" or "Blue" tabs to change which channel is displayed in the Focus window
  NOTE:  The Focus image is always black and white
  •         [Insert Screen Image with tabs highlighted here]
     
• If desired, return camera gain to 41 (04) and increase exposure time (03) to reduce noise
   
• Press Stop (05) or Abort
           
   
• Press Capture 1 (06) to acquire - to ensure exposure is correct, check intensity histogram
  •         [Insert image of fluorescent sample here]
     
• Save the image
  • Right click on the image
  • Select "Save Image to File"
    • For Monochrome (single channel) images
      • ... "All Components" -- saves the image as monochrome
      • ... "Display Image" -- saves the image as index color
        • only available if you have changed the color display in the image window
    • For Multi-channel images
      • ... "All Components" -- saves all components to a single image, each channel is the color in which it was acquired
      • ... "Red/Green/Blue Component" -- saves a monochrome image of the color channel selected
      • ... "Display Image" -- saves a 24bit color image of what is displayed in the Image window