CHAPTER 1 - SYSTEM OPERATIONZEISS Center Screen Area / Image Containers - Display and Image Analysis LSM 880474 000000-2071-464 10/2014 V_016.11.1 How a Scatter Diagram is GeneratedAll pixels having the same positions in both images are considered a pair. Of every pair of pixels (P1, P2)from the two source images, the intensity level of pixel P1 is interpreted as X coordinate, and that of pixelP2 as Y coordinate of the scatter diagram. The value of the pixel thus addressed is increased by one everytime, up to the maximum number of pixels used. This way, each pixel of the scatter diagram is a valuethat shows how often a particular pair of pixels has occurred.Differences between the images cause irregular spots in the scatter diagram.Identical images produce a clean diagonal line running from bottom left to top right, because only pixelpairs (0,0), (1,1), (2,2) with the same intensity can occur. Differences between the images cause anirregular distribution in the scatter diagram.Scatter diagram, image display and data table are interactively linked and immediately updatedwhen settings are changed.Available Controls:Drawing tools With the drawing tools the colocalization analysis can be restricted toa region of the image. Tools in the Colocalization View Option controlblock and in the Graphics View Option control block work the sameand can be combined.The Cross hair, Table and Image selection tick boxes: when selected,the respective element is displayed in the Image Display area. If off, theelement is hidden.Scaling pull down menu: Sets the histogram graph scaling to eitherdynamic (adjusting to the maximum value) or fixedSet the color LUTs for each quadrant in the image display to improvevisibility of colocalized pixels.The Load and Save buttons can be used to save colocalization settings(like color settings for the quadrants and crosshair-positions) to a *.colfile and re-load it when analyzing a different data set. This allowsusing the same parameter for the colocalization analysis of differentimages.The Cut Mask tool creates a new image document which sets everypixel outside Quadrant 3 (not colocalized) to Zero. ROIs don’t play arole for this Cut Mask function (as opposed to the Cut Mask in HistoView).Tables can always be saved or copied to the clipboard by right-mouse clicking on the tabledisplay!
