How A Scatter Diagram Is Generated - Zeiss LSM 880 Operating Manual

ZEISS Center Screen Area / Image Containers - Display and Image Analysis
6.11.1

How a Scatter Diagram is Generated

All 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 pixel
P2 as Y coordinate of the scatter diagram. The value of the pixel thus addressed is increased by one every
time, up to the maximum number of pixels used. This way, each pixel of the scatter diagram is a value
that 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 pixel
pairs (0,0), (1,1), (2,2) with the same intensity can occur. Differences between the images cause an
irregular distribution in the scatter diagram.
Scatter diagram, image display and data table are interactively linked and immediately updated
when settings are changed.
Available Controls:
Drawing tools
Tables can always be saved or copied to the clipboard by right-mouse clicking on the table
display!
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CHAPTER 1 - SYSTEM OPERATION
With the drawing tools the colocalization analysis can be restricted to
a region of the image. Tools in the Colocalization View Option control
block and in the Graphics View Option control block work the same
and 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, the
element is hidden.
Scaling pull down menu: Sets the histogram graph scaling to either
dynamic (adjusting to the maximum value) or fixed
Set the color LUTs for each quadrant in the image display to improve
visibility 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 *.col
file and re-load it when analyzing a different data set. This allows
using the same parameter for the colocalization analysis of different
images.
The Cut Mask tool creates a new image document which sets every
pixel outside Quadrant 3 (not colocalized) to Zero. ROIs don't play a
role for this Cut Mask function (as opposed to the Cut Mask in Histo
View).
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LSM 880
10/2014 V_01