From the softWoRx main menu, choose Measure | Colocalization to open the Colocalization window.
in the Input field, enter the number of the Image window that you want to analyze.
To analyze a region of a window, select it as follows: Click Region and select an area in the Image window by dragging the mouse across the area. Adjust the rectangle you've created until it contains the desired area. Then click outside the Image window with the mouse.
Click Details to open the Region Details window. Specify the ranges of the X, Y , Z , and time data to analyze in the selected region.
In the Input Channels lists, select which channels to analyze.
Select a background threshold for each channel by clicking Get and then clicking on a background area of the image. The background is an averaged value within a box of the size specified.
When you are finished selecting the background, click the Get button again.
Click Do It to generate a product image, plot the Colocalization graph, and calculate the Pearson Coefficient of Correlation (this value is displayed in the Colocalization window).