// Calcein stim figures images

tstack = getTitle(); 

run("Z Project...", "stop=30 projection=[Average Intensity]");
avgprj = getTitle(); 
run("Enhance Contrast", "saturated=0.35");

selectWindow(tstack);

frame = 135;
run("Duplicate...", "duplicate range=&frame-&frame");
init = getTitle(); 

selectWindow(tstack);

frame = 145;
run("Duplicate...", "duplicate range=&frame-&frame");
diff = getTitle(); 

imageCalculator("Subtract create 32-bit", diff, init);
num = getTitle(); 

imageCalculator("Divide create 32-bit", num, init);
setMinAndMax("-0.1", "0.10");
run("!WRA_OrangeLowCyanHigh");
diffimg = getTitle();

close(init);
close(diff); 
close(num);