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Calculation for FRET Maps.
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macro "Ratiometric FRET" { | |
path = getDirectory("Choose a Directory"); | |
list = getFileList(path); | |
length = list.length; | |
File.makeDirectory(path + "Imaging/"); | |
//FRET map creation | |
for (i=0; i<length; i++) { | |
n=i+1; | |
open(path+list[i]); | |
img = File.nameWithoutExtension(); | |
selectWindow(img+".lsm"); | |
run("Subtract Background...", "rolling=300 stack"); | |
run("StackReg", "transformation=Translation"); | |
run("Median...", "radius=1.3 stack"); | |
run("Stack to Images"); | |
run("Duplicate...", "title=mask"); | |
run("Threshold..."); | |
setAutoThreshold("Default dark"); | |
waitForUser("Imagen "+n+" de "+length,"Apply Threshold y click OK"); | |
run("16-bit"); | |
run("Divide...", "value=255"); | |
saveAs("Tiff", path + "Imaging/mask 000" + i); | |
imageCalculator("Divide create 32-bit", "1-0002","1-0001"); | |
imageCalculator("Multiply create","Result of 1-0002","mask 000"+i+".tif"); | |
selectWindow("Result of Result of 1-0002"); | |
saveAs("Tiff", path + "Imaging/fret map " + i); | |
run("Close All"); | |
} | |
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