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fixed-images-general-220608-b1int-IF-time-course-get-arguments.ijm
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fixed-images-general-220608-b1int-IF-time-course-get-arguments.ijm
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inputFolder = getArgument();
// Create an output folder based on the inputFolder
parentFolder = getPath(inputFolder); inputFolderPrefix = getPathFilenamePrefix(inputFolder);
outputFolder = parentFolder + inputFolderPrefix + "-output" + File.separator;
if ( !(File.exists(outputFolder)) ) { File.makeDirectory(outputFolder); }
run("Close All");
setBatchMode(true);
//c1name = "DAPI";
//c2name = "Col4a1-TMR";
c3name = "NLSmSL";
c4name = "b1int";
type = "Specify Slice Number";
specifiedSliceNumber = 1;
keepTemp = "Yes";
processIFfolder(inputFolder, outputFolder);
function processSingleSlice(id, typePrefix, outputFolder, outputPrefix) {
// idC1 = getChannel(id, 1);
// idC2 = getChannel(id, 2);
idC3 = getChannel(id, 3);
idC4 = getChannel(id, 4);
// saturation = 0.1; idC1_8bit = to8bitSatu( idC1, typePrefix + "-" + c1name, saturation, outputFolder, outputPrefix );
// c1min = 70; c1max = 7000; idC1_8bit = to8bitMinMax( idC1, typePrefix + "-" + c1name, c1min, c1max, outputFolder, outputPrefix );
// saturation = 0.1; idC2_8bit = to8bitSatu( idC2, typePrefix + "-" + c2name, saturation, outputFolder, outputPrefix );
// c2min = 0; c2max = 800; idC2_8bit = to8bitMinMax( idC2, typePrefix + "-" + c2name, c2min, c2max, outputFolder, outputPrefix );
// saturation = 0.5; idC3_8bit = to8bitSatu( idC3, typePrefix + "-" + c3name, saturation, outputFolder, outputPrefix );
saturation = 0.1; gamma_value = 0.5; idC3_8bit = to8bitSatuGamma( idC3, typePrefix + "-" + c3name, saturation, gamma_value, outputFolder, outputPrefix );
// c3min = 0; c3max = 4000; idC3_8bit = to8bitMinMax( idC3, typePrefix + "-" + c3name, c3min, c3max, outputFolder, outputPrefix );
// saturation = 0.2; idC4_8bit = to8bitSatu( idC4, typePrefix + "-" + c4name, saturation, outputFolder, outputPrefix );
c4min = 0; c4max = 1000; idC4_8bit = to8bitMinMax( idC4, typePrefix + "-" + c4name, c4min, c4max, outputFolder, outputPrefix );
mergeGM( idC4_8bit, c4name, idC3_8bit, c3name, outputFolder, outputPrefix );
// mergeBGMY( idC1_8bit, c1name, idC2_8bit, c2name, idC3_8bit, c3name, idC4_8bit, c4name, outputFolder, outputPrefix );
// mergeBGY( idC1_8bit, c1name, idC2_8bit, c2name, idC3_8bit, c3name, outputFolder, outputPrefix );
// mergeBGM( idC1_8bit, c1name, idC2_8bit, c2name, idC4_8bit, c4name, outputFolder, outputPrefix );
// mergeBGM( idC1_8bit, c1name, idC3_8bit, c3name, idC2_8bit, c2name, outputFolder, outputPrefix );
return 1;
}
function processIFfolder(inputFolder, outputFolder) {
tempFolder = outputFolder + inputFolderPrefix + "-temp" + File.separator;
if ( !(File.exists(tempFolder)) ) { File.makeDirectory(tempFolder); }
flist = getFileList(inputFolder);
//for (i=0; i<1; i++) {//for testing
for (i=0; i<flist.length; i++) {
filename = inputFolder + flist[i];
outputPrefix = getFilenamePrefix(flist[i]);
if ( endsWith(filename, ".nd2") || endsWith(filename, ".czi") || endsWith(filename, ".tif") ) {
if ( type == "Max Intensity Projection" ) {
tempFile = tempFolder + outputPrefix + "-MIP" + ".tif";
if ( File.exists(tempFile) ) { open(tempFile); idMIP = getImageID(); }
else { open(filename); id0 = getImageID(); idMIP = getMIP(id0, tempFile); }
processSingleSlice(idMIP, "MIP", outputFolder, outputPrefix);
}
if ( type == "Middle Slice" ) {
tempFile = tempFolder + outputPrefix + "-midZ" + ".tif";
if ( File.exists(tempFile) ) { open(tempFile); idMidZ = getImageID(); }
else { open(filename); id0 = getImageID(); idMidZ = getMidZ(id0, tempFile); }
processSingleSlice(idMidZ, "midZ", outputFolder, outputPrefix);
}
if ( type == "Specify Slice Number" ) {
tempFile = tempFolder + outputPrefix + "-z-"+specifiedSliceNumber + ".tif";
if ( File.exists(tempFile) ) { open(tempFile); idZ = getImageID(); }
else { open(filename); id0 = getImageID(); idZ = getZslice(id0, specifiedSliceNumber, tempFile); }
processSingleSlice(idZ, "z-"+specifiedSliceNumber, outputFolder, outputPrefix);
}
run("Close All");
}
}
if ( keepTemp == "No" ) deleteFolder(tempFolder);
}
function getChannel(id, c) {
selectImage(id); run("Duplicate...", "duplicate channels="+c); idC = getImageID(); return idC;
}
function getMidZ(id, tempFile) {
selectImage(id);
if ( nSlices > 1 ) { Stack.getDimensions(w, h, c, s, f); } //width, height, channels, slices, frames
if ( f == 1 ) { idf1 = id; }
if ( f > 1 && c*s == 1 ) {
// when only f > 1, in duplicate it has to be called "range"
selectImage(id); run("Duplicate...", "duplicate range=1-1"); idf1 = getImageID();
print("There are multiple time frames. Processing only the first time point.");
print("WARNING: It is unusual to have multiple frames for IF images, please look into your images.");
}
if ( f > 1 && c*s > 1 ) {
selectImage(id); run("Duplicate...", "duplicate frames=1"); idf1 = getImageID();
print("There are multiple time frames. Processing only the first time point.");
print("WARNING: It is unusual to have multiple frames for IF images, please look into your images.");
}
if ( s == 1 ) { selectImage(idf1); save(tempFile); return idf1; }
if ( s > 1 ) {
sMid = floor( s/2 );
// when only f > 1, in duplicate it has to be called "range"
if ( c == 1 ) { selectImage(idf1); run("Duplicate...", "duplicate range="+sMid+"-"+sMid); idMidZ = getImageID(); }
if ( c > 1 ) { selectImage(idf1); run("Duplicate...", "duplicate slices="+sMid); idMidZ = getImageID(); }
selectImage(idMidZ); save(tempFile); return idMidZ;
}
}
function getZslice(id, zN, tempFile) {
selectImage(id);
if ( nSlices > 1 ) { Stack.getDimensions(w, h, c, s, f); } //width, height, channels, slices, frames
if ( f == 1 ) { idf1 = id; }
if ( f > 1 && c*s == 1 ) {
// when only f > 1, in duplicate it has to be called "range"
selectImage(id); run("Duplicate...", "duplicate range=1-1"); idf1 = getImageID();
print("There are multiple time frames. Processing only the first time point.");
print("WARNING: It is unusual to have multiple frames for IF images, please look into your images.");
}
if ( f > 1 && c*s > 1 ) {
selectImage(id); run("Duplicate...", "duplicate frames=1"); idf1 = getImageID();
print("There are multiple time frames. Processing only the first time point.");
print("WARNING: It is unusual to have multiple frames for IF images, please look into your images.");
}
if ( s == 1 ) { selectImage(idf1); save(tempFile); return idf1; }
if ( s > 1 ) {
// when only f > 1, in duplicate it has to be called "range"
if ( c == 1 ) { selectImage(idf1); run("Duplicate...", "duplicate range="+zN+"-"+zN); idzN = getImageID(); }
if ( c > 1 ) { selectImage(idf1); run("Duplicate...", "duplicate slices="+zN); idzN = getImageID(); }
selectImage(idzN); save(tempFile); return idzN;
}
}
function getMIP(id, tempFile) {
selectImage(id);
//if ( nSlices > 1 ) { Stack.getDimensions(w, h, c, s, f); } //width, height, channels, slices, frames
Stack.getDimensions(w, h, c, s, f); //width, height, channels, slices, frames
if ( f == 1 ) { idf1 = id; }
if ( f > 1 && c*s == 1 ) {
// when only f > 1, in duplicate it has to be called "range"
selectImage(id); run("Duplicate...", "duplicate range=1-1"); idf1 = getImageID();
print("There are multiple time frames. Processing only the first time point.");
print("WARNING: It is unusual to have multiple frames for IF images, please look into your images.");
}
if ( f > 1 && c*s > 1 ) {
selectImage(id); run("Duplicate...", "duplicate frames=1"); idf1 = getImageID();
print("There are multiple time frames. Processing only the first time point.");
print("WARNING: It is unusual to have multiple frames for IF images, please look into your images.");
}
if ( s == 1 ) { selectImage(idf1); save(tempFile); return idf1; }
if ( s > 1 ) { selectImage(idf1); run("Z Project...", "projection=[Max Intensity]"); idMIP = getImageID(); save(tempFile); return idMIP;}
}
function makeMIPfolder(inputFolder, outputFolder) {
flist = getFileList(inputFolder);
for (i=0; i<flist.length; i++) {
filename = inputFolder + flist[i];
outputPrefix = getFilenamePrefix(flist[i]);
if ( endsWith(filename, ".nd2") || endsWith(filename, ".czi") || endsWith(filename, ".tif") ) {
open(filename); id0 = getImageID();
idMIP = getMIP(id0);
save(outputFolder + outputPrefix + "-MIP.tif");
run("Close All");
}
}
// scale the image using saturation, change to 8-bit and save it
function to8bitSatuGamma( id, cName, satu, gamma_value, outputFolder, outputPrefix ){
selectImage(id); run("Grays"); run("Gamma...", "value="+gamma_value); run("Enhance Contrast", "saturated="+satu); run("8-bit");
outputFilename = outputPrefix + "-" + cName + "-8bit.tif";
saveAs("Tiff", outputFolder + outputFilename);
return outputFilename;
//id8bit = getImageID(); return id8bit;
}
// scale the image using saturation, change to 8-bit and save it
function to8bitSatu( id, cName, satu, outputFolder, outputPrefix ){
selectImage(id); run("Grays"); run("Enhance Contrast", "saturated="+satu); run("8-bit");
outputFilename = outputPrefix + "-" + cName + "-8bit.tif";
saveAs("Tiff", outputFolder + outputFilename);
return outputFilename;
//id8bit = getImageID(); return id8bit;
}
// scale the image using specified min and max, change to 8-bit and save it
function to8bitMinMax( id, cName, imgMin, imgMax, outputFolder, outputPrefix ){
selectImage(id); run("Grays"); setMinAndMax(imgMin, imgMax); run("8-bit");
outputFilename = outputPrefix + "-" + cName + "-8bit.tif";
saveAs("Tiff", outputFolder + outputFilename);
return outputFilename;
//id8bit = getImageID(); return id8bit;
}
// open 2 images, make a 2 x 1 montage from them
function montage2( f1, f2, outputFolder, outputPrefix ){
open(outputFolder + f1); rename("montage-1");
open(outputFolder + f2); rename("montage-2");
run("Images to Stack", "name=Stack title=[montage] use");
run("Make Montage...", "columns=2 rows=1 scale=0.50");
outputFilename = outputPrefix + "-2Montage.tif";
saveAs("Tiff", outputFolder + outputFilename);
return outputFilename;
}
// open 3 images, make a 3 x 1 montage from them
function montage3( f1, f2, f3, outputFolder, outputPrefix ){
open(outputFolder + f1); rename("montage-1");
open(outputFolder + f2); rename("montage-2");
open(outputFolder + f3); rename("montage-3");
run("Images to Stack", "name=Stack title=[montage] use");
run("Make Montage...", "columns=3 rows=1 scale=0.50");
outputFilename = outputPrefix + "-3Montage.tif";
saveAs("Tiff", outputFolder + outputFilename);
return outputFilename;
}
function montage4( f1, f2, f3, f4, outputFolder, outputPrefix ){
// open 4 images, make a 4 x 1 montage from them
open(outputFolder + f1); rename("montage-1");
open(outputFolder + f2); rename("montage-2");
open(outputFolder + f3); rename("montage-3");
open(outputFolder + f4); rename("montage-4");
run("Images to Stack", "name=Stack title=[montage] use");
run("Make Montage...", "columns=4 rows=1 scale=0.50");
outputFilename = outputPrefix + "-4Montage.tif";
saveAs("Tiff", outputFolder + outputFilename);
return outputFilename;
}
function mergeGM( cGreen, greenName, cMagenta, magentaName, outputFolder, outputPrefix ){
// merge channels in order in green and magenta colors
// c1: red; c2: green; c3:blue; c4:gray; c5:cyan; c6: magenta; c7: yellow
run("Merge Channels...", "c2=["+cGreen+"] c6=["+cMagenta+"] keep");
outputFilename = outputPrefix + "-" + greenName + "_inGreen-" + magentaName + "_inMagenta.tif";
saveAs("Tiff", outputFolder + outputFilename);
return outputFilename;
}
function mergeGC( cGreen, greenName, cCyan, cyanName, outputFolder, outputPrefix ){
// merge channels in order in green and magenta colors
// c1: red; c2: green; c3:blue; c4:gray; c5:cyan; c6: magenta; c7: yellow
run("Merge Channels...", "c2=["+cGreen+"] c5=["+cCyan+"] keep");
outputFilename = outputPrefix + "-" + greenName + "_inGreen-" + cyanName + "_inCyan.tif";
saveAs("Tiff", outputFolder + outputFilename);
return outputFilename;
}
function mergeCM( cCyan, cyanName, cMagenta, magentaName, outputFolder, outputPrefix ){
// merge channels in order in green and magenta colors
// c1: red; c2: green; c3:blue; c4:gray; c5:cyan; c6: magenta; c7: yellow
run("Merge Channels...", "c5=["+cCyan+"] c6=["+cMagenta+"] keep");
outputFilename = outputPrefix + "-" + cyanName + "_inCyan-" + magentaName + "_inMagenta.tif";
saveAs("Tiff", outputFolder + outputFilename);
return outputFilename;
}
function mergeBGr( cBlue, blueName, cGray, grayName, outputFolder, outputPrefix ){
// merge channels in order in blue and gray colors
// c1: red; c2: green; c3:blue; c4:gray; c5:cyan; c6: magenta; c7: yellow
run("Merge Channels...", "c3=["+cBlue+"] c4=["+cGray+"] keep");
outputFilename = outputPrefix + "-" + blueName + "_inBlue-" + grayName + "_inGray.tif";
saveAs("Tiff", outputFolder + outputFilename);
return outputFilename;
}
function mergeBGM( cBlue, blueName, cGreen, greenName, cMagenta, magentaName, outputFolder, outputPrefix ){
// merge channels in order in blue, green and magenta colors
// c1: red; c2: green; c3:blue; c4:gray; c5:cyan; c6: magenta; c7: yellow
run("Merge Channels...", "c2=["+cGreen+"] c3=["+cBlue+"] c6=["+cMagenta+"] keep");
outputFilename = outputPrefix + "-" + blueName + "_inBlue-" + greenName + "_inGreen-" + magentaName + "_inMagenta.tif";
saveAs("Tiff", outputFolder + outputFilename);
return outputFilename;
}
function mergeBGY( cBlue, blueName, cGreen, greenName, cYellow, yellowName, outputFolder, outputPrefix ){
// merge channels in order in blue, green and magenta colors
// c1: red; c2: green; c3:blue; c4:gray; c5:cyan; c6: magenta; c7: yellow
run("Merge Channels...", "c2=["+cGreen+"] c3=["+cBlue+"] c7=["+cYellow+"] keep");
outputFilename = outputPrefix + "-" + blueName + "_inBlue-" + greenName + "_inGreen-" + yellowName + "_inYellow" + ".tif";
saveAs("Tiff", outputFolder + outputFilename);
return outputFilename;
}
function mergeGMY( cGreen, greenName, cMagenta, magentaName, cYellow, yellowName, outputFolder, outputPrefix ){
// merge channels in order in blue, green and magenta colors
// c1: red; c2: green; c3:blue; c4:gray; c5:cyan; c6: magenta; c7: yellow
run("Merge Channels...", "c2=["+cGreen+"] c6=["+cMagenta+"] c7=["+cYellow+"] keep");
outputFilename = outputPrefix + "-" + greenName + "_inGreen-" + magentaName + "_inMagenta-" + yellowName + "_inYellow" + ".tif";
saveAs("Tiff", outputFolder + outputFilename);
return outputFilename;
}
function mergeCGMY( cCyan, cyanName, cGreen, greenName, cMagenta, magentaName, cYellow, yellowName, outputFolder, outputPrefix ){
// merge channels in order in blue, green and magenta colors
// c1: red; c2: green; c3:blue; c4:gray; c5:cyan; c6: magenta; c7: yellow
run("Merge Channels...", "c2=["+cGreen+"] c5=["+cCyan+"] c6=["+cMagenta+"] c7=["+cYellow+"] keep");
outputFilename = outputPrefix + "-" + cyanName + "_inCyan-" + greenName + "_inGreen-" + magentaName + "_inMagenta-" + yellowName + "_inYellow.tif";
saveAs("Tiff", outputFolder + outputFilename);
return outputFilename;
}
function mergeBGMY( cBlue, blueName, cGreen, greenName, cMagenta, magentaName, cYellow, yellowName, outputFolder, outputPrefix ){
// merge channels in order in blue, green and magenta colors
// c1: red; c2: green; c3:blue; c4:gray; c5:cyan; c6: magenta; c7: yellow
run("Merge Channels...", "c2=["+cGreen+"] c3=["+cBlue+"] c6=["+cMagenta+"] c7=["+cYellow+"] keep");
outputFilename = outputPrefix + "-" + blueName + "_inBlue-" + greenName + "_inGreen-" + magentaName + "_inMagenta-" + yellowName + "_inYellow.tif";
saveAs("Tiff", outputFolder + outputFilename);
return outputFilename;
}
function getPath(pathFileOrFolder) {
// this one takes full path of the file (input can also be a folder) and returns the parent folder path
temp = split(pathFileOrFolder, File.separator);
if ( File.separator == "/" ) {
// Mac and unix system
pathTemp = File.separator;
for (i=0; i<temp.length-1; i++) {pathTemp = pathTemp + temp[i] + File.separator;}
}
if ( File.separator == "\\" ) {
// Windows system
pathTemp = temp[0] + File.separator;
for (i=1; i<temp.length-1; i++) {pathTemp = pathTemp + temp[i] + File.separator;}
}
return pathTemp;
}
function getPathFilenamePrefix(pathFileOrFolder) {
// this one takes full path of the file
temp = split(pathFileOrFolder, File.separator);
temp = temp[temp.length-1];
temp = split(temp, ".");
return temp[0];
}
function getFilenamePrefix(filename) {
// this one takes just the file name without folder path
temp = split(filename, ".");
return temp[0];
}
function deleteFolder(folder) {
// Delete all the files inside the folder, then the folder itself
list = getFileList(folder);
// Delete the files and the folder
for (i=0; i<list.length; i++){
ok = File.delete(folder+list[i]);
}
ok = File.delete(folder);
if (File.exists(folder))
exit("Unable to delete the folder: " + folder);
else
print("Successfully deleted: " + folder);
}