Can anybody tell me why with this macro I just get as an output for each channel of each image a .png file of the first image slice and not of the stacked image? It seems like the Z-project part of the code is not properly working, thanks in advance!!
`// Function to process each .nd2 file
function processFile(filePath) {
// Open the .nd2 file
setBatchMode(true);
open(filePath);
// Get the title of the opened image
title = getTitle();
// Stack different focal planes into one high projection image per channel
run("Split Channels");
channelTitles = getList("image.titles");
// Process each channel
for (c = 0; c < channelTitles.length; c++) {
selectWindow(channelTitles[c]);
// Get the total number of slices
totalSlices = getSliceNumber();
// Use all slices for Z Projection
run("Z Project...", start=1 stop=totalSlices, " projection=[Max Intensity]");
run("Update");
projTitle = getTitle();
close(channelTitles[c]);
selectWindow(projTitle);
// Adjust brightness and contrast automatically
run("Enhance Contrast", "saturated=0.35");
run("Update")
run("Apply LUT", "stack");
// Convert to RGB
run("RGB Color");
// Save the processed channel
saveAs("Tiff", filePath + "_channel" + (c + 1) + ".tif");
close();
}
// Close the original image
close(title);
}`
I tried to work on the following portion of the code by selecting the project image once generated and closing the original one but it doesn't still work....
// Use all slices for Z Projection
run("Z Project...", start=1 stop=totalSlices, " projection=[Max Intensity]");
run("Update");
projTitle = getTitle();
close(channelTitles[c]);
selectWindow(projTitle);
That's what I get after all the attempt and suggestions you gave me
Below please find an ImageJ demo macro that uses a ".nd2"-file I found here. Please download this raw-file and try the macro with this file! The file shows two colour-channels with four images (here: time points).
//imagej-macro "nd2-processing" (Herbie G., 09. Aug. 2024)
requires("1.54j");
path=File.openDialog("Open the sample nd2-file");
run("Bio-Formats","open=[path] color_mode=Default rois_import=[ROI manager] split_channels view=Hyperstack stack_order=XYCZT use_virtual_stack series_1");
path=File.getDirectory(path)+File.nameWithoutExtension;
clr=newArray("Blue","Green","Red");
n=nImages;
for (i=0;i<n;i++) {
run("Z Project...","projection=[Max Intensity]");
run("Enhance Contrast...","saturated=0.35");
run(clr[i]);
saveAs("PNG",path+"_ch-"+(n-i)+".png");
close(); close();
}
exit();
//imagej-macro "nd2-processing" (Herbie G., 09. Aug. 2024)
Please note that by assigning a LUT to the projection images makes them saved with 8bit depth. Save in TIF-format to keep the 16bit depth. =============================================
Finally, below please find the code for batch processing all ".nd2"-files in a folder:
//imagej-macro "batch_nd2-processing" (Herbie G., 09./10. Aug. 2024)
requires("1.54j");
close("*");
inSfx=2; // preferred source file suffix
outSfx=0; // preferred result file suffix
Dialog.create("Batch Process Folder");
sfxIn=newArray(".tif",".czi",".nd2");
sfxOut=newArray(".tif",".png",".jpg");
Dialog.setInsets(0,220,0);
Dialog.addMessage("Macro does not process nested folders!",12,"blue");
Dialog.setInsets(6,0,0);
Dialog.addChoice("Suffix",sfxIn,sfxIn[inSfx]);
Dialog.setInsets(-28,182,0);
Dialog.addMessage("of the source files");
Dialog.addDirectory("Source-Folder",getDir("file"));
Dialog.addChoice("Suffix",sfxOut,sfxOut[outSfx]);
Dialog.setInsets(-37,182,0);
Dialog.addMessage("for the result files");
Dialog.addDirectory("Result-Folder",getDir("file"));
Dialog.show();
sfx1=Dialog.getChoice();
srce=Dialog.getString();
sfx2=Dialog.getChoice();
dest=Dialog.getString();
setBatchMode(true);
count=processFolder(sfx1,srce,sfx2,dest);
setBatchMode(false);
exit(""+count+" files processed");
//
function processFolder(xtn1,dir1,xtn2,dir2) {
color=newArray("Blue","Green","Red","Yellow","Magenta","Cyan");
cnt=0;
list=getFileList(dir1);
n=list.length;
for (i=0;i<n;i++)
if (endsWith(list[i],xtn1)) {
path=dir1+list[i];
run("Bio-Formats","open=[path] color_mode=Default split_channels view=Hyperstack stack_order=XYCZT use_virtual_stack series_1");
path=dir2+File.nameWithoutExtension;
Stack.getDimensions(w,h,c,s,f);
if (s>1||f>1) {
cnt++;
processStack(path,xtn2,color);
} else {
close("*");
print("Unsuited image data: "+list[i]);
}
}
return cnt;
}
function processStack(pth,xtn,clr) {
n=nImages;
for (i=0;i<n;i++) {
run("Z Project...","projection=[Max Intensity]");
run("Enhance Contrast...","saturated=0.35");
if (n<7) run(clr[i]);
saveAs(pth+"_ch-"+(n-i)+xtn);
close(); close();
}
}
//imagej-macro "batch_nd2-processing" (Herbie G., 09./10. Aug. 2024)
It is assumed that the source stacks have a maximum of six colour channels!