Sample image and/or code
open("D:/SnapMirror/ncc18jq_data/Research/Stem_Cell_Neural_Network/IPSC_images/CS20200610-well3/merged0717/20200717-20times-5_00219_Z001_Overlay.tif");
open("D:/SnapMirror/ncc18jq_data/Research/Stem_Cell_Neural_Network/IPSC_images/CS20200610-well3/merged0717/20200717-20times-5_00219_Z002_Overlay.tif");
open("D:/SnapMirror/ncc18jq_data/Research/Stem_Cell_Neural_Network/IPSC_images/CS20200610-well3/merged0717/20200717-20times-5_00219_Z003_Overlay.tif");
open("D:/SnapMirror/ncc18jq_data/Research/Stem_Cell_Neural_Network/IPSC_images/CS20200610-well3/merged0717/20200717-20times-5_00219_Z004_Overlay.tif");
open("D:/SnapMirror/ncc18jq_data/Research/Stem_Cell_Neural_Network/IPSC_images/CS20200610-well3/merged0717/20200717-20times-5_00219_Z005_Overlay.tif");
open("D:/SnapMirror/ncc18jq_data/Research/Stem_Cell_Neural_Network/IPSC_images/CS20200610-well3/merged0717/20200717-20times-5_00219_Z006_Overlay.tif");
open("D:/SnapMirror/ncc18jq_data/Research/Stem_Cell_Neural_Network/IPSC_images/CS20200610-well3/merged0717/20200717-20times-5_00219_Z007_Overlay.tif");
open("D:/SnapMirror/ncc18jq_data/Research/Stem_Cell_Neural_Network/IPSC_images/CS20200610-well3/merged0717/20200717-20times-5_00219_Z008_Overlay.tif");
run("Images to Stack", "name=0219_Overlay_Stack title=[] use");
run("Stack Focuser ", "enter=11");
close("0219_Overlay_Stack");
selectWindow("Focused_0219_Overlay_Stack");
// Color Thresholder 2.1.0/1.53c
// Autogenerated macro, single images only!
min=newArray(3);
max=newArray(3);
filter=newArray(3);
a=getTitle();
run("HSB Stack");
run("Convert Stack to Images");
selectWindow("Hue");
rename("0");
selectWindow("Saturation");
rename("1");
selectWindow("Brightness");
rename("2");
min[0]=126;
max[0]=129;
filter[0]="pass";
min[1]=1;
max[1]=223;
filter[1]="pass";
min[2]=230;
max[2]=255;
filter[2]="pass";
for (i=0;i<3;i++){
selectWindow(""+i);
setThreshold(min[i], max[i]);
run("Convert to Mask");
if (filter[i]=="stop") run("Invert");
}
imageCalculator("AND create", "0","1");
imageCalculator("AND create", "Result of 0","2");
for (i=0;i<3;i++){
selectWindow(""+i);
close();
}
selectWindow("Result of 0");
close();
selectWindow("Result of Result of 0");
rename(a);
// Colour Thresholding-------------
setOption("BlackBackground", true);
run("Convert to Mask");
run("Watershed");
run("Analyze Particles...", "size=50-Infinity pixel show=Outlines display summarize add");
Background
I am attempting to create a macro that can open 8 z stack images at a time, but I am unsure on how to automate the opening of these images. In my macro above, you can see that I am opening specific images, but I want to be able to generalize this. The images are all in the same file and are only organized by their file names (20200717-20times-5_00219_Z008_Overlay.tif). “219” is the specific image number and I want to be able to open all 8 images that contain that same image number.
Analysis goals
- What information are you interested in getting from this image? I am attempting to create a full focus image of immunostained neurons from 8 z stacks and then analyze the number of particles that fall within a certain color range.
Challenges
I’m not sure how to write the script to open 8 z stacked images simultaneously that have the same file name (the only part of the name that changes is Z001-Z008). I’m really new to this so I’m sorry if I’m not explaining my problem well!