Suggestion required_please help

imagej
segmentation
microscopy

#1

Hi everybody,

I am new to Image J, but a quick learner if you can just suggest me few things. So in the attached image you can see there is an image which was taken confocal microscopy and if you look closely, you see lots of circular droplets of fat.

Now I wish to quantify them for their diameter and I wish to find their number and diameter so that I can perform some statistical analysis.

Just suggest me how to proceed?
Which software or toolbox or part of Image J(or other software like it), should I use?
Can it be done in one go which yield a table of output containing diameter / volume or may be location.

Please just help me with little detail and I will fly.
Thanks in advance.

Nikita


#2

No worries @Nikita1 … Let’s get you started with some SUPER helpful links and resources:

Going through the Introduction and Segmentation workshops will be really helpful I should think… :slight_smile: What you need to do is segment those lipid droplets… MorphoLibJ is a great tool for such things… :slight_smile: For example… I converted your image to 8-bit (though ideally - you have the original files in the file formats in which they were acquired on your system - NOT jpegs!), Enhance Contrast... using default settings, and then used MorphoLibJ to get this result (with these settings):

For sure it’s not perfect… then you’ll have to select objects that are more circular, but varying size. But that’s a start anyway. :slight_smile:

Go through those links above - take the time to do that… and if you have more specific questions down-the-road… post again.

eta :slight_smile: