Hello everyone,
I've stained for astrocytes in the human brainstem. Attached is an image I took from the fluorescence microscope. The cell density is too high, I'm wondering whether it'll be possible to count the number of astrocytes using Fiji/ImageJ.
Based on this image, no. However, if you stain for nuclei and then go the particle or cell counter, you can then manually select each of the nuclei and imageJ will keep track of them for you. You can also track different cell types in the same image provided you can tell them apart, You need a nuclei stain here to determine how many cells you have.
For real cell counting, you should do a double label with an astrocyte marker and DAPI for the nucleus. Then count the number of nucleus having the astrocyte staining. ImageJ should easily be able to count the DAPI stained nucleus.
On second thought, you don't really need 'cell counting', the total fluorescence in the image (number of pixels x grey values) should roughly correspond to the number of cells. It will not be an exact match with the cell number, but it will give you a very good estimation and you can count very easily a large number of samples. For comparing between images, you will need to do 'a normalization' of the grey scale value. That is ((astrocyte florescence intensity-background)/astrocyte fluorescence intensity), for making the fluorescence intensity equal between the images. That way you can quantify the image.
Regards
- Badges
- Science method
- Similar topics
- Biological Science
- Histology
- Staining