I have done immunofluorescence of tumour tissues for head and neck cancer to quantify the macrophages. What is the best method to quantify? I couldn't count manually as the cells are alot. What other techniques I can use?
Thank you.
Shams Aliya
This must be a merged image and the blue stain is DAPI. If so, on the splitted ones, calculate the integrative density of the stain (this can be done by ImageJ). Then, count the number of DAPI dots for the same slice. After that normalize the results by dividing the integrative density of the satin on the number of DAPI dots. Do this for about 5 to 15 image then plot them on a graph and perform the desired comparison.
Hi,
Can you explain a little bit more in detail what kind of measurements you want to perform?
What is a macrophage in that image and what is not?
Are just those cells with brightest particles in it?
How do you want to define a Macrophage?
Do you want to count a proportion of cells over the total number of cells?
Do you want to count the total number of cells in your image?
Automatic analysis goes after defining what do you want to measure/detect from the digital imaging point of view.
Explain a little bit more and we'll be happy to help,
Cheers,
J
J. Ramirez-Franco The macrophages is the green (around the nuclei), nuclei are blue. I think she want to count those cells.