I took my pictures with a confocal microscope. I would like to measure the whole picture fluorescence intensity and calculate the cell count. I have a nuclear staining with DAPI, actin filaments with Phalloidin and E-Cadherin with Alexa488. I don't know what could be the best way to measure the fluorescence. Maybe if I split the channels, then adjust the threshold in every channel seperately and then measure the thresholded area intensity? Or If I select a ROI and then measure the intensity? Help me please... Thank you!
Loretta
Hi,
I would split channels, convert everything to b/w 8bit and then measure ROIs manually. You will get mean grey values of your measurement and will have to normalize it anyhow?
Best,
Kustrim
Hi Kustrim Cerimi !
I read that it is better to measure the integrated density, and I would like to measure the whole picture fluorescence intensity, because I would like to compare how the treatment reduced or increased the red and green intensities.
What are you exactly measuring? Cells? Cropped cell images? If your imaging different cells I would suggest to firstly threshold all the background out and then to measure tho overall intensity.
- Badges
- Science topic
- Similar topics
- Physical Sciences
- Physical Phenomena
- Luminescence
- Fluorescence