With reference to the image below: GFAP_Iba1-E84L1-20x-2 2 - Copy.tif (3.4 MB)
Could you please describe an optimal way to measure the fluorescence (mean grey area and SD)?
I.e: in the ROI circled below, what would be the best steps to quantify the fluorescence?
Currently the following steps are being undertaken: 1) subtract BG ; 2)convert to 8 bit 3)measure mean and SD 4)set threshold based on mean and SD. The mean values are perceived to be too low
GFAP_Iba1-E84L1-20x-2 2 - Copy - Copy.tif (3.8 MB)