I'm currently doing research on the reduction of contaminates on dog collars, using GloGerm as my pseudo contaminate. Originally, I was using a scoring system to rank the amount of reduction but quickly discovered that having a value that labeled the total amount of reduction would further solidify my results. I'm not sure if the way I'm measuring fluorescence through ImageJ is correct or not. I found a link to instructions that someone used to determine the Corrected Total Cell Fluorescence (CTCF). See link below.
I'm not sure if this is the correct way to go about measuring my reduction of fluorescence. Currently, I'm having issues with my pre and post images. I'm getting values that state that the post image shows greater fluorescence compared to the pre. Which, suggests the addition of contaminate. However, I'm washing the the collars. So post should indicate some sort of reduction.
Here is a sample of the pictures that I'm using. I'm not sure if the link to that article is the best source or not to use to determine the amount of reduction. Is it incorrect to use that format for analysis or do you suggest a different method to find those measurements? I'm not measuring a cell's fluorescence, so I'm not sure if ImageJ will work for my type of analysis. Any help and suggestions you could provide would be greatly appreciated.