I am postdoc who study the odor response of insect antennal lobe.
I use calcium imaging to observe which part of the antennal lobe could be activated by a certain odorant. I choose calcium green which activated by 475nm light as the indicator. Calcium green face to a strong bleaching thus I need to do bleach correction.
Current I use fiji.
Image>adjust>bleach correction–Exponential Fit.
Then I calculate the △F/F0 to know the intensity of the frame.
I read the instruction of bleach correction–exponential fit, it is said that “since bleaching is often not mono-exponential, quantification of fluorescence intensities after bleach correction is not possible. This plugin should only be used to enhance time-course movies for presentation rather than quantification”.
I want to know how to analysis the data when I want to quantify the intensity of the fluorescence also need to do the bleach correction? It seems that bleach correction might bring errors into the data.
Thank you for reading my question.