Batch processing timepoint fluorescence images

Hey Everyone,

I’m a newbie and not a programmer so working with macros is pretty tough for me. In a cell experiment measured the fluorescence of 6 groups of cells t DAPI, 647, 568 and 488. I imaged these over 11 timepoints. In summary, I took images of 12 groups at timepoints every 3 mins for about 30 mins using 4 different channels. What I want to do is apply the same processing to each group for all timepoints (ie brightness correction etc in each channel), then quantify for each group/ timepoint the 488 fluorescence intensities in the ROIs and compare between timepoints (this should be possible as we use the same acquisition parameters). Since I need the processing to be the same for this analysis I guess I need some macro script to do it for me.

Would anyone be able to help me!

Thanks alot!

Hi @Chloe2, and welcome to the forum!

don’t be afraid of macros, you can use the macro recorder to get what you need on one image, and then apply it to multiple images as documented here:

Let us know if you get stuck, and feel free to post more specific follow-up questions here.

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