Hi all, thank you for your lending time and your experience to my question!
I understand phenol red is discouraged when pursuing live imaging because it interferes with the fluorescence. However, IF-smFISH is not a live cell imaging process: cells are washed twice with 1x PBS prior to fixation then washed three more times with 1x PBS prior to being blocked. Because of this, I think that we should be able to use phenol red in our media so long as it’s not taken up by the neurons during culture. Does anyone have any insight on this?
You are correct. If you are not imaging the cells within phenol-red containing media, you should be totally fine. It is just live-imaging, during media that contains some fluorescent properties that is a concern in this case. Good luck!
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