Plant FP - please offer advice on what FP to choose

Probes & Prep
1

Good afternoon!

I am Francesco Cappai, a research at the University of Florida (US). My plant system exhibits very strong autofluorescence making GFP unusable. I will be ordering a synthetized a fluorescent-coding gene soon. I was looking at mTFP1 but I would love to hear your opinion about it.

If you had to try a fluorescent protein in a plant system with strong autofluorescence, what would you choose?

Thank you so much!

2

Hi Francesco -

I just imaged someones plant sample and they were using a RFP, I think it was DsRed. There was some dim autofluorescence, but not too bad on a 80 micro thick slice on a spinning disk.

3

Thank you! I’m a bit surprised though. From what I understood plant autofluorescence is strongest in the red side of the spectrum. I was not able to find a comprehensive review on the topic though.

4

Have you put your (unlabeled) sample on a microscope to see how strong the autofluorescence is with the different filter sets? That would tell you whether it’s bad or not in the red…

5

Thank you for your advice! I have a microscope that can go with gfp short and long pass, rfp and yfp. They all had bright autofluorescence. I don’t think the filter has any orange selectivity but I think I can have access to an instrument that does.