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Recently a new faculty member has introduced new constructs
of both YFP and CFP for localization in plant cells, mostly tobacco and
arabidopsis. Our current system, a FluoView 500 is not set up for this
work, so I've been asked to investigate new avenues.
In any case, I think we've all seen a spectral system separate GFP, Sytox Green
and FITC in the same cell, but for CFP, YFP, GFP, RFP colocalization in plant
cells would a spectral system offer more utility than a system with better
filter sets and laser lines?
Obviously there is a cost question in relation to utility here. Also,
here in Midwest of the US, we have primarily Zeiss, Olympus and Nikon
systems. If anyone would like comment on those in particular, that'd be
great.
I also have a more specific question, with several groups targeting
chloroplasts, I should probably just lean towards a spectral system?
Finally, do YFP and GFP overlap too greatly in plants (pH difference?!?) to be
separated by either system?
If any has some negative feedback, or suggestions, please feel free to email me
privately. I find folks are awfully nice on the list, but when we're
talking several hundred thousand dollars, I need brutal honesty.
Thank you.
Christian
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