I am attempting to visualize GFP expression in monocot pollen
transformed by microparticle bombardment. I have some questions:
1) Has anyone out there been able to increase the emission at 470 nm
excitation of the mGFP4 or mGFP5 (Haseloff) proteins by
photoconverting (bleaching) the 395 form to the 470 form? (refer to
Ultra-fast excited state dynamics in GFP paper by Chattoraj et al)
2) I have noticed a common filter used in fluorescence microscopy
detection of GFP is the LP520. If the GFP emission peak is 509, and
the LP520 blocks light below 520, why is it used? Why isn't a 510
narrow bandpass filter more common?
3) Does anyone have a transgenic plant (or seed) that expresses GFP
they can give to me? I would like to get my hands on some plant
cells that express GFP to calibrate my detection systems.
Crop Science, University of Guelph
acarlson at crop.uoguelph.ca