[Histonet] fluorescent proteins (RFP and GFP) native fluorescence
I was hoping someone here could give me some advice on their
experience with various fluorescent proteins. I need a good marker
for my viral vector. I have a humanized GFP, RFP and EGFP. I would
like to inject the vector into the tail vein to see which tissue
lights up. My hGFP variant is not very strong in vitro, but both RFP
and EGFP work well. I have heard that GFP quickly disperses when
mounted on slides. However, I have seen some papers that use native
EGFP fluorescence with cryosections and they don't have problems.
Ideally I would use floating tissue sections of 40 microns as I have
easy access to a sliding microtome. I have heard that RFP does not
work with fixation at all.
I am open to any sort of advice. Could someone recommend a protocol
for visualizing native fluorescence with either EGFP or RFP?
Specifically, whether to fix or not, the thickness of the section,
floating sections vs. slide mounts, etc.
Also, if there is another fluorescent protein available that you
could suggest I would like to hear about it. I have heard of yellow
and blue fluorescent proteins as well as a monomeric form or RFP. I
am also open to using a "universal" fusion protein, for example actin-
GFP if it solves the problem of GFP native fluorescence.
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