The highlight of day two of ABRF, for me, was the talk by Robert Campbell on fluorescent reporters. He started off by saying that existing fluorescent proteins were quite good and unlikely to get dramatically better. He spent the rest of his talk discussing his labs work to improve fluorescent protein reporters, where there is a lot of work still to be done. One of his guiding principles is “redder is better”.
I was contacted by a user yesterday looking for a destabilized GFP reporter to monitor promoter repression in addition to promoter activation. This is something I haven’t really looked into much, especially not for use in mammalian cells, so I decided to do some digging. Here’s what I found. Continue reading
This recent paper describes an improved dehydration and clearing method for clearing mouse brains and preserving GFP fluorescence. The use tetrahydrofuran, instead of ethanol, for dehydration, and dibenzyl ether for clearing, and show superior results to the use of BABB or methyl salicylate. They claim this works better than the Miyawaki lab Scale protocol in adult mouse brain.
This is figure 3 from their paper – the left side is conventionally cleared; the right side with their new method:
Reference: Chemical Clearing and Dehydration of GFP Expressing Mouse Brains
Becker K, Jährling N, Saghafi S, Weiler R, Dodt H-U (2012) Chemical Clearing and Dehydration of GFP Expressing Mouse Brains. PLoS ONE 7(3): e33916. doi:10.1371/journal.pone.0033916