My photophysics reading for this week has been "Quantitative FRET Analysis With the E
In this article, some scientists from various institutions within the Scuola Normale Superiore in Pisa propose a new fluorescent protein pair (E0GFP-mCherry) which they argue is particularly useful for FRET experiments in living cells.
The link here to Kristen's paper should be obvious and, in case you're interested, I got a bit carried away trying to find a link to Michael's. I might have come up with a link that actually has some conceptual substance by Tuesday, but in the meantime here's my best effort: mCherry is a monomeric derivative of DsRed, a red fluorescing protein derived from stony coral. Michael's paper features HcRed, which is also red fluorescing and also derived from an anthozoa, but is dimeric & found in nature in an anemone, not a coral.
back on track... my article describes the quantitative determination of FRET efficiency for this pair using fluorescence lifetime imaging spectroscopy (FLIM) & acceptor photobleaching (APB). I found the format a bit "biology-esque" (with descriptions of method focussing on technique used for, say, cloning proteins) but these were all new ideas to me so it was interesting and I coped! I recommend skipping to the treatment of FLIM & APB in the Results section which was somehow more conceptual.
The authors also talk about "the recent phasor approach" to FLIM, which I had to go here to understand.
