Hi,
Is anyone using mNeongreen? We have recently switched to making our translation fusions with it. Though we haven’t compared the same protein, so I would hesitate to comment on brightness, it seems that mNeongreen bleaches much quicker than GFP.
Has anyone else noticed this, or have any other experience with mNeongreen?
Vic
I’ve done a bit with mNG, but haven’t done any direct comparisons. Not sure if you came across this BioRxiv paper, but the Goldstein lab did a really nice comparison of a range of fluorescent proteins: http://www.biorxiv.org/content/early/2016/02/19/040279
Take home messages were: i) mNG was not as bright as expected; ii) using 488 nm illumination, GFP and mNG performed similarly (brightness, photobleaching, etc.); iii) there seems to be gene-to-gene variability: for some fusions GFP was brighter, for other fusions the mNG was brighter; and iv) lower background autofluorescence is observed at 514 nm illumination, so for lowly expressed genes, they recommend mNG with this illumination. It’s the best combo of brightness, signal-to-noise, and photostability.
Hi Vic,
I was going to point you to our preprint, but Jordan beat me to it. The only thing I would add that’s not in the paper is that anecdotally, mNeonGreen seems to be more prone to germline silencing than GFP, so that’s something to be aware of if you’re working on the germline or early embryo.