Hi,
Some report that some mounting media are not compatible with some fluorophores.
For example possibly Vectashield is not compatible with far red fluorophores. Is this correct ? or is this specific possibly to only one type of far red fluorophores eg Alexa Fluor but nor Atto ?
Are there other known compatibility issues between mounting media and fluorophores or FPs ?
Thanks a lot
The compatibility between mounting media and fluorophore is generally due to the anti-photobleaching/antifade reagent used. Each anti-photobleaching reagent works well for some fluorophores, but may quench or not work as well for other fluorophore types.
Any company that makes mounting media should provide a fluorophore compatibility table somewhere in the product literature; for Vectashield, there’s one here (scroll down).
I’m glad you mentioned Vectashield because I was warned of it quenching AF647, but Vectashield’s graphic makes it seem like there’s no issue.
“Effect of Vectashield-induced fluorescence quenching on conventional and super-resolution microscopy” was published April this year. Here’s an excerpt from their discussion section:
We show that one of the most commonly used mounting media for fluorescence microscopy, Vectashield, if used as an imaging buffer, induces quenching of AF647 and its new variant, AF(+)647. As a consequence, the fluorescence emission of both AF647 and AF(+)647 is much less intense in Vectashield (about 15% and 5%, respectively, of the initial values in PBS).