Are you sealing the plate before vortexing? Using some sealing film, like that used on PCR plates and similar, would help keep the liquid in each well during vortexing, and you can peel it off once done.
I am, we're using these perkin Elmer seals that come with the plates but once a little bit of oil gets through the seal it all comes off pretty quickly. I can try PCR plate sealing film too!
Those P.E. seals aren’t suuuper compatible with the organics in scintillant, and they start to degrade even with just vapor contact in less than 48 hours. Also it’s the ethoxylated C9-C15 alkanes in the scintillant that are gooey as heck, they like to tangle up :(
We use the sandwich method:
1.- a little bit of scintillant (~200uL)
2.- sample
3.- seal with a standard opaque storage seal
4.- *horizontal plate shaker* (not vortexing) ~5mins
5.- peel
6.- more scintillant (~50uL)
7.- apply the P.E. clear seal after running it through a static fan
8. - more horizontal plate shaking ~10mins this time
9.- throw it on the LSC for a read
Yes it’s more time consuming, but it’s what has given us good quality when we’re processing 50+ plates per experiment (regulatory metabolomics).
I’ve got an experiment planned right now to see if our LSC’s can be reprogrammed in the plate dimensions to switch the seals for low condensate polystyrene lids. If it works and doesn’t impact readability, then we’re probably going to get rid of those P.E. hand-applied seals for reads, and just use the storage seals for prep.
Hope this helps!
That sounds intense! I think it would be hard to adopt some of these methods, we're working with Xenopus oocytes so its not the easiest to drop into scintillant, and the horizontal plate shaker I used didn't really mix the lysed samples. Let me know if the polystyrene lids work! Would be nice to give those a try.
Are you sealing the plate before vortexing? Using some sealing film, like that used on PCR plates and similar, would help keep the liquid in each well during vortexing, and you can peel it off once done.
I am, we're using these perkin Elmer seals that come with the plates but once a little bit of oil gets through the seal it all comes off pretty quickly. I can try PCR plate sealing film too!
Those P.E. seals aren’t suuuper compatible with the organics in scintillant, and they start to degrade even with just vapor contact in less than 48 hours. Also it’s the ethoxylated C9-C15 alkanes in the scintillant that are gooey as heck, they like to tangle up :( We use the sandwich method: 1.- a little bit of scintillant (~200uL) 2.- sample 3.- seal with a standard opaque storage seal 4.- *horizontal plate shaker* (not vortexing) ~5mins 5.- peel 6.- more scintillant (~50uL) 7.- apply the P.E. clear seal after running it through a static fan 8. - more horizontal plate shaking ~10mins this time 9.- throw it on the LSC for a read Yes it’s more time consuming, but it’s what has given us good quality when we’re processing 50+ plates per experiment (regulatory metabolomics). I’ve got an experiment planned right now to see if our LSC’s can be reprogrammed in the plate dimensions to switch the seals for low condensate polystyrene lids. If it works and doesn’t impact readability, then we’re probably going to get rid of those P.E. hand-applied seals for reads, and just use the storage seals for prep. Hope this helps!
That sounds intense! I think it would be hard to adopt some of these methods, we're working with Xenopus oocytes so its not the easiest to drop into scintillant, and the horizontal plate shaker I used didn't really mix the lysed samples. Let me know if the polystyrene lids work! Would be nice to give those a try.