Fluorescence Stained Sections - Later Examination Possible? (Nov/02/2007 )
hallo,
Is it possible to store fluorescence stained sections and evaluate them at a later date, or should they have to be evaluated immediately after staining? when its possible to store them, which conditions should be chosen?
thanks to all
Are these fixed? Usually it should be. Do you have a good seal to prevent evaporation?
Is it possible to store fluorescence stained sections and evaluate them at a later date, or should they have to be evaluated immediately after staining? when its possible to store them, which conditions should be chosen?
thanks to all
later is possible if sections are fixed; against bleaching use DABCO; however, quality of fluorescence decreases over the time
hallo,
thanks for replies. my stained sections are fixed in DAKO Fluorescent Mounting Medium.
fixation isnt really the problem (if you didnt fix them at some point you'd get dodgy data) the thing you really have to look out for is the photobleaching - wrap the slide trays in tin foil and keep them covered at all times (we also store ours in the cold room) - you should be able to get a week of decent pictures (depending on the strengh of your staining / antibody) after that you will see a sharp decline (i never use slides older than two weeks as you also get bleeding of the fluorochrome and thus fuzzy images.
dom
Hi,
from my experience - after drying them properly you could store them at-80C for a very long time. We got a nice (paper quolity) picture after even store at -20 for 6 month. But don't forget to seal them with a nail polish.
from my experience - after drying them properly you could store them at-80C for a very long time. We got a nice (paper quolity) picture after even store at -20 for 6 month. But don't forget to seal them with a nail polish.
this is a new one to me
how does the vectasheild react to freezing?
by drying do you mean removal of the pbs?
does the tissue crack?
dom
Dominic,
so far it worked. By drying I mean let them stay for a while (in the dark) after coverslipping before applying the nail polish,so the cover glass not slipping.
Right now I work with prolong antifade reagent from Invirogen (with or without DAPI), but used a vectashield before too....
hallo,
i normally use fluorescent mounting medium from dako. does some of you have same experience with that? long-time storage...
Better to analyze immunofluorescent sections immediately as fluorescence fades over time even with the anti-bleach mounting media.
I have stored them in 4C and usually analyzed them in a week but sometimes within a month and they are fine.