the dilute concentration for DAPI staining - (Feb/05/2007 )
This is the website (www.vectorlabs.com). Search for VECTASHIELD HardSet Mounting Medium with DAPI .
I'll be honest i've never used the hardset mediums - i tend to stick with liquid under the coverslip and a rather fetching shade of red nail varnish around it - but each to their own
dom
dom
thanks for keeping attention . could you tell what kind of liquid you stick with, here I didnt understand well what means stick either. Because I worry if that is just normal buffer liquid or at least the antifade mounting liquid? Techinically in our lab we also do like you and seal the margin of coverslips with nail varnish.
And if I must buy the readymade mounting media from VECTOR? In our lab they traditionally confect the mounting media themself with Glycerin, PBS and DABCO.
cherry
When i said i stick with i meant it in the way that it is my preferred choice and what i almost always use rather that a form of glue i use on the cover slip.
when you use the vectorshield you just dab off any excess pbs left from your experiment (i tend to tap the slide on a piece of tissue on the desk) put 10 ul vectorshield on the slide next to the tissue (dont want to scratch off your experiment at the last minute) then drop the coverslip on using curved tweezers as a guide. let it settle for ten minutes removing any bubbles with the tweezers (in darkness - photo bleaching) then paint around it with the nail varnish.
hey presto - problem solved
dont worry about asking lots of questions - its the only way to learn (you should see the way i pester people when i dont know something.)
dom
thanks for patience, it is so clear about the procedure.
And if I must buy the readymade mounting media from VECTOR? In our lab they traditionally confect the mounting media themself with Glycerin, PBS and DABCO.
cherry
well you could make it yourself but that takes you back to square one as you'll need the concentration (if you must do it this way a simple assay 1/50 1/250 1/500 1/1000 1/5000 on your favorite tissue type would give you the answer).
personally tho id just take a sharp breath and buy the stuff
dom
I have used the Hardset from vector labs and found it to be good. In our old lab, we used to make up mounting media and used to counter stain with DAPI separately. Somehow due to the ease, I prefer the Vectashield.
Ofcourse, you do need to go with the lab's preference.
Good Luck !!!
thanks a lot to you both for your attention and sharing experience. I will try.