I mounted DAB stained sections onto a slide and used DPX as a mountant. There seems to be some sort of a problem in some of these sections post mounting. I would like to remove DPX and re-mount the sections. Has anyone done this previously? Is there a standardized protocol for this? A few important points :-
1. The sections were not dehydrated with ethanol dilutions prior to mounting.
2. The slides are not old. They were mounted less than a month back.
3. The immuno has worked in most of the sections/slides. Its only some slides that are problematic.
Thanks
Sourish
In general to re-coverslip a slide you have to soak the slide in the solvent of the mounting media. So for DPX you have to soak the slide in xylene or xylene-substitute until the coverslip can easiliy be removed. This may take 1-3 days, depending on the age of the slide. Then rinse the slide well in xylene until the mounting media is all gone. If you doubt about proper dehydration, rinse in 2-3 portions of abs. ethanol, then 1-2 portions of xylene and coverslip again.
If mounting media is still on the slide and you go into diluted ethanol a white haze will be seen.
In general to re-coverslip a slide you have to soak the slide in the solvent of the mounting media. So for DPX you have to soak the slide in xylene or xylene-substitute until the coverslip can easiliy be removed. This may take 1-3 days, depending on the age of the slide. Then rinse the slide well in xylene until the mounting media is all gone. If you doubt about proper dehydration, rinse in 2-3 portions of abs. ethanol, then 1-2 portions of xylene and coverslip again.
If mounting media is still on the slide and you go into diluted ethanol a white haze will be seen.
Hello, put the slides you want to remount in xylene for 10 minutes. Remember the container will be made of glass. Then follow the method as described by Gudrun Lang above.
Hi There . The easiest way to do it is to allow the slides to remain in a coplin jar or staining dish with Xylene . Allow it to stand overnight or for however many days it takes for the coverslips to come off. Thereafter follow the procedure as outlined by Gudrun Lang
In general, you have to put slides in xylene for 2-3 hrs with shaking or overnight without shaking.
Hi just dip the slide in the xylene for 5- 10 minutes.This procedure we are routinely doing in our laboratory.
The best solving material for the slide glues is Xylol 100%, Put the slides in it until the DPX glue and coverslips will be detached
To salvage the DPX mounted sections it is safer to leave slides in vertical coplin jar with pure xylene till the cover slip gets detached. Once the DPX dissoves the coverslip slips to bottom of the jar on its own weight, without causing any damage to the section.
break the coverslip without disturbing the section preferably with a wide bore needle. then drop in xylene. This will work in an hour for freshly mounted sections.
Yes, I agree with Gudrun Lang. But take care to see that the section is not washed off the slide.
Soaking the slide in pure xylene till the coverslip slips down would be the best method to recover the section.
Yes I agree xylene is the best. you can take xylene in coplin jars and dip the slide in it. and check till the coverslip has slipped off and left behind in the coplin jar. have patience and don't try to remove it forcibly. just remove the slide by using forceps and check if the coverslip is still there.
place them in xylene (glass coplin jar not a plastic one) , cover slip will detach itself and after that you can remove your slide and remount it. If the problem persist change mountant , get a new bottle of DPX.
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