Which chromogen did you use?

If you work with DAB there is no cause for concern. I for myself would soak the slide at room temperature until the coverslip goes easily off. Then rinse the slide again with xylen until any coverslip-medium is gone. Then coverslip again.

Thanks Gudrun Lang , So, without any re-hydration ?

If the bubbles are only from trapped air in the mountain medium, there is no need for rehydration. Rehydration makes only sense, if you want to rinse the slide again in water or buffer to get rid of eg. too much dye or if you want to destain the slide.

If a slide looks foggy, not clear, it can be due to residual water in the mountain medium. In this case you also go back to absolute ethanol, rinse in two portions of absolute ethanol, then two portions of xylen and then coverslip. So the dehydration is repeated and completed.

prompt reply , thanks Gudrun Lang

Thanks dear Mohamed A.A. Mahdy

Thanks, V.Neubert, I am going to try stirring because it takes at least 3-4 days.

Dear All , I didn't re-hydrated my slides after Xylene treatment and fortunately slides fallen down easily but the problem is that I still have some remnants of the mounting medium on the slides, could you gently tell me how to get rid of this ? Shall I leave my slides uncovered for more 2 days in Xylene or what ? Thanks in advance

To let the slides in xylen and move gently will solve the media. Usually it takes a few hours.

leaving your slides in xylene should not impact on your staining . All you need to do is make sure that the mounting medium you used has been dissolved by the xylene. after that you can go ahead and remount your slides

It is supposed that the xylene will not affect the intensity of the stain. but  you  can leave the slides only over night and the cover slips will fall.

Yes Manal Tawfik Hussein , you are 100 % right , but the problem is that after it fallen down , I have a problem with the remnants of mounting material which is stucking to the slides , that probably will come up with bubbles under the cover slip.

Add to Hasiena Ali suggestion, just remind not good 37c, keep your slides in xylen at lab temp (~25c) for 2-3 days, the coverslips will get off easily.

Thanks a lot Dear Gila Behzaid

Dear V. Neubert, I am using Aquatex mounting material, I left the slides in Xylene for 3 days at 37 C but the slides did not fallen down alone, I used a scalpel blade to remove the slides and it dismounted easily, but the slide was not clear and many  tints still existed behind, on the surface  

Haitham,

If it is really AQUATEX (R)  (as described, e.g. @ http://www.emdmillipore.com/CA/en/product/Aquatex,MDA_CHEM-108562 :  , find further specification and MSDS there...)  you should have mentioned that as an important point in your question.  CITATION of Info-Text:

So I rather guess you would have been successful leaving the slide in warm A. dest (say 37-40°C) and it should have dissolved properly.

Not amused about, Wolfgang

Dear, Wolfgag H.Muss, Thank you so much for your great help, I appreciate it so much, excuse me for not mentioning about that, these are the mistakes the new user always did.

Dear Haitham,

One should not repeat the mistakes. Learning by doing. We all were and sometimes are like this. :)

Generally all chromogens can be mounted in Aquatex. AEC itself is sensitive to xylene and generally organic solvents. There is a permanet AEC option available from some companies, though, meaning organic solvents resistant.

Good luck and read the instruction manuals from time to time

Thanks Dear Ivan, valuable advice to be considered with the long PhD way.

Dear Haitham,

you don't need to apologize (at least to me!) for a you perhaps made only once in your life...

I understand that young(er) and new RG-members -especially if they have no experience in asking questions - may overlook to tell about the most obvious information which might help to solve their problem fast(est). 

RG claims to help "Discover scientific knowledge",  "make it easy for researchers to share and access scientific output, knowledge, and expertise"....but 

it is only possible to get correct answers from colleagues

if at least the most basic information on the problem is given.

After all I nevertheless would like to ask about your real processing schedule (mounting with the Aquatex) - because you told about "rehydrating"....  did you dehdyrate your sections on slides prior to mounting ? which would have been unnecessary (because Aquatex does not require dehdyration of sections when mounting). Have your residues on the slide/section - as you describe - been removed by placing and gently swirling your slide into water (perhaps after getting rid of nasty xylene by swirling in some ethanol.... ?

Regards, 

Wolfgang

Thanks Dear Dr. Wolfgang H. Muss for being still concerned about my experiment.

First, I want to tell you that my slides are of a fish parasite called "Sea louse" . 

Basically, when I said re-hydration, I meant removing the Xylene that has been used in the demounting procedure, as I did at the beginning of my IHC protocol with the Xylene used for dewaxing.

I tried to do so as you explained, swirling my slides in methanol and washing them in DW for 3 and 5 minutes respectively, the result came up with very clear slides with no stain.

V.Neubert,  I meant by no stain that staining were so faint , I used only AEC and  haematoxylin as a counter stain