Is there a way to rescue my tissue sections after they have dried out during primary antibody incubation overnight? If I boil the slides in antigen retrieval solution and repeat the staining procedure will I still have spurious staining?
So, I read all the above suggestions.....my sections had dried out during blocking at room temp....i still went ahead and did the full protocol and the staining worked beautifully. so i would say its worth finishing the staining.
I would try to rehydrate in 0.1 PBS for several hours, sometimes it works. At least you should be able to recover your tissue.
cheers, refik
To Amanda Photenhauer:
I am wondering what's the term "rescue" means? If you means connection to secondary antibody and following coloring, the answer may be "Yes". You could rehydrate them as Refik Kanjhan said.
But the most important outcome of IHC is the conditions of each process. If the slices were dried out, the problem is the concentration of primary antibody is inconsistent, and there are no stripping protocol as I know; Therefore, it become non-reproducible even there are good results (maybe spurious results as you said). In my opinion, do it again directly is much more efficient way.
Yes, those slides will most likely be ruined I'm afraid. A good way of preventing this is to cover the slides with a strip of parafilm after you put your antibody solution on. That way you only need to use at most 150 ul for an entire slide, and, provided you keep them humidified, they will definitely not dry out.
When biological samples dry, they form covalent bonds, both within the sample and with the glass slide, that are very hard to break. As such, unless the sections are extremely precious, I would just make new sections off the block and start over. Even with some aggressive rehydration, the results will be very hard to interpret, and in no way publishable.
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