I have to do post-section staining of my slides. With the tissue freezing medium present, my specimens were washed away during staining. So, how do I dissolve this medium without affecting my sectioned specimen? How do I fix the specimen onto the slide?
Normally, I incubate the fresh section for 30 sec. with 100% Ethanol. Should not influence the epitope availability and keep the tissue on the slide!
Hello Rathnnavel.
As in the discussion suggested above, you should use coated glass slides like Superfrost Plus, or others. OCT is soluble in water, so you have to wash the sections in PBS or similar to get rid of it. We use Histobond slides from Marienfeld.
Depending on the thickness of your slides you could also perform free-floating stainings in wellplates and coverslip afterwards . This is good for slices thicker than 30um, which are not so fragile.
Greetings
Rathnavel, the responses you have gotten above are very good, informative and practical. I addition, your technique for drying and washing needs fine tuning, so that you use the most gentle method/agitation once you have fixed the tissue to the slide. if you are placing your slides in a glass jar and agitation with a shake, sometime your tissue will detach when pick out the slides (e.g. holding basket). Please update when you succeed in reducing the frequency of detachments....
Thank you everyone. This thread was really helpful.
I didn't use a Superfrost slide and will use one from now on. I think that will help keep the tissue attached.
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