Frozen tissue, could it be possible to make parafin or OCT blocks? - (Jul/10/2014 )

Hi to all!

We have some liver samples in -80 and we need to make some immunostaining.

I am wondering if some of you have you ever tried to make OCT or paraffin samples with frozen tissue?

Defrost and fix?

I can not find any protocol.... :(

Thanks so much!!

-Vemovied-

How were they frozen? If they were frozen in OCT or something similar, I would say you have a chance, but if they were just flash frozen, then thawing will probably cause massive disruption of fine structure, and hence they will be useless for histology

-bob1-

bob1 on Thu Jul 10 21:10:31 2014 said:

How were they frozen? If they were frozen in OCT or something similar, I would say you have a chance, but if they were just flash frozen, then thawing will probably cause massive disruption of fine structure, and hence they will be useless for histology

They are just snap frozen into linq and now there are in -80....

Do you think if I defrost them in PFA, the degradation will be less aggressive?

-Vemovied-

Thaw them as fast as possible to prevent large ice crystal formation. You might want to put them into a glass forming reagent such as DMSO. Perhaps a glutaraldehyde/DMSO mix might be best.

Have a look at the IHC world website. The forums there might be helpful, but they aren't heavily frequented.

-bob1-

Hi:

I have tried this with liver tissue with poor results, the morphology is very poor. I have defrosted the tissue in NBF !0% 24h.

-immunovir-