Jump to content

  • Log in with Facebook Log in with Twitter Log in with Windows Live Log In with Google      Sign In   
  • Create Account

Submit your paper to J Biol Methods today!
Photo

Frozen tissue, could it be possible to make parafin or OCT blocks?

frozen oct paraffin ihc

  • Please log in to reply
4 replies to this topic

#1 Vemovied

Vemovied

    member

  • Active Members
  • Pip
  • 14 posts
0
Neutral

Posted 10 July 2014 - 01:53 AM

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!!


#2 bob1

bob1

    Thelymitra pulchella

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 5,728 posts
399
Excellent

Posted 10 July 2014 - 01:10 PM

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



#3 Vemovied

Vemovied

    member

  • Active Members
  • Pip
  • 14 posts
0
Neutral

Posted 11 July 2014 - 12:32 AM

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?


#4 bob1

bob1

    Thelymitra pulchella

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 5,728 posts
399
Excellent

Posted 11 July 2014 - 01:20 AM

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.



#5 immunovir

immunovir

    member

  • Members
  • Pip
  • 2 posts
0
Neutral

Posted 01 October 2014 - 03:30 AM

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.






Home - About - Terms of Service - Privacy - Contact Us

©1999-2013 Protocol Online, All rights reserved.