[Histonet] Re: Bird head stored in 70% alcohol and possible decalcification

[Rui TAHARA]

Thank you for all the helpful suggestions about this topic. 
My sample has been in the fixative until it would be decalcified. 

Thank you again,

rui  

> From: ree3 <@t> leicester.ac.uk
> To: ruio7 <@t> hotmail.com
> Subject: RE: [Histonet] Re: Bird head stored in 70% alcohol and possible	decalcification
> Date: Tue, 10 Mar 2015 09:49:00 +0000
> 
> Leave  it  in   formalin for  as  long  as possible..........................good  luck
> 
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Rui TAHARA
> Sent: 04 March 2015 23:39
> To: gayle.callis <@t> bresnan.net; histonet <@t> lists.utsouthwestern.edu
> Subject: RE: [Histonet] Re: Bird head stored in 70% alcohol and possible decalcification
> 
> Thank you for helpful suggestions. 
> I have further questions. 
> Yes, I have a bird head (probably 1 cm X 1 cm ) stored in 70 % ethanol.
> But i have a similar size bird head fixed in 3.7% formalin for over night and am actually processing the head to store in 70% ethanol since my lab is just ordering the decalcifying solution. I need to decalcify this sample later. 
> But i am wondering if it is better to keep the sample in formalin for a week or so till i get the decalcification solution or i should store it in 70 % ethanol and then fix it for a few days again later? 
> I am afraid that longer fixative time would affect the sample somehow (e.g. the sample become too rigid?) 
> 
> Thank you, 
> 
> rui 
> 
> > From: gayle.callis <@t> bresnan.net
> > To: histonet <@t> lists.utsouthwestern.edu
> > Date: Wed, 4 Mar 2015 16:25:28 -0700
> > Subject: [Histonet] Re: Bird head stored in 70% alcohol and possible	decalcification
> > 
> > You wrote: 
> > 
> >  
> > 
> > I have an adult bird skull that fixed with formalin and then has been 
> > stored in 70% ethanol.
> > 
> > I have seen the post that the sample stored in 70% ethanol can be 
> > walking back through to series of ethanol to water and can be 
> > decalcified if it needs to be.
> > 
> >  
> > 
> > I am wondering if anybody has done this and there is any side effects 
> > from decalcification after going through dehydration and rehydration 
> > of a sample compared to a general straight forward protocol from 
> > decalcification to dehydration?
> > 
> > **********************************************************************
> > ******
> > **********************************************************************
> > ******
> > ********************************
> > 
> >  
> > 
> > I have, in the past, when a weekend arrive, I interrupted acid bone 
> > decalcification by removing it from acid decalcifier, a quick water 
> > rinse and immersed into 70% alcohol before returning bone to fresh 
> > acid decalcifier the next working day. The bones always decalcified 
> > without problems but I am sure the decalcification took longer since 
> > partially decalcified bone had to rehydrate.  I later learned more 
> > about dipolar (hope I said that correctly) alcohol slowing and/or stopping ionization of calcium
> > and ceased using 70% alcohol to interrupt acid decalcification.   I now use
> > NBF to interrupt decalcification.  Interestingly, I learned the alcohol
> > technique from the AFIP bone pathology lab.     
> > 
> >  
> > 
> > Alcohol is put into Perenyi's  nitric acid decalcifying solutions to slow
> > down or control very rapid nitric acid decalcification.     
> > 
> >  
> > 
> > You did not say how big the bird skull was?   I suggest immersing the skull
> > back into NBF to let it totally rehydrate for several days (depending 
> > on skull size and if the brain is present).  I suggest changing NBF if you
> > rehydrate longer than a day.   You don't need to go back through an alcohol
> > gradient since many processing schedules have tissue samples  going from NBF
> > directly into 70%.   If you leave residual alcohol in the bones, the acid
> > decalcification could be slower and hopefully not retarded in any way.
> > 
> > 
> >  
> > 
> > It certainly is worth a try.   Good luck.
> > 
> >  
> > 
> > Gayle M. Callis
> > 
> > HTL/HT/MT(ASCP)
> > 
> >  
> > 
> >        
> > 
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