[Histonet] Bone!

Tue May 26 06:19:00 CDT 2009

As I say I don't have experience of preservatives but question if the
effects of ethanol (70%) over a long time may also 'overfix' the bone?
Anyway 70% ethanol has perfectly preserved me!!

Kemlo Rogerson	
e-mail kemlorogerson <@t> nhs.net if not at work.			
DD   01934 647057 or extension 3311     Mob 07749 754194; 
Embrace uncertainty. Hard problems rarely have easy solutions. --Jonah
Lehrer
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-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Rittman,
Barry R
Sent: 22 May 2009 13:21
To: histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] Bone!

Lets face it there is no really ideal medium for long term storage.
However this depends on what techniques you want to carry out later and
when.
I agree that 10% NBF is a good fixative for bone, I would definitely not
advise leaving it in this fixative for extended periods of time.
The reason is that formalin continues cross linking.
This means that many stains and immunohistochemistry may no longer be
possible depending on the amount of time left in fixative.
Also Kemlo is correct in that temporary formalin bonds can be broken by
extended washing in watre. This only applies to the weak temporary bonds
and not to the more stable bonds. The longer the fixing the greater the
number of stable bonds.
I would recommend having a mixture of 70% ethanol plus glycerin up to
10-20%.  The glycerin will prevent any drying out of the solution over
extended periods of tim.
Barry

________________________________________
From: histonet-bounces <@t> lists.utsouthwestern.edu
[histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Kemlo Rogerson
[Kemlo.Rogerson <@t> waht.swest.nhs.uk]
Sent: Friday, May 22, 2009 2:24 AM
To: Robert Edward Pogue
Cc: histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] Bone!

Probably of all the fixatives 10% neutral buffered formalin is least
likely to overfix and I've left tissues in it for years. Formalin, as
you know, is one of those fixatives that you can wash out. There are
preservatives, rather than fixatives, you could try but I don't have
experience of those; my best advise is formalin but make sure that it is
neutral and buffered. I think the effects of pH would be more damaging
than the relatively 'soft' formalin fixation.

Kemlo Rogerson
e-mail kemlorogerson <@t> nhs.net if not at work.
DD   01934 647057 or extension 3311     Mob 07749 754194;
Embrace uncertainty. Hard problems rarely have easy solutions. --Jonah
Lehrer This e-mail is confidential and privileged. If you are not the
intended recipient please accept my apologies; please do not disclose,
copy or distribute information in this e-mail or take any action in
reliance on its contents: to do so is strictly prohibited and may be
unlawful.
Please inform me that this message has gone astray before deleting it.
Thank you for your co-operation

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Robert
Edward Pogue
Sent: 21 May 2009 16:33
To: Kemlo Rogerson
Cc: histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] Bone!

Thanks Kemlo...

I thought of that, but was worried about over-fixation,- what do you
think?

Redward.

------------------------------

In 10% neutral buffered formalin?

Kemlo Rogerson
e-mail kemlorogerson <@t> nhs.net if not at work.
DD   01934 647057 or extension 3311     Mob 07749 754194;
Embrace uncertainty. Hard problems rarely have easy solutions. --Jonah
Lehrer This e-mail is confidential and privileged. If you are not the
intended recipient please accept my apologies; please do not disclose,
copy or distribute information in this e-mail or take any action in
reliance on its contents: to do so is strictly prohibited and may be
unlawful.
Please inform me that this message has gone astray before deleting it.
Thank you for your co-operation

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Robert
Edward Pogue
Sent: 21 May 2009 14:47
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Bone!

Friends,

Can someone recommend to me how to store bones (juvenile rat femur) that
I have fixed and decalcified, but am not yet ready to cut (they're for
vibratome, so not embedded).

Thanks!

Redward.

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