[Histonet] Problems with mouse brains

[Ana Resendes]

Hello Kemlo,

I am Portuguese-Spanish....

so we use the word "tampón" that I think I miss translated to
"tamponated"..really do not know if the word exists! but apparently it does
not! :)

I want to mean when you add salt to the formol dilution in destilated water
in order to prepare your 10% buffered formalin...

a believe the word is "buffered"

So sorry..for my wrong translation....I was in a hurry...what I want to say
is THAT YOUR FORMALIN IN NOT WELL BUFFERED
:)
thank you!
and Yes, you should be carefull your brain is well fixated in formalin at
least for 24 hour.

2009/4/5 kemlo <kemlo <@t> f2s.com>

> Agree, poor formalin fixation will allow nuclear bubbling which some
> authorities put down to the effects of heat on the sub optimally stabilised
> proteins. What a marvellous phrase "your formol is not well tamponated".
> Wish I had thought of that......
>
> I will of course steal it if you allow such plagiarism?
>
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu
> [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Ana
> Resendes
> Sent: 04 April 2009 15:16
> To: Randolph-Habecker, Julie
> Cc: histonet <@t> lists.utsouthwestern.edu
> Subject: Re: [Histonet] Problems with mouse brains
>
> have very light H&E staining: first you should check if either your eosin
> or
> haematoxilin are over used
>
>
> nuclear bubbling: can be a lot of things but can be inclusion.
> Brain sections are big so make at least a 24 h inclusion process.
>
> and extensive tissue cracking: could be over heating or bad inclusion. You
> have to check if reagents in inclusors are ok or over passed, I would
> change
> all reagents.
>
> I also have noticed some formalin pigment: your formol is not well
> tamponated
>
>
> 2009/4/3 Randolph-Habecker, Julie <jhabecke <@t> fhcrc.org>
>
> > Folks,
> >
> > I need some input on a problem we're seeing with some mouse brain
> > samples. The samples are from new born mice P0 to P14 which have been
> > fixed in 10% NBF (Fisher brand and well within expiration date) for 72
> > hours. They are then transferred to 70% etoh and processed on an 8 hour
> > process. After the tissue is embedded, we are taking sagital sections,
> > drying them overnight, and then baking overnight at 60C. They are then
> > stained with H&E. This has been working great but now all of a sudden we
> > have very light H&E staining, nuclear bubbling, and extensive tissue
> > cracking. I also have noticed some formalin pigment.
> >
> > My first thought is that there was difference in formalin or time in
> > formalin but that is not the case. I also wondered if the tissue might
> > be exposed to excessive heat. We checked all of the temperatures in our
> > processor, embedding center, and water baths - all were within
> > tolerance.
> >
> > Any ideas what might cause all three artifacts? Could it be from
> > inadequate paraffin infiltration?
> >
> > Any help would be greatly appreciated!
> >
> > Thanks,
> >
> > Julie
> >
> > Julie Randolph-Habecker, Ph.D.
> > Staff Scientist - Director
> > Experimental Histopathology Shared Resource
> > Fred Hutchinson Cancer Research Center
> > 1100 Fairview Ave, N. DE-360 (Please note new location)
> > Seattle WA 98109-1024
> > 206-667-6119
> > jhabecke <@t> fhcrc.org
> >
> > _______________________________________________
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> > Histonet <@t> lists.utsouthwestern.edu
> > http://lists.utsouthwestern.edu/mailman/listinfo/histonet
> >
>
>
>
> --
> Ana Resendes
> DVM, MSc, PhD
> Veterinary pathologist
> Centro de Investigação de Recursos Naturais (CIRN)
> Secção de Anatomia e Taxonomia Zoológicas, Departamento de Biologia,
> Universidade dos Açores.
> Rua da Mãe de Deus, 58 - Apartado 1422
> P - 9501-801 Ponta Delgada (Açores)
> Portugal
> Tel. (+351)  296 650 000 ext. 1109
> Fax  (+351) 296 650 100
> http://www.uac.pt/~pherg/
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
>


-- 
Ana Resendes
DVM, MSc, PhD
Veterinary Pathologist
Centro de Investigação de Recursos Naturais (CIRN)
Secção de Anatomia e Taxonomia Zoológicas, Departamento de Biologia,
Universidade dos Açores.
Rua da Mãe de Deus, 58 - Apartado 1422
P - 9501-801 Ponta Delgada (Açores)
Portugal
Tel. (+351)  296 650 000 ext. 1109
Fax  (+351) 296 650 100
http://www.uac.pt/~pherg/