[Histonet] R: EDTA decalcification tissue issues

[MANTERO Stefano RIC]

Good Morning Kelly,

I work in a research institute that works with murine samples; In our laboratory we have adopted two descaling protocols: one with ion exchange resins/acid solution while the other uses EDTA.
The EDTA descaling system is slower but keeps the nucleic acids much better.
We use an 14% solution of tetrasodium EDTA in water buffered to pH 7.4-7.6 with acetic acid, the solution is left for several days (from 5 to 10 depending on the density and size of the bone) with daily solution changes.

Have a good work

Stefano

Dott. Stefano Mantero

Human Genome Laboratory  CNR-IRGB
c/o Humanitas Research Hospital

Via Rita Levi Montalcini (Ex Via Dainese)
20090 Pieve Emanuele (MI)

Ph. +39 02 8224 5164 (desk)
Ph. +39 02 8224 5177 (lab)
Fax +39 02 8224 5191



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-----Messaggio originale-----
Da: Pairan, Kelly via Histonet [mailto:histonet at lists.utsouthwestern.edu]
Inviato: venerdì 10 novembre 2017 12:39
A: histonet at lists.utsouthwestern.edu
Oggetto: [Histonet] EDTA decalcification tissue issues

Good Morning,

Recently, our lab has been working on validating an EDTA method of decalcification.  When we ran the IHC's on the decalcified bone block, the majority of the tissue lifted off the slide.  We use the Leica Bonds for our IHC staining.  Does anyone else have a hard time getting EDTA decalcified tissue to stay on positively charged slides during IHC runs?  Do you have a trick you use?  We really did not have this big of a problem when we were decalcifying with Formical or Nitrical (big bones only).


Thanks for your help,

?Kelly Pairan, HT(ASCP), QIHC(ASCP)
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