RE: [Histonet] Re: LacZ Staining of Tumors

From:"Johnson, Teri"

Markus, we have successfully stained with x-gal, then decalcified with formic acid (ImmunoCal) and then paraffin processed. In our hands, if we did the x-gal technique first and then decalcified with EDTA, the blue positive staining came out. It does not using formic acid.

I have seen published reports successfully x-gal staining by decalcifying with EDTA first, then staining with x-gal and paraffin processing.

Beware doing this technique on bone though. Osteoclasts in normal bone will stain positively with the X-gal staining procedure (ref. Histol Histopathol (2007) 22: 971-976 B-Galactosidase staining on bone marrow. The osteoclast pitfall)

Best wishes,

-----Original Message-----
From: Markus F. Meyenhofer []
Sent: Thursday, October 16, 2008 2:35 PM
To: Johnson, Teri
Subject: Re: [Histonet] Re: LacZ Staining of Tumors

Thank you for this note.
I want to do this on bone (Paraffin).
Do you think whole mounts on bone will work, plus de-cal? Regards, Markus
----- Original Message -----
From: "Johnson, Teri" 
Sent: Thursday, October 16, 2008 2:02 PM
Subject: [Histonet] Re: LacZ Staining of Tumors

Melissa wrote:

>>I am looking for a protocol to fix and stain in paraffin embedded
>>subcutaneously grown mouse tumors for LacZ.  Can anyone help?  M.<<

Far as I know, one cannot stain paraffin embedded material with x-gal. We will do either whole mount X-gal staining, post-fix, and then paraffin process and section, or we will fix, sucrose cryoprotect, freeze and then cryosection to do the X-gal staining. Send me an email if you want to have our protocols for doing either.

Teri Johnson, HT(ASCP)QIHC
Managing Director Histology Facility
Stowers Institute for Medical Research
1000 E. 50th St.
Kansas City, MO 64110

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