RE: EFFECTS OF DECALCIFICATION OF BONE ON IMMUNO
One thing I forgot to mention...you're working with neoplastic tissue here, and it doesn't always "behave" like normal tissues. Just another dimension to make things interesting...
From: Johnson, Teri
Sent: Wednesday, April 24, 2002 9:25 AM
To: Fessler, Jim
Cc: Histonet (E-mail)
Subject: RE: EFFECTS OF DECALCIFICATION OF BONE ON IMMUNO
There are quite a lot of articles out there in which studies have been done on the effects of different decalcifying agents on immunostaining, in situ hybridization, and DNA/PCR. J Histochem Cytochem 47:703-709, 1999 (Alers, et al) discuss the latter two. Their literature cited may offer a potential list for you to obtain additional references. Alternately, another article J Histochem Cytochem 50:113-115, 2002 (Brown, et al) showed they were able to show that 5% formic acid treatment preserved DNA in metastatic prostate cancer specimens. I have an article on Zebrafish where they tested various decalcifiers and found that 0.5M EDTA maintained the best immunoreactivity (BioTechniques 32:296-298, Feb 2002).
My advice would be to obtain some control tissue (we always used tonsil) and after fixation, subject it to your decal fluid for your routine time. Wash and process in tandem with a usually processed tonsil. Re-run your MIB-1 using both pieces of tissue and see if there is any difference in staining. It might be even better to have them on the same slide so you're sure they receive the same treatment under the same conditions.
I think the bottom line (for researchers anyway) is that formic acid decalcifying provides "good enough" results with maintaining immunoreactivity, and is quicker than EDTA.
Gayle Callis and Donna Montague are good contacts (probably among many others) for additional information.
Managing Director Histology Core Facility
Stowers Institute for Medical Research
1000 E. 50th St.
Kansas City, Missouri 64110
From: Fessler, Jim [mailto:firstname.lastname@example.org]
Sent: Wednesday, April 24, 2002 7:50 AM
Subject: EFFECTS OF DECALCIFICATION OF BONE ON IMMUNO
Good Morning all, happy spring....
Anyone know of any articles published concerning the effects of
decalcification on antigenicity of tissue used for immuno?
I am running Ki67 (mib-1) on osteosarcomas and chondromas and there seems to
be a loss of positive staining on most, but not all of them. I suspect that
the decalcification process has effected them.
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