[Histonet] rat myelofibrosis questions

[Gayle Callis]

Missy,

Concentration of EDTA will depend on which EDTA you use.

EDTA, disodium or EDTA without any sodiums attached only goes into solution 
at around 10% or 10g/100 ml, pH 7.0 to 7.6 and people often have to use 
sodium hydroxide to make it go into solution which should also adjust the 
pH up to 7.0.

If you use EDTA tetrasodium you can get the concentration very high at 14% 
or more, but you must adjust the pH down.  We use glacial acetic acid for 
this purpose (bone expert Webb Jee used this EDTA decalcification method 
for bone).  EDTA tetrasodium dissolved in water or buffer has a pH of 
approximately 9 to 11, so the pH should be adjusted down to pH 7.4 or 
so.  If the pH remains very alkaline, then the alkaline sensitive protein 
linkages can be damaged.

You may want to look at the femur too, since you could use the 
contralateral side (femur) for your other test.   Have you done a 
literature search on mouse ilium just to see what is out there.  Google 
Scholar is a good place to search if PUBMED becomes cumbersome. Sternum is 
another bone marrow biopsy site in humans - just a thought here.

We used to do fibrin staining on acid decalcified bone marrow biopsies 
without problems rather than use the slower EDTA decalcification.



If you useAt 09:37 AM 11/2/2006, you wrote:
>Hi all,
>
>Sorry to bother again, I just have a quick question.  I am looking for
>myelofibrosis in rat, we had intended to look at the femur,  however we got
>to talking and brainstorming and were wondering if the ilium would be a
>better place to look.  Since the iliac crest is the location of bone marrow
>biopsy in human, and that way we can "double dip" on the animals and use the
>femur for another assay.  Do you think it will work or has any one gotten
>positive results using the ilium?
>Also, regardless of wehter we use femur or ilium, which concentration of
>EDTA is optimum for decal?
>
>Missy
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Gayle Callis
MT,HT,HTL(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367
406 994-4303 (FAX)