trephine bone marrow biopsy in EDTA

Zbigniew Rudzki mprudzki at cyf-kr.edu.pl
Tue Mar 9 10:07:55 EST 1999

(the question below is more sci.med.pathology-like, but since people in
b.m.m-r have frequently astonishingly interdisciplinary background I dare to
post it here as well):

I decalcify bone marrow trephine biopsies in 6% (w/v) EDTA at 37 C.  EDTA is
dissolved in ddH20, without pH adjustment.  Six % seems to be saturating
solution.  The trephines are fixed in 10% buffered formalin prior to
decalcification. The morphology of paraffin-embedded tissue is nice and the
procedure allows to perform Leder's chloroacetate esterase (the enzymatic
activity is lost if any acid is involved in tissue processing).  Also the
antigens are generally nicely preserved.

I have one problem I can not overcome:  the time.  It usually takes over 5
days to get tissue sufficiently soft to cut it.  This seems unacceptable and
frustrating, both for hematopathology lab and clinicians.  I strongly
suspect that the procedure is flawed by some trivial technical error.

Could anyone offer me some suggestions ?
(no acids !!!)

Zbigniew Rudzki, MD
Dept. Pathol. Jagiellonian Univ., PL

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