Re: Trap stain for Osteoclasts

<< Previous Message | Next Message >> (Hermina Bogerink), histonet <>
Content-Type:text/plain; charset=us-ascii


Here is a reference for the Trap method using regular paraffin:

  Erlebacher and Derynck  J.Cell Biol 132:195-210, 1996

The protocol says to incubate for one hour at 37 degrees Celcius,
however I have found that it is necessary to incubate for at least three
to four hours.
The solution consists of:

For 50.0 ml:
0.2M sodium acetate = 0.82 gm
50mM L(+) tartaric acid = 0.58 gm
pH 5.0
Incubate secations for 20 minutes at room tempertature, then add:
0.5mg/ml naphtol AS-MX phosphate = 25mg
1.1mg/ml fast red TR salt = 55 mg
Incubate for three to four hours at 37 degrees C.

After deparaffinizing your slides, incubate them in the above solution.
The osteoclasts will be a bright red.  For contrast, I counterstain with
Mayer's hematoxylin that I make up myself.

Hermina Borgerink, BA, HT(ASCP)IHQ
Wake Forest University Baptist Medical Center
Medical Center Blvd
Winston-Salem, NC 27157
ph 336 716-1538

<< Previous Message | Next Message >>