[Histonet] Phosphofructokinase histochemical staining in skeletal muscle

[Della Speranza, Vinnie]

I would appreciate hearing from anyone with experience in performing
phosphofructokinase staining in frozen skeletal muscle biopsies.

Is phosphofructokinase especially labile or at risk for false negative
staining?
If so, what circumstances might contribute to this?

Our muscles are transported on ice to the lab and stored in the
refrigerator until they can be embedded and snap frozen.

Our muscles are embedded protruding from gum tragacanth and are snap
frozen in liquid nitrogen cooled 2-methyl butane.

If freezing is delayed, could this contribute to negative staining? How
quickly should these samples be frozen in order to prevent any
diminished enzyme staining?

Other issues to be concerned with when performing this technique?


Sigma Chemical, our regular source, is backordered indefinitely on
Fructose 1.6 diphosphate
Can someone recommend another supplier for this item?

thanks



Vinnie Della Speranza
Manager for Anatomic Pathology Services
165 Ashley Avenue  Suite 309
Charleston, South Carolina 29425
Tel: (843) 792-6353
Fax: (843) 792-8974