Histonetters,
	A couple months ago I wrote asking how to do the Elastin 
stain.  I received instructions (thank you very much!) and now have 
experience with it .  BUT I am having a big problem with it -- lack 
of consistency.  Slides cut at the same time, from the same animal, 
same tissue (lung) have been used to work out the conditions in our 
lab.  One time it will work beautifully, the next time using the same 
times -- there is NO staining.  None. I just don't get it.  I have 
included the procedure and solutions below.
	Can any one give me some insight as to why this is happening? 
FYI... lung tissue was immersion fixed in 4% paraformaldehyde prior 
to paraffin embedding which was done by our histo lab.  I thank you 
in advance for your help.

Kathie


Verhoeff Stain to visualize Elastin

… Rinse in Distilled Water. Stain in Verhoeff's working solution for 30
minutes.
… Rinse in tap water until clear.
… Differentiate if 2% Ferric Chloride from 2-8 minutes (monitor it under
scope) - we used 5 min
… Rinse in Tap water until clear.
… Then dehydrate, clear & mount.

Solutions for the Verhoeff portion

Verhoeff Stock       	20g Hematoxylin (CI-75290) / 400 mL 100% ETOH
(filter after dissolved)

10% Ferric Chloride  	26g Ferric Chloride / 260 mL DH20

Verhoeff Working Stain	330 mL Verhoeff Stock / 130 mL 10% Ferric chloride /
130 mL Lugols Iodine

2% Ferric Chloride	120 mL 10% Ferric Chloride / 480 mL DH20