HELP!
	I am processing small mucosal GI biopsies at a reference path lab
with a quick 24 hour turn around time, so we use a quick schedule and have
had strange vacuoles appear in the mucosal and connective tissue of many
specimens. Originally, the vacuoles were identified in our longer-processed
tissues. These intense cytoplasmic and nuclear vacuole artifacts were found
in glandular epithelium and within the stroma. The three cases were from a
thyroid gland, a uterine biopsy, and a stomach biopsy. These large vacuoles
were located everywhere throughout the thyroid and uterine slides, but only
on one level of the stomach biopsy.  However, now the vacuoles are found
throughout almost all of the GI specimens which are run on a short three
hour cycle.
It isn't apparently related to reagent changes, since the artifact appears
almost all of the time, regadless of how new the reagents are. Furthermore,
the artifact is showing up in almost all mucosal biopsies at sometime or
another. Below is our processing schedule. All tissue is 10 %
NBF-fixed--usually for at least 12 - 20 hours prior to processing.
	If anyone has ideas to alleviate, eradicate, destroy and/or fix this
problem, I'd be grateful. I have included an image on the Histonet.org image
database titled "Vacuole Artifacts" to show you what the problem is. Any
suggestions or orbservations are appreciated.
	Thanks also to Dr. Linda Margraf, Dr. Herb Hagler and the University
of Texas Southwestern Medical Center for the website--what a great idea!
		Jon Kerr


		     
Routine processing steps all done at Vacuum pressure
15 inches Hg 
Station:	Reagent	Time (min)	Temp  	Pmax (psi)
1	10% NBF		35	40* C.		4
2	10% NBF		30	40* C.		3
3	70 % alcohol	30	ambient		1
4	80 % alcohol	30 	ambient		2
5	95 % alcohol	25	ambient		1
6	95 % alcohol	30	ambient		3	
7	100 % alcohol	25	ambient		4
8	100 % alcohol	30	ambient		4
9	Xylene		25	ambient		3
10	Xylene		30	40* C.		2
11	Paraffin		30	56* C.		7
12	Paraffin		45	56* C 		7
13	Paraffin		45	56* C.		7

End of tissue processing, tissue may be taken out of processor


Rapid processing steps for small GI mucosal/ liver/ prostate biopsies.
Station:	Reagent	Time (min)	Temp  	Pmax
3	70 % alcohol	10		ambient	1
4	80 % alcohol	15  		ambient	1
5	95 % alcohol	10		ambient	1
6	95 % alcohol	15		ambient	1	
7	100 % alcohol	12		ambient	1
8	100 % alcohol	12		ambient	1
9	Xylene		15		ambient	0
10	Xylene		20		40* C	3
11	Paraffin		30		56* C	7
12	Paraffin		20		56* C	7
13	Paraffin		20	56* C	7