[Histonet] RE: Histonet Digest, Vol 59, Issue 32 RE PAS DIGESTION

From:"Kay, Karen"

Good Day Tanisha,
We use the following diastase solution and timing protocol.  It is consistent and works very well.

1.	Diastase Solution

	Amylase (Type VI-B from Porcine Pancreas - Sigma A3176)	0.5 g (Stored in fridge at 4C)
	Tap water	50 mL
	Put 50 mL of warm water from the tap (37C) into a coplin jar (check temperature of water with thermometer).  Add 0.5 g of amylase and stir well.  Use once and 	discard.  

            Prewarm this solution in a 37 C degree oven prior to placing slides into it for 1 hour.

Hope this will help you.

Karen J Kay, MLT
Pathology Supervisor
Chinook Health Laboratory
Chinook Regional Hospital
960 - 19 st South
Lethbridge, Alberta, Canada
T1J 1W5
(403) 388 - 6061 (Phone)
(403) 388 - 6067 (Fax)

> Hi All:
> I am in the process of working up a PAS Staining protocol. The PAS 
> stain is working fine, but I am unable to get my tissue to digest. I 
> am using .5g of Diastase in 50ml of DI water. I tried heating the 
> solution in a water bath for 30 min, 1 hour and 1.5 hours and it still 
> looks the same as the one without digestion. Could I be using the 
> wrong Diastase? If so, does anyone have any suggestions for a new 
> product?
> Thanks,
> Tanisha Neely, HT(ASCP)
> AP-Histology/Specimen Management

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