Re: Diastase/Amylase digestion

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From:rkline@emindustries.com (by way of histonet)
To:histonet <histonet@magicnet.net>
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Kerry,

The fact that you are not getting good results with other manufacturers
diastase, is giving me the red light that something else is going on such
as fixation, tissue processing,and/or buffer.

I'm an ex-spitter and have no experience with other digestion.  And I guess
at this point with all the pathogenes and AIDS other methods for digestion
are probably more appropriate.

I did a little research.  According to Lee Luna's book,  the reason why
spit works well is because for some reason amylase dissolves glycogen
better.  Amylase is commercially available.  You should be looking for an
amylase from pancreas.  I checked BDH's catalog and found that the amylase
they have is from bacteria.  You still may want to give them a call and see
if they have an amylase from pancreas.  It may not listed in the catalog.

If you would like a copy of the procedure for making the digestion solution
from amylase let me know.  I'll be happy to fax it to you.

Rande Kline HT (ASCP)
Technical Services
EM Science








bbracing <bbracing@silk.net> on 01/25/99 01:37:00 AM

To:   Histonet <histonet@pathology.swmed.edu>
cc:
Subject:  Diastase/Amylase digestion




A little help is needed.
We used to get a very good diastase from BDH (diastase of malt) that worked
as good as, or better than "spitting" on the slides for glycogen digestion.
The last several bottles have not worked well.  We have also tried material
obtained from Fisher and Sigma  again with very poor results. We used to
use a 0.25% in sailine for 10 min.at room temp.  With the materials that we
have recieved lately, we have tried every combination of solutions from
0.25% up to 5% with times ranging from 5 min to 30 min, but with very poor
results, when compared to  the same control slide that we simply "spit on"
We have far too many slides that require digestion,  on a daily basis to
continue with the  good old "spit" technology.  Has any one out in
histoland got a good source of diastase that works as well as good old
"spit"   Many thanks in advance.
Kerry Beebe
Kelowna Gen Hospital
Kelowna B.C.
Canada
e-mail  bbracing@silk.net




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