[Histonet] Buffer.

[Ian Montgomery]

Bonnie,
	You hate ATPase, meet your friend, bane of my life, one of the most
capricious techniques ever to punish a poor Histotech. Although I'm doing a
large number of subjects I keep the numbers down and like your friend 8-9
slides per batch. 9.4 and 4.3 are always ok it's the 4.6 that gives the
grief. 
Ian. 

Dr. Ian Montgomery,
Histotechnology,
I.B.L.S. Support Unit,
Thomson Building,
University of Glasgow,
Glasgow,
G12 8QQ.

-----Original Message-----
From: Whitaker, Bonnie [mailto:Bonnie.Whitaker <@t> osumc.edu] 
Sent: 23 April 2009 16:33
To: ian.montgomery <@t> bio.gla.ac.uk
Subject: RE: [Histonet] Buffer.

Ian,

I hate ATPase, but when I was at University of TX, we did them.  One of my
techs noticed that she did not have as good luck with large batches.  I
can't
really say if this was the case, but she swore they were much better if she
broke them down into smaller batches.  I think she didn't like to do more
than 8-9 slides per batch.

Good Luck!!
Bonnie Whitaker

Clinical Histology Manager
Ohio State University Medical Center
N308B Doan Hall
410 W. 10th Ave.
Columbus, OH  43210

Bonnie.Whitaker <@t> osumc.edu

phone 614.293.5048
fax 614.293.7273
pager 614.346.5013
 

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Ian
Montgomery
Sent: Thursday, April 23, 2009 8:12 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Buffer.

            At the moment I'm doing a large batch of myosin ATPase staining
on human skeletal muscle. 9.4 and the reversal at 4.3 are excellent but the
4.6 is not so good. I will alter the reversal pH in a range from 4.5 to 4.7
but I'm wondering, does the buffer have an effect. I use a glycine buffer
and
have done for many years with good results but this time, mmm, not so good.
Others have used acetate or barbitone and I have myself but always found
glycine to give clean staining after the sulphide rinse.

            Comments, stick to glycine but alter the reversal pH, or change
the whole system and try another buffer?

Ian.     

 

Dr. Ian Montgomery,

Histotechnology,

I.B.L.S. Support Unit,

Thomson Building,

University of Glasgow,

Glasgow,

G12 8QQ.

 

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