Re: [Histonet] Histogel Problem

2011-06-26 Thread dusko trajkovic 
Amos,
I have had the same problem in the past, and posted my issues on the Histonet, 
however no one was able to help me out. At one point I even exchanged Histogel 
with a colleage severl hundred miles away, thinking that maybe I had a bad lot 
of Histogel. She did not have a problem with my Histogel and neither did I with 
her Histogel. The shrinking was arbitrary. At times all of them look great and 
other times more than half shrunk and looked brittle. Even RA/Thermo did not 
have an answer for me.
I decided to do an experiment. To not bore everyone on the Histonet and explain 
all of my experimental steps, what it boiled down to is that you need a long 
processing program on the processor. We use a VIP processor as well, and the 
processing program is at least 12 hours long. NO MORE PROBLEMS. 

Since I started using this progrem, every single Histogel block has been 
perfect.
Let me know if you need any further info or explanation.

Dusko Trajkovic HT ASCP
Pfizer Inc. La Jolla

 




From: Amos Brooks amosbro...@gmail.com
To: histonet@lists.utsouthwestern.edu
Sent: Fri, June 24, 2011 10:29:09 AM
Subject: [Histonet] Histogel Problem

Hi,
  I have a problem with some blocks that were prepared with Histogel. I was
hoping someone else might have had a similar problem and figured it out. I
took a photo of the blocks that were mads and put them in a Picassa album
here:
https://picasaweb.google.com/lh/photo/APO3HsIMa2_jPOEs3QRGUjhz3qi22FNPb2i5JJnBCAk?feat=directlink

  The long  short of it is that the blocs were prepared by the researcher
for me to process. They are mouse kidneys. Now it is entirely possible for
him to have goofed something up in preparing the Histogel blocks. I wasn't
there when he did it, but when I looked at them before processing, they all
looked fine. (Like the adjacent good one in the photo). When they were
processed they were placed in the VIP right next to each other. When I went
to embed them this morning all but one of the four looked fine. The one that
didn't come out well looked like the Histogel had shrunk up and shriveled
around the kidneys. I am sure this will be aweful to cut, and the researcher
is going to have a bird over it since this happened with another project
previously. I would like to have a decent explanation for him, so if anyone
knows what might have happened and has suggestions I would love to hear it
(yes vendors too are welcome to answer this of course!).
  By the way, this was processed on a rather short cycle of 15-20 min per
station of graded ETOH from 70% to 100% with 3 xylene stations and 4
paraffin stations (45 min for these). It seems fine for everything else that
was on the processor. Just that one Histogel block was the issue.

Thank you,
Amos
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RE: [Histonet] Histogel Problem

2011-06-24 Thread Delossantos_Roseann 
Hi,
I've encountered this problem before in my previous lab.  To reduce the 
Histogel from shrinking that badly, avoid putting the tissue at the very edge 
of the Histogel, space them out nicely in the middle, providing sufficient 
amount of extra Histogel between each tissue and surrounding them.  A little 
bit of shrinking usually do not interfere with cutting as long as the whole 
thing is placed flat on the final paraffin block, the rest will stretch out on 
the water bath.  I find sometimes the gel does not behave well in the water but 
as long as it is not on top of your tissue when placed on your slide, it should 
interfere much with staining.


Rose



-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Amos Brooks
Sent: Friday, June 24, 2011 10:29 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Histogel Problem

Hi,
   I have a problem with some blocks that were prepared with Histogel. I was
hoping someone else might have had a similar problem and figured it out. I
took a photo of the blocks that were mads and put them in a Picassa album
here:
https://picasaweb.google.com/lh/photo/APO3HsIMa2_jPOEs3QRGUjhz3qi22FNPb2i5JJnBCAk?feat=directlink
   The long  short of it is that the blocs were prepared by the researcher
for me to process. They are mouse kidneys. Now it is entirely possible for
him to have goofed something up in preparing the Histogel blocks. I wasn't
there when he did it, but when I looked at them before processing, they all
looked fine. (Like the adjacent good one in the photo). When they were
processed they were placed in the VIP right next to each other. When I went
to embed them this morning all but one of the four looked fine. The one that
didn't come out well looked like the Histogel had shrunk up and shriveled
around the kidneys. I am sure this will be aweful to cut, and the researcher
is going to have a bird over it since this happened with another project
previously. I would like to have a decent explanation for him, so if anyone
knows what might have happened and has suggestions I would love to hear it
(yes vendors too are welcome to answer this of course!).
   By the way, this was processed on a rather short cycle of 15-20 min per
station of graded ETOH from 70% to 100% with 3 xylene stations and 4
paraffin stations (45 min for these). It seems fine for everything else that
was on the processor. Just that one Histogel block was the issue.

Thank you,
Amos
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