[Histonet] Histo gel Issues

[Jo Dee Fish]

Hi Jennifer,
I always submerge my cassettes in molten paraffin until I embed them, so
that isn't the problem.  The histogel comes out of the processor hardened
and whitish in appearance.  I also melted the histogel tubes in hot water
while preembedding the embryos, so that probably isn't the problem, either.

Jo Dee 
 

~~Jo Dee Fish~~

Senior Research Technologist

The J. David Gladstone Institutes

Co-manager Histology and Microscopy Core

 

Telephone: (415) 734-2567

Fax: (415) 355-0824

E-mail:  <mailto:jfish <@t> gladstone.ucsf.edu> jfish <@t> gladstone.ucsf.edu

 

Mailing address:

The J. David Gladstone Institutes

1650 Owens Street

San Francisco, CA 94158

 

  _____  

From: Hofecker, Jennifer L [mailto:jennifer.l.hofecker <@t> Vanderbilt.Edu] 
Sent: Tuesday, February 03, 2009 10:37 AM
To: jfish <@t> gladstone.ucsf.edu; pam plumlee; histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] Histo gel Issues



Hi,

Do either of you "dry embed" your Histogel blocks? By this I mean: Are you
submerging the blocks in molten paraffin prior to embedding? Are they coming
off the processor looking different, or are you finding it after you embed
the gel blocks? Does it seem that the gel square is hard and never becomes
“one with the paraffin”? If it’s after embedding, it may be dry embedding
related.

I use Histogel on a daily basis for clinical neurosurgery specimens and have
had no trouble. I do submerge my blocks in molten paraffin after processing
(holding tank of embedding center). I know several people who do not
submerge and they had issues with the gel seeming hard and brittle. If you
dry embed, you can get some good ones, but the likelihood of dried out
"squares" is tremendous. 

Another suggestion may be the temperature of your Histogel. We once had an
overzealous person heat the Histogel so hot that it was smoking (not the
nice happy 50ºC that we’re used to). The block he made before I ‘caught’ him
was hard and brittle the next day while the others were not. I could notice
this difference immediately upon taking them off the processor. 

I must also say that I have not purchased Histogel lately since I buy in
large batches. If there’s been a formula change or a lot related problem, it
would not be affecting me yet. 

Good Luck and please let me know if I can be of any further assistance.

Have a great week!

Jennifer

 

Jennifer L. Hofecker HT(ASCP)

Vanderbilt University Medical Center

Division of Neuropathology

Nashville, TN 

ph 615.343.0083

fax 615.343.7089

 

-----Original Message-----
From: Jo Dee Fish [mailto:jfish <@t> gladstone.ucsf.edu] 
Sent: Tuesday, February 03, 2009 10:50 AM
To: 'pam plumlee'; histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] Histo gel Issues

 

Dear Pam,

I've had the same problem.  I called Richard-Allen just after they were

"absorbed" by Thermo Fisher and got no answers.  They had never heard of

such a problem and didn't know how to solve it.  I have heard from another

user that had the same exact problem.  I lost precious samples, E6.5 mouse

embryos, because of this "drying out and hardening" of the histogel.  I had

to stop using it all together.

If anyone has any suggestions, please let us know!

Take care Pam and all histonetters,

Jo Dee 

 

 

~~Jo Dee Fish~~

Senior Research Technologist

The J. David Gladstone Institutes

Co-manager Histology and Microscopy Core

 

Telephone: (415) 734-2567

Fax: (415) 355-0824

E-mail: jfish <@t> gladstone.ucsf.edu

 

Mailing address:

The J. David Gladstone Institutes

1650 Owens Street

San Francisco, CA 94158

 

-----Original Message-----

From: histonet-bounces <@t> lists.utsouthwestern.edu

[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of pam plumlee

Sent: Monday, February 02, 2009 1:06 PM

To: histonet <@t> lists.utsouthwestern.edu

Subject: [Histonet] Histo gel Issues

 

 

Dear Group: I'm having problems with the processing of small tissues in

histogel. I follow the suggested embedding directions on the package and

then process the gel blocks.  The results are very inconsistent-ideally,

I'll get nice soft gel blocks-but, usually in a batch of 10 I get 2 good

blocks and 8 dried up, flat hard squares.  They are all handled and

processed the same.  Anyone experience this before?  Thanks for any input.

 

Pam Plumlee H.T.

Pfizer La Jolla

pam.plumlee <@t> pfizer.com

 

 

      

 

_______________________________________________

Histonet mailing list

Histonet <@t> lists.utsouthwestern.edu

http://lists.utsouthwestern.edu/mailman/listinfo/histonet