[Histonet] soften formalin fixed samples

Tue Jul 27 12:24:20 CDT 2004

Terry,

Here are some things to perhaps look at.

Check on how frequently the waxes have been changed. ( may not have really
good clean wax to infiltrate)

Is there by any chance water in the waxes or in the Xylenes. (the presence
of water in either of these two areas can cause this )

Also check on the frequency of the Xylene changes. ( may not have really
good clean Xylene  to adequately clear out the Alcohol)

Best regards,

Mark

Mark Lewis
Product Specialist
Anatomical Pathology
Clinical Diagnostics
Thermo Electron Corporation
(412) 747-4013
(412) 788-1097
E-mail: mark.lewis <@t> thermo.com

                      "Marshall Terry Dr,                                                                                                           
                      Consultant Histopathologist"          To:       "Gayle Callis" <gcallis <@t> montana.edu>, "Leclerc Jocelyne"                      
                      <Terry.Marshall <@t> rothgen.nhs.uk         <leclercj <@t> vetanat.unizh.ch>, <Histonet <@t> lists.utsouthwestern.edu>                       
                      >                                     cc:                                                                                     
                      Sent by:                              Subject:  RE: [Histonet] soften formalin fixed samples                                  
                      histonet-bounces <@t> lists.utsouth                                                                                                
                      western.edu                                                                                                                   

                      07/27/2004 11:26 AM                                                                                                           

Gayle,

We are having trouble at the moment with poor clearing, giving a grey cast,
and sometimes a steely opaque look to things. Do you think that actually
reducing processing times in alcohol and xylene may help, or is this as
absurd as it sounds?

Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path
 Consultant Pathologist
 Rotherham General Hospital
 South Yorkshire
 England
        terry.marshall <@t> rothgen.nhs.uk

-----Original Message-----
From: Gayle Callis [mailto:gcallis <@t> montana.edu]
Sent: 27 July 2004 16:13
To: Leclerc Jocelyne; Histonet <@t> lists.utsouthwestern.edu
Subject: Re: [Histonet] soften formalin fixed samples

First,  check you tissue processing schedule, it may be too long -
overexposure to alcohols removes too much bound water (bound to proteins)
rather than just free water, over exposure to xylene also hardens tissue,
and heat of paraffin is very drying.  If you samples are very tiny, you can

have shorter processing schedule to prevent drying out of tissue.

Second, you can face or trim the block, and place it face down on a block
of ice with water on top of ice, or just use ice water.  This has been
discussed recently on Histonet (at great length) - go to archives and do a
search for more hints.

At 03:16 AM 7/27/2004, you wrote:
>Good morning everybody!
>
>I have a problem with my formalin fixed samples: they are just too hard.
Now
>I wonder if there is a possibility to soften or "remoisture" them.
>I would be very pleased if anyone of you felt able to help me.
>
>Thank you and best regards,
>
>Jocelyne
>--
>VetSuisse Fakultät
>Veterinär-Anatomisches Institut
>Winterthurerstr. 260
>8057 Zürich
>
>
>_______________________________________________
>Histonet mailing list
>Histonet <@t> lists.utsouthwestern.edu
>http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Gayle Callis
MT,HT,HTL(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)

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