After asking Jean some other questions, she gave permission for me to share
it with Histonetters.  

Excellent hints on agar method. 

Gayle Callis 

>From: "Mitchell (Jean)" 

>My primary agar use has been for EM and on a few occasions have 
>done the exact same procedure when submitting agar embedded 
>samples for paraffin processing. The agar pellet is obtained in a 
>plastic microfuge tube, and once the agar has jelled sufficiently, 
>the end of the tube is sliced off, and the button of an agar pellet is 
>placed back into fixative (be it glut or formalin).
>
>I no longer do paraffin sectioning at the hospital, but once the 
>sample/agar pellet is obtained it is suggested to wrap in lens paper 
>and process as usual.   The temperature of the paraffin should be 
>below the melting point of the agar.  
>
>The only problem I can think of sectioning wise - if there is some 
>kind of contaminant in the agar or if the specimen sample is so 
>dense in the agar it didn't fully process.  But again EM is and has 
>been my primary use of this technique and we have had minimal 
>problems in this area.
>
>Complicated my day - no.  Just made me think a little more on a 
>Friday afternoon.   Jean
>
>
>> Jean,
>> 
>> Further thoughts on this, do you think their processing of agar
>> supported samples is the problem??? I wonder if you could elaborate on
>> HOW your lab processes agar/sample combinations into paraffin since
>> you have no sectioning problems, although you indicated EM was the
>> primary use of this technic.   It could even be a problem with their
>> paraffin sectioning?? or were you talking about EM sectioning only??? 
>> 
>> 
>> I am curious also, hmmm did I make your day more complicated - sorry!
>> 
>> At any rate, have a nice holiday--
>> 
>> 
>> At 09:28 AM 8/30/02 +0000, you wrote:
>> >We use a 4.0% agar solution in distilled water (refrig. storage, 
>> >heating before use) but mostly for EM specimens ( such as nasal 
>> >cilia swabs, urine samples).  Have used this 4.0% solution in 
>> >regular and microwave processing with excellant results.  Never  have
>> >had a problem with
>> sectioning.
>> >
>> >Jean Mitchell, BS, HT 
>> >University of Wisconsin Hospital and Clinics
>> >Department of Neurology
>> >Madison, WI
>> >
>> >
>> >
>> >> Need some aid. Does anyone use agar to support small tissues before
>> >> embedding? If so, can you share a recipe? I have little experience
>> >> with formulas, how to make, etc. The process has been in place for
>> >> some time here but too often, I hear complaints like "agar is
>> >> difficult to cut today." 3 gms of agar in 100 cc NBF heated. Any
>> >> ideas? Thanks
>> >> 
>> >
>> >
>> >
>> >
>> >
>> >
>> Gayle Callis
>> MT,HT,HTL(ASCP)
>> Research Histopathology Supervisor
>> Veterinary Molecular Biology - Marsh Lab
>> Montana State University - Bozeman
>> 19th and Lincoln St
>> Bozeman MT 59717-3610
>> 
>> 406 994-6367 (lab with voice mail)
>> 406 994-4303 (FAX)
>> 
>> email: gcallis@montana.edu
>
>
>
>
Gayle Callis
MT,HT,HTL(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology - Marsh Lab
Montana State University - Bozeman
19th and Lincoln St
Bozeman MT 59717-3610

406 994-6367 (lab with voice mail)
406 994-4303 (FAX)

email: gcallis@montana.edu