More on agar from Jean Mitchell
After asking Jean some other questions, she gave permission for me to share
it with Histonetters.
Excellent hints on agar method.
>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
>> 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
>> >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
>> 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: email@example.com
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)
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