Dear Tony,
Thank you for the fine reply on Michel's Transport Media. We had excellent
results with human renal biopsies destined for immunofluorescence staining.
I don't recall ever exceeding 48 hours in Michels as 72 hours was allowed
per recommendation from the original and Elias's publications. The kidney
morphology didn't suffer excessively from this transport media as seen on
our HE stained frozen sections from Liquid Nitrogen cooled isopentane snap
frozen needle biopsies. My renal pathologist always commented that the
frozen section HE looked very much like his the FFPE HE section. This must
vary from laboratory to laboratory and also what tissue was transported e.g.
skin biopsies versus kidney biopsies. We always were very careful with good
Michels buffer rinses.
As for where to access Michels Transport Media and Michels buffer, we
purchased these from Poly Scientific in much larger volumes at a cheaper
price than Wampole(?)(Zeus). Aliquots were made and distributed to
laboratories although that may not be ideal since busy laboratories may find
this a bit labor intensive. I know of laboratories who make up Michels with
great success.
As for transporting tissue on saline soaked gauze, I can't stress the
importance enough to NEVER let the tissue dry out, and snap freeze it as
soon as it arrives in the laboratory. I would prefer receiving a tissue in
cell culture fluid to ensure no drying.
The reference is much appreciated.
G'day and missed seeing you at NSH symposium/convention this year.
Gayle M. Callis
HTL/HT/MT(ASCP)
-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Tony Henwood
Sent: Tuesday, October 05, 2010 5:44 PM
To: Della Speranza, Vinnie; kristen arvidson; histonet
Subject: RE: [Histonet] DIF Transport Media
Vinnie,
I hope you are well.
The following might be of use:
Specimens for immunofluorescence are usually submitted to the laboratory in
cell culture fluid (eg Hanks or RPMI) or on saline soaked gauze. For
transport to other institutions, Michel's Transport Medium has often been
used:
Michel's Buffer
1M potassium citrate, pH 7.02.5 ml
0.1M magnesium sulphate 5.0 ml
0.1M N ethylmalemide5.0 ml
Distilled H2O 87.5 ml
* Mix well and store at 2 8oC. Exp. 1 year
Michel's Transport Medium
Michel's Buffer 100ml
Ammonium sulphate 55gm
Adjust pH to 7.0 7.4 with 1M KOH. Store at 2 8oC. Exp. 1 year
Unfortunately Michel's Transport Medium has erroneously been called Michel's
Fixative. None of the components of Michel's Transport Medium is a fixative.
Ammonium sulphate precipitates antigen-antibody complexes in diseased skin
and renal tissues. N ethylmalemide modifies free sulphydryl groups of
cysteine residues in proteins (Fischer 2006).
Michel's Transport Medium has been shown to be deleterious to morphology.
Ultrastructurally, complete destruction of plasma membranes and
intracytoplasmic organelles occurs after 48 hours storage. On the other
hand, antigenicity is well preserved even after many days storage (Fischer
2006).
Fischer (2006) Intern J Surg Pathol 14(1):108.
Regards
Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC)
Laboratory Manager Senior Scientist
Tel: 612 9845 3306
Fax: 612 9845 3318
the children's hospital at westmead
Cnr Hawkesbury Road and Hainsworth Street, Westmead
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA
-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Della
Speranza, Vinnie
Sent: Wednesday, 6 October 2010 6:10 AM
To: 'kristen arvidson'; histonet
Subject: RE: [Histonet] DIF Transport Media
Zeus was a company that used to market michel's under their own label. I
don't believe zeus still exists. Michel's is the name associated with the
author of the original paper. I don't have the reference.
This solution does not require refrigeration.
Vinnie Della Speranza
Manager for Anatomic Pathology Services
Medical University of South Carolina
165 Ashley Avenue Suite 309
Charleston, South Carolina 29425
Tel: (843) 792-6353
Fax: (843) 792-8974
-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of kristen
arvidson
Sent: Monday, October 04, 2010 2:30 PM
To: histonet
Subject: [Histonet] DIF Transport Media
Is Michel Medium the same as Zeus? Do these need to be refrigerated?
Thanks.
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