[Histonet] Michel's transport medium

[Tom McNemar]

I would think that whoever you send them to should supply the necessary =
transport medias.  At least that's the case with everything I send out.

Tom McNemar, HT(ASCP)
Histology Co-ordinator
Licking Memorial Health Systems
(740) 348-4163
(740) 348-4166
tmcnemar <@t> lmhealth.org
www.LMHealth.org


-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu =
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu]On Behalf Of =
springrosycloud <@t> 126.com
Sent: Wednesday, January 31, 2007 7:40 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Michel's transport medium



Hello, everyone. I am going to use Michel's transport medium to send =
renal tissue samples. I got a protocol on the medium preparations(see =
belows). When I made the preparation according to the protocol, I met =
troubles. At the first step(1.0 M potassium citrate buffer pH 7.0), I =
could not adjust pH using the 1M potassium hydroxide. I had to adjust pH =
to 7.0 using 10M potassium hydroxide(40ml). And then, I could not =
dissolve 55 grams of ammonium sulfate in 100 ml washing solution. About =
half of ammonium sulfate(made in China) kept unsolved, even though I =
incubated it in heat water and/or mechanical stirring. When I bought a =
bottle of new ammonium sulfate, it still did not work. Later, I tried to =
dissolve 55g ammonium sulfate in 100ml distilled water, but I failed. I =
tried and thought it again and again. I did not know what is the matter. =
Please tell me what I should do. I am waiting for your kindly reply.     =
                                      Sincerely,                    =20
                        Betty                                      =20
Nanjing University School of Medicine,=20
 Research institute of Nephrology, Jinling Hospital,=20
 Nanjing, China 210002
Tel: 86-025-81765894
Fax: 86-025-84801992The protocol from =
http://publish.uwo.ca/~jkiernan/faqlist.htm#MICHELammonium sulfate was =
unsolved. 1.0 M potassium citrate buffer pH 7.0:      dissolve 21.0 g =
citric acid monohydrate      (or 19.2 g citric acid anhydrous)      in =
30 mL of hot deionized or distilled water. Cool.    Adjust pH to 7.0 =
with 1 M potassium hydroxide (about 35 mL)    Dilute to 100 mL with more =
water.   Washing solution:      25 mL 1.0 M potassium citrate buffer     =
 50 mL 0.1 M magnesium sulfate heptahydrate (F.W. 246.5)      50 mL 0.1 =
M N-ethyl maleimide (=3D 12.5 g in 1 L of water)                  (Sigma =
E3876.)      Water to make 1 L      Adjust to pH 7.0 with 1 M potassium =
hydroxide      Store in refrigerator. (Cost about $50/25 g in 1994.)   =
Transport medium:      Dissolve 55 grams of ammonium sulfate in 100 mL =
washing        solution. (Add slowly, with mechanical stirring.)      =
Adjust pH to about 6.9 with 1 M potassium hydroxide (< 2 mL =
needed)Reference: Michel B. Milner Y.=20
 David K. Preservation of tissue-fixed immunoglobulins in skin biopsies =
of patients with lupus erythematosus and bullous diseases. A preliminary =
report. J. Invest. Dermatol. 59: 449-452 (1973).
=20
=20
=20
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