Kathleen
We use Michel's regularly, and we rinse for about 15 mins in 10% sucrose in PBS prior to freezing.
Regards Mike

-----Original Message-----
From: Kathleen A Spencer [mailto:KSPENCER@UTMEM.EDU]
Sent: Thursday, 10 April 2003 4:19 AM
To: Behnaz Sohrab; Teresa Flores
Cc: histonet
Subject: RE: ZEUS FIXATIVE

I once worked in a lab where we received renal biopsy's in Michel's
transport medium, but before we could freeze them for cryo-sectioning we
had to take them through several rinses in Michel's rinse or was it called
wash buffer? I can't remember. Does anyone have a clue? I think this is a
very necessary step.

Kathleen Spencer


>===== Original Message From Teresa Flores 
=====
>Behnaz, our lab use to order Zeus Fixative, but the cost and date of
>expiration was always a problem. Below is what our EM lab prepares for
>immunofluorescent studies on renal biopsy, and gives out to our clientel.
>Hope this helps. Teresa
>
>Reference to Michels Transport Media (alias Zeus Fixative):
>Michel M.D., Yorman; Niler Phd, David, Kathy M.T.:  Preservation of tissue
>fixed immunoglobulins in skin biopsies of patients with lupus. J. Invest.
>Derm. 59:449-452 (1973).
>
>CALCULATIONS:
>1.  Prepare a solution of 0.1M K citrate and ph to 7.2 - ph 7.4
>       K-citrate 32.44 grams
>       Distilled water 100.00 ml
>2.  Prepare a solution of 0.1M MgS04 7H2
>       MgS04 7H2 2.40 grams
>       Distilled water 100.00 ml
>3.  Prepare a solution of 0.1M N-ethyl maleimide
>       N-ethyl maleimide 1.2521 grams
>       Distilled water 100.00 ml
>4. Prepare a BUFFER Solution
>               0.1M K-citrate          2.5ml
>               0.1M MgS04 7H2          5.0ml
>               0.1M N-ethyl maleimide  5.0ml
>               Distilled water to make 100.00 ml
>5. The prepared Michel's Transport Media will ph to 7.4
>               (NH4)S04 55 grams
>               Prepared BUFFER Solution 100.00 ml
>A low frequency of false negative will result if biopsy is held in Michel's
>Transport Media for over 5 days.
>
>I WAS WONDERING IF ANY ONE HAS THE MAGIC FORMULA FOR THE " 
ZEUS " FIXATIVE
>. WE USE IT TO TRANSPORT TISSUE. THANKS