RE: ZEUS FIXATIVE

From:Teresa Flores

>Date: Wed, 9 Apr 2003 13:14:17 -0500
>To: Kathleen A Spencer 
>From: Teresa Flores 
>Subject: RE: ZEUS FIXATIVE
>Cc:
>Bcc:
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>Yes, Kathleen, sorry I forgot to include that part, it is PBS (Phosphate
>buffered salne ph 7.4) three, 10 minute each change, of PBS. Then you are
>ready to orient in oct and embed, freeze and section. Teresa
>>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
>





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