[Histonet] Zues-Michel fixative

[Patti Loykasek]

Hi Joseph. Here is our formula for IF transport media ­ otherwise known as
Zeus or Michel¹s or... I just copied & pasted from our reagent prep. Note
that this reagent does have a shelf life. I¹d be happy to answer any further
questions on this reagent.

Patti Loykasek
PhenoPath Laboratories

PURPOSE

Transport media is used as a holding media for tissue biopsies.  Transport
Media prevents degradation of tissue and immune complexes by inhibiting
tissue enzymes. Tissue can be held in transport media up to 2 weeks without
harm to the tissue.  The various components of the buffer and media have the
following effects
Ammonium sulfate precipitates macromolecules including Antigen-Antibody
complexes without loss of antigenicity, while proteolytic and other
depolymerizing enzymes would be precipitated and inhibited.  A role for such
enzymes in dermal-epidermal separation has been postulated.
Citrate buffer is used for washing because it is least likely to facilitate
dermal-epidermal separation.
Magnesium sulfate neutralizes the chelating capacity of the citrate.
n-Ethylmaleimide minimizes proteolytic activity.

.REAGENTS

1.    Ammonium Sulfate, granular (NH4)2SO4  (VWR, Catalog No. MK351206 or
equivalent). HMIS: 2-0-0

2.    Deionized Water, NCCLS type I

3.    1.0M Potassium Hydroxide HMIS:3-0-2

4.    Transport Buffer HMIS: 2-0-0
PROCEDURE

1.    Transfer 250mL of transport buffer to a large container.
2.    Weigh 137.5g Ammonium Sulfate and transfer to 250mL transport buffer
3.    Mix the solution well and adjust the pH to 7.2 to 7.5 with 1M
Potassium Hydroxide (1M KOH).
Note: The higher pH will extend the shelf life.  Transport media with a pH
below 7.0 and above 7.5 will give unsatisfactory results
4.    Label secondary container with the following information: Name, pH,
Date Prepared, Storage Conditions, Expiration Date (6 months), HMIS 2-0-0,
and initials.
5.    Transport Media is stored at room temperature with a shelf life of 6
months. 

 

REFERENCE DOCUMENTS

1.    HazCom Training Handbook

2.    Material Safety Data Sheet (MSDS)

3.    RP057, 1.0M Potassium Hydroxide Stock Solution

4.    RP041, Transport Buffer from Stock Solutions

5.    1973 Journal of Investigative Dermatology Vol59. No. 6 ³Preservation
of tissue fixed immunoglobins in Skin Biopies of patients with Lupus
Erythemalosis and bullous diseases.  Prelim report Authors (Micheal Miller
and Davied

6.    Immunohistochemistopathology, A Practical Approach to Diagnosis. Jules
M. Elias, 1990, ASCP Press

7.    Chemical SOP

 

 

 

 

 

 

 

 

 

 

SAFETY CAUTION: Ammonium sulfate can be an allergen and an irritant.
Transport buffer is toxic and can be an irritant. Potassium Hydroxide is
caustic and should be stored in a base cabinet.

 

 

PROCEDURE

1.    Transfer 250mL of transport buffer to a large container.

 

2.    Weigh 137.5g Ammonium Sulfate and transfer to 250mL transport buffer

 

3.    Mix the solution well and adjust the pH to 7.2 to 7.5 with 1M
Potassium Hydroxide (1M KOH). Note: The higher pH will extend the shelf
life.  Transport media with a pH below 7.0 and above 7.5 will give
unsatisfactory results

 

4.    Label secondary container with the following information: Name, pH,
Date Prepared, Storage Conditions, Expiration Date (6 months), HMIS 2-0-0,
and initials.

 

5.    Transport Media is stored at room temperature with a shelf life of 6
months. 

 

 

 

 

 





>  Hello all
> 
>   Is anyone out there is in  Histonet can shed any like on Zues- Michel
>  fixative?
> 
>  I  have   a renal biopsy for immunofluorescence, and this fixative was
>  recommended.
> 
>  Thank you all
> 
>   Erskine
>    _________________________________________________________________
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