Re: [Histonet] copper stain for liver biopsies

From:"Roxanne Soto"

I beg to differ with you John------

I use rhodanine as part of the lindquist method and it works beautifully.  I think it is a matter of working it through until you get the results ---after going through several very expensive control slides I have it down to a perfect slide in a few hours instead of a few days.

  ----- Original Message ----- 
  From: John Kiernan 
  To: Vickroy, Jim 
  Sent: Wednesday, March 23, 2005 5:42 PM
  Subject: Re: [Histonet] copper stain for liver biopsies

  The reagent is p-dimethylaminobenzylidenerhodanine
  (pDMBR). Don't buy rhodanine; it's a different 
  compound and it won't work. Also don't confuse with
  rhodamine and similarly named substances, which are
  completely unrelated.

  pDMBR        24 mg
  100% ethanol  6 ml

  Before using, filter this solution into 45 ml of
  0.065M sodium acetate (= 1.6% anhydrous, or
  2% trihydrate)

  Put hydrated slides in the working solution
  either overnight at 37C or 11 min at 80C.

  Wash X6 in distilled water. Counterstain nuclei
  lightly with haemalum. Wash. Aqueous mountant.
  Cu-containing deposits brownish red.

  Sometimes it's necessary to release "masked"
  copper before doing the method. Before doing
  a histochemical method of this kind, read up 
  about its mechanism, limitations and relations 
  to other methods:

  Pearse's Histochemistry 4th ed Vol 2 (the
  2nd or 3rd ed also OK). 

  C. Chrukian's "Manual of the Special Stains
  Laboratory" 1997 or later edition.

  Irons et al 1977 Arch Path Lab Med 101,298-301
  (This paper compares different methods for Cu.)
  John A. Kiernan
  Department of Anatomy and Cell Biology
  The University of Western Ontario
  London,   Canada   N6A 5C1
  "Vickroy, Jim" wrote:
  > We are looking for a procedure for staining for copper.  In the past we
  > have used an aldehyde fuchsin stain but our pathologist would like to
  > use a rhodanine stain.  Does anybody have a good procedure for staining
  > copper and any suggestions where to get a kit or reagents?
  > Jim Vickroy
  > Memorial Medical Stain
  > Springfield, Illinois
  > _______________________________________________

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