Fwd: Re: [Histonet] processing fatty tissue

From:Rene J Buesa



Maxim:
  My answer to Robert without the attachment.
  RenÚ J.

Rene J Buesa  wrote: 
  Date: Fri, 1 Jun 2007 05:48:45 -0700 (PDT)
From: Rene J Buesa 
Subject: Re: [Histonet] processing fatty tissue
To: Robert Chiovetti 

  Robert:
  The procedure is attached. The fundament is a GENTLE dehydrant substitution by the infiltrating agent (paraffin) and ELIMINATES the use of an antemedium (xylene).
  The general function of an antemedium is the ability to be the "connection" between the dehydrant and the wax (paraffin) because it mixes with both.
  Xylene (as well as white naphtha or some other aromatic chemicals) can do this, BUT the thing is that mineral oil (MO) IS paraffin of a low molecular weight so the antemedium is not needed or a mixture of MO with alcohols constitutes the antemedium.
  My procedure uses EthOL to dehydrate and later the antemedium is substituted by the mixture of EthOL + Isopropanol + MO
  Maxim has simplified the procedure because he process manually and don't have the advantage of vacuum or pressure or agitation as I did when developed the method.
  So it turns out that Maxim's modification is easier and more direct; he just dehydrates with propanol and later goes into the gentle substitution with a mixture of 5 parts of propanol + 1 part of MO heated at 50║C followed by another mixture of 2 parts of propanol + 1 of MO heated also at ║C to obtain the gentle and complete infiltration of ANY type of tissue.
  The infiltration with MO gives the tissues a softness never achieved with any other antemedium. You will see when you try it.
  Maxim's method is simpler than mine and, in the long run, will be more acceptable to all histotechs and also meets the objective of eliminating xylene from the histology lab.
  The procedure uses 2 chemicals that are cheaper and when needing to be disposed off, the propanol can be evaporated, the used MO mixed with used paraffin and both disposed off as a solid, cutting costs also in disposal.
  Try it, you will like it! 
RenÚ J.

Robert Chiovetti  wrote: 
  Maxim, Rene (and Other Histonetters),

That's interesting re: using either a mix of ethanol+isopropanol+mineral oil (Rene) or isopropanol+mineral oil (Maxim) for breast tissue. Could you share your recipes with us? 

I have a customer (derm path) who could probably benefit from this for larger and thicker skin specimens which sometimes have a lot of subcutaneous fat associated with them.

Thanks in advance, if you could share the recipes!

Cheers,

Bob

Robert (Bob) Chiovetti, Ph.D.
Southwest Precision Instruments
The Desert Southwest's Microscopy Resource
Tucson, Arizona USA
Tel./Fax 520-546-4986
www.swpinet.com
Member, Arizona Small Business Association
(www.asba.com)








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