Re: [Histonet] Trichrome Question

From:John Kiernan




   Dear  Angela Fogg,
 
It is difficult to= reply without knowing what's in the "Chromaview Stain kit Gomori'= s Trichrome." How does this differ from the method published = by Gomori in Am. J. Clin. Path. 20: 662-664 (1950)?=
 
John Kiernan
Anatomy, UWO
Lon= don, Canada
= = =
----- Original Message -----From: ANGELAFOGG@aol.com
Date: Saturday, November 8= , 2008 10:03
Subject: Re: [Histonet] Trichrome Questi on
To: jkiernan@uwo.ca

>
&g= t;


>
> = =
> =


> Thanks for responding.  Fixed in 10% f= ormalin for 3 weeks, processed
> with formalin, alcoh= olic formalin, alcohols and Clear Rite III for Richard
>= Allan, embedded in EM400 paraffin from Surgipath. Cut at 5 microns = and stained
> using Richard Allan's Chromaview Stain ki= t Gomori's Trichrome.  Used the
> above meth= od on our routine Trichrome controls and all the muscle stained red as < BR>> they are supposed to be.  That's why I put ou= t the question.  The blue
> staining of the mu= scle in the colon puzzled me.  All patient tissue has
= > stained correctly as well.


> Thought ma= ybe the length of time in formalin had something to do with the
= > blue staining.  Maybe longer than an hour in Bouin'= s before continuing the
> Trichrome would help?

> Await your thoughts and info

> Re= gards

> Angela

>  <= BR>
>
> In a message dated 11/8/2008 2:= 10:03 A.M. Eastern Standard Time,
> jkiernan@uw= o.ca writes:

> =
Which
&= gt; trichrome method? What kind of section (frozen, paraffin, pl astic?)
> Any
> post-fixation in somethin= g other than plain formalin before staining?
>
= > Provide the technical details, and you will get lots of advic= e!
>
>  
> Weeks in= formaldehyde is not "prolonged" fixation unless the
>= weeks
> are numerous enough to make years. As a genera lity, trichrome
> methods
> do not work = very well after fixation in liquids that have
> formaldeh= yde as the
> only active ingredient. Postfixation of th= e sections
> can compensate. Bouin
> is = frequently used. Saturated aqueous picric acid us
> jus= t as good. There are
> published reports that iodine and= even citrate
> buffer will improve trichrome
&g= t; staining of paraffin sections of
> formaldehyde-fixe= d tissue. See Yu &
> Chapman 2003 J. =
> Histotechnol.
26(2)= : 131-134.
>
>  
>= ; "Trichrome" has been applied to several staini ng
> techniques that use two
> or more dye= s. By convention, since about 1920,
> trichrome met= hods have
> been those using phosphomolybdic or phosphotun= gstic
> acid (or both) to
> enable the sta= ining of collagen and cytoplasm by
> anionic dye= s with sharply
> contrasting colours: blue or = green for
> collagen, and red for cytoplasm
&g= t; (including smooth & striated muscle). A
> third anionic dye, typically yellow
> or orange, may = be added to stain red
> blood cells. Instructions for <= BR>> trichrome methods can be found in all
> tex= tbooks of  microtechnique and
> histotechnology.=
>  
> John
> Kiern an
> Anatomy, UWO
> London, Canada
= > = = =
> ----- Original Message
&g= t; -----
> From: angelafogg@aol.com
>= Date: Friday, November 7, 2008
> 20:17
&= gt; Subject: [Histonet] Trichrome Question
> To:
> histonet@lists.utsouthwestern.edu
>
> >
> > Performed a trichro= me
> stain on a piece of colon which had been
&= gt; > in formalin for? weeks.?
> Muscle st= ained blue instead of
> > red.?What happened?= ? Does prolonged
> fixation react this way?
&= gt; > Hope someone can shed some light on
> this .
> > Regards,
> > Angela<= BR>> >
> ___________ _______________________ 5F= ____________
> > Histonet m= ailing
> list
> > Histonet@lists.= utsouthwestern.edu
> >
> http:// lists.utsouthwestern.edu/mailman/listinfo/histonet
> < /FONT>


>


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