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 > >
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