[Histonet] Gomori Green Muscle trichrome.

Sat Aug 13 16:14:55 CDT 2005

Gomori "Green" muscle trichrome: I run a high volume 
IHC/Neuro/Muscle/Bone Marrow lab in VA and we had issues every once in 
a while with the red blotches appearing from Sigma's Gomori Trichrome 
solution. It sounds like witchcraft, but when the solution was agitated 
or mixed or dropped (oops) or freshly poured we noticed a greater 
likelihood of red artefactual staining. (nondescript blotchy red 
patches). We cut muscle frozen in liquid isopentane (n-methyl butane) 
cooled in liquid N2 at 4 microns, we do not fix at all, we do sometimes 
store slides at -70C before staining which does dessicate them, but we 
haven't linked this treatment to the spurious red blotches. try keeping 
your solution in a coplin jar, and don't agitate before staining. We 
pre counterstain in Mayer's Hamatoxylin from Biogenex from our IHC 
proc. for 3- 5 min. Rinse dh2o, gomori one step for 20 minutes, rinse 
tap h2o, aquamount.
On Aug 13, 2005, at 1:01 PM, histonet-request <@t> lists.utsouthwestern.edu 
wrote:

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> Today's Topics:
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>    1. Re: Immunofluorescence problem-Non-specific binding   appears
>       in control slide (al.floyd <@t> juno.com)
>    2. RE: HT Workforce (Lee & Peggy Wenk)
>    3. RE: VIP cleaning cycle (Anne Van Binsbergen)
>    4. more about the gomori's on Frozen muscle (Hofecker, Jennifer L)
>    5. RE: more about the gomori's on Frozen muscle (Anne Van 
> Binsbergen)
>
>
> ----------------------------------------------------------------------
>
> Message: 1
> Date: Fri, 12 Aug 2005 20:56:24 -0400
> From: al.floyd <@t> juno.com
> Subject: Re: [Histonet] Immunofluorescence problem-Non-specific
>     binding appears in control slide
> To: NSTYLOPOULOS <@t> PARTNERS.ORG
> Cc: histonet <@t> lists.utsouthwestern.edu
> Message-ID: <20050812.205624.2372.0.al.floyd <@t> juno.com>
> Content-Type: text/plain; charset=us-ascii
>
> Hello Nick,
>
> You may be running into the problem of Falck-Hillarp reactive cells, 
> more
> commonly referred to as "SIF" cells.  SIF stands for Small Intensely
> Fluorescent cells.  This technique was quite popular back in the 
> 1960's.
> In brief, any cell that contains epinephrine, nor-epi, or Dopamine will
> become very highly fluorescent after exposure to formalin.  In the 
> actual
> Falck-Hillarp technique, the exposure is to gaseous formaldehyde vapor,
> but you may be getting some of the same reaction with your procedure.
> Generally, Epi and Nor-epi fluoresce with a color that is very similar 
> to
> flourescein, while Dopamine has a more orange color.
>
> There is actually a monograph available titled SIF cells.  I forget the
> exact date, but think it is early '70's.  I have a copy on my 
> bookshelf,
> but at the moment am a long way from the bookshelf!
>
> Try a Google, Google Scholar or PubMed search for SIF cells - you 
> should
> find quite a few publications.  It was a very popular technique before
> the days of immunostains.
>
> Al Floyd
> Alton D. Floyd, Ph.D.
> 23126 South Shore Drive
> Edwardsburg, MI 49112
> Cell: 574 215-0703
>
>
>
> ------------------------------
>
> Message: 2
> Date: Sat, 13 Aug 2005 01:01:11 -0400
> From: "Lee & Peggy Wenk" <lpwenk <@t> sbcglobal.net>
> Subject: RE: [Histonet] HT Workforce
> To: "'Janice A Mahoney'" <JMahoney <@t> alegent.org>
> Cc: 'Histonet submissions' <histonet <@t> lists.utsouthwestern.edu>
> Message-ID:
>     <mailman.0.1123952400.5979.histonet <@t> lists.utsouthwestern.edu>
> Content-Type: text/plain;   charset="US-ASCII"
>
> Information available about the laboratory workforce in general:
>
> http://www.labmedicine.com/headlines/ascpnews/040205.html
> 72% of laboratory workers are older than the age of 40. Approximately 
> 30%
> are older than the age of 50
>
> Peggy A. Wenk, HTL(ASCP)SLS
> William Beaumont Hospital
> Royal Oak, MI 48073
>
>
>
> Lee & Peggy Wenk
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu
> [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Janice 
> A
> Mahoney
> Sent: Thursday, August 11, 2005 4:49 PM
> To: histonet <@t> lists.utsouthwestern.edu
> Subject: [Histonet] HT Workforce
>
> Hello,
> Does anyone know what the average age of registered Histo techs is?  
> How
> many retire each year and how many new ones come into the field?
> Thanks for the help.
> Jan
> Omaha
>
> Janice Mahoney
> Histology/Cytology Coordinator
> Alegent Health Laboratory
> Phone (402)717-2889
> Fax (402)717-5231
>
> "We pledge to be creative, visionary leaders committed to holistic
> healthcare in the region"
>
>
>
>
>
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>
>
>
> ------------------------------
>
> Message: 3
> Date: Sat, 13 Aug 2005 11:35:49 +0400
> From: "Anne Van Binsbergen" <vanann702 <@t> skmc.gov.ae>
> Subject: RE: [Histonet] VIP cleaning cycle
> To: "Fred Underwood" <funderwood <@t> mcohio.org>,
>     <histonet <@t> lists.utsouthwestern.edu>
> Message-ID:
>     <0C44F1AAEE47D54DA4210A60AB206F5E02F6D5CD <@t> SKMCEMAIL.skmc.gov.ae>
> Content-Type: text/plain;   charset="us-ascii"
>
> My Vip5 does just fine on a 7 day cleaning cycle. I use the shortened
> cleaning cycle for 6 days and do a long one once in the 7 day period. I
> am also fastidious about having a hot water flush once a month on the
> first 4 stations. My general solutions are 'rotated' every 7 days and
> the wax is as well - discard 1st, shift all up and renew last.
> Annieinarabia (formerly Annieinafrica)
>
> -----Original Message-----
> From: Fred Underwood [mailto:funderwood <@t> mcohio.org]
> Sent: Thursday, August 11, 2005 8:10 PM
> To: histonet <@t> lists.utsouthwestern.edu
> Subject: [Histonet] VIP cleaning cycle
>
> Hi in Hisotland,
>
> This question is for those with a VIP 5 processor.  Has anyone tried to
> get more than the recommended 5 runs from the cleaning xylene and
> alcohol?
>
> Thanks in adavance,
>
> Fred Underwood
> Mont. Co. Coroner
> Dayton, OH
>
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>
>
> ------------------------------
>
> Message: 4
> Date: Sat, 13 Aug 2005 07:53:20 -0500
> From: "Hofecker, Jennifer L" <jennifer.l.hofecker <@t> Vanderbilt.Edu>
> Subject: [Histonet] more about the gomori's on Frozen muscle
> To: "histonet" <histonet <@t> lists.utsouthwestern.edu>
> Message-ID:
>     <898D946569A27444B65667A49C07405207545D <@t> mailbe06.mc.vanderbilt.edu>
> Content-Type: text/plain;   charset="iso-8859-1"
>
> First,
>
> Thank you to those of you that responded. Especially those who 
> understood that when doing modified Gomori's for a neuropathologist, 
> muscle should stain green. I realize that in FFPE, muscle stains red, 
> however our muscle has been staining correctly (green) until this 
> week. I think that over the past several years, the neuropathologist 
> might have noticed if we were staining the muscle the completely wrong 
> color. (sorry, a little sarcastic)
>
> I am also not fixing the sections at all. We go right into 
> hematoxylin. A private reply I received suggested that putting fresh 
> slides in the freezer may be acting as "fixation" (lyophilization) 
> thus, the fresh cuts aren't getting fixed. To those of you doing neuro 
> muscle staining (remember, we're searching for normal to be GREEN) do 
> you use fixation before staining?
>
> I would like to remind some of you that histonet is a tool for us to 
> help each other out. It really is not for use as a weapon. I honestly 
> needed advice, but I as usual, hesitated because I was afraid of the 
> "less than kind" responses that seem to come with many of my posts.
>
> Thanks again to those who were kind enough to help me out. There is 
> such a wealth of knowledge out there in histoland. Hopefully somebody 
> will have the answer.
>
> Bracing for the next round,
>
> Jennifer
>
>
>
> Jennifer Hofecker, HT (ASCP)
>
> Vanderbilt University Medical Center
>
> Division of Neuropathology
>
> (615) 343-0083
>
> (615) 343-7089 fax
>
>
>
> ------------------------------
>
> Message: 5
> Date: Sat, 13 Aug 2005 17:06:44 +0400
> From: "Anne Van Binsbergen" <vanann702 <@t> skmc.gov.ae>
> Subject: RE: [Histonet] more about the gomori's on Frozen muscle
> To: "Hofecker, Jennifer L" <jennifer.l.hofecker <@t> Vanderbilt.Edu>,
>     "histonet" <histonet <@t> lists.utsouthwestern.edu>
> Message-ID:
>     <0C44F1AAEE47D54DA4210A60AB206F5E02F6D5CF <@t> SKMCEMAIL.skmc.gov.ae>
> Content-Type: text/plain;   charset="us-ascii"
>
> Ran a neuropath lab for many years and did muscle bx's galore
> I used to cut my sections and store them at -20.
> the next day, id fix in methanol (I think for 5 mins), rinse in h2o,
> stain in harris' hx, another rinse, into the trichrome stain for 10-15
> mins, quick rinse, few dips in weak acetic acid, DC+M
> Method from Carleton. Used it for many years with great success.
> FYI - this trichrome stain works well on FFPE - colours are a little
> different but it's a 'no hassle' stain.
> Works for me!!
> Hope this helps
> Annieinarabia
>
> -----Original Message-----
> From: Hofecker, Jennifer L [mailto:jennifer.l.hofecker <@t> Vanderbilt.Edu]
> Sent: Saturday, August 13, 2005 4:53 PM
> To: histonet
> Subject: [Histonet] more about the gomori's on Frozen muscle
>
> First,
>
> Thank you to those of you that responded. Especially those who
> understood that when doing modified Gomori's for a neuropathologist,
> muscle should stain green. I realize that in FFPE, muscle stains red,
> however our muscle has been staining correctly (green) until this week.
> I think that over the past several years, the neuropathologist might
> have noticed if we were staining the muscle the completely wrong color.
> (sorry, a little sarcastic)
>
> I am also not fixing the sections at all. We go right into hematoxylin.
> A private reply I received suggested that putting fresh slides in the
> freezer may be acting as "fixation" (lyophilization) thus, the fresh
> cuts aren't getting fixed. To those of you doing neuro muscle staining
> (remember, we're searching for normal to be GREEN) do you use fixation
> before staining?
>
> I would like to remind some of you that histonet is a tool for us to
> help each other out. It really is not for use as a weapon. I honestly
> needed advice, but I as usual, hesitated because I was afraid of the
> "less than kind" responses that seem to come with many of my posts.
>
> Thanks again to those who were kind enough to help me out. There is 
> such
> a wealth of knowledge out there in histoland. Hopefully somebody will
> have the answer.
>
> Bracing for the next round,
>
> Jennifer
>
>
>
> Jennifer Hofecker, HT (ASCP)
>
> Vanderbilt University Medical Center
>
> Division of Neuropathology
>
> (615) 343-0083
>
> (615) 343-7089 fax
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
> ------------------------------
>
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> End of Histonet Digest, Vol 21, Issue 17
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