[Histonet] modified gomori stain

Sat Jun 17 22:47:35 CDT 2006

In a frozen section, muscle should be green - how else would one demonstrate ragged red fibres???
Anne

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Katri Tuomala
Sent: Saturday, June 17, 2006 9:52 PM
To: manal galal; histonet <@t> lists.utsouthwestern.edu
Subject: Re: [Histonet] modified gomori stain

If you are referring to Gomori's one step trichrome stain, muscle should be 
staining red, not green as collagen. See web page 
http://stainsfile.info/StainsFile/theory/tri_gen.htm

Katri

Katri Tuomala
Hamilton, Ontario, Canada

----- Original Message ----- 
From: "manal galal" <galalmkh <@t> yahoo.com>
To: <histonet <@t> lists.utsouthwestern.edu>
Sent: Saturday, June 17, 2006 4:38 AM
Subject: [Histonet] modified gomori stain

> Hi,
>    I need help with modified gomori  stain. I cant get the muscle to be 
> green. Could anyone help me.
>                                                                 Dr. Manal 
> Galal
>               Consultant Pathology
>                                                               Institute of 
> Neuromotor Disabilities
>                                                             Cairo, Egypt
>
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> Today's Topics:
>
> 1. RE: Trichrome staining and fibrin
> (Marshall Terry Dr, Consultant Histopathologist)
> 2. substrates (Perry, Margaret)
> 3. toe nails (Conlon, Paula F.)
> 4. Re: substrates (Rene J Buesa)
> 5. Re: toe nails (Rene J Buesa)
> 6. Re: toe nails (Chris Pomajzl)
> 7. ScyTek's serum free pro-block protein (Maria Mejia)
> 8. RE: toe nails (Anthony F. Boris)
> 9. Manual cover slip method - DiscoverSlip? (Geoffrey White)
> 10. Re: substrates (Susan Q Wells)
> 11. Sphingomyelin (mtitford <@t> aol.com)
> 12. High profile blades (Snider, Deanna)
> 13. Mechanical testing-Dallas-Fort Worth (Ashwin Nair)
>
>
> ----------------------------------------------------------------------
>
> Message: 1
> Date: Wed, 14 Jun 2006 13:39:24 +0100
> From: "Marshall Terry Dr, Consultant Histopathologist"
>
> Subject: RE: [Histonet] Trichrome staining and fibrin
> To: "Bryan Hewlett" , Timo V?is?nen
> ,
> Message-ID:
>
> Content-Type: text/plain; charset="iso-8859-1"
>
> Well, that's telling me:-)
>
> All these years, the success I've had with primary mercury fixation (I had 
> the original article once), and lack of success with anything other was 
> entirely due to something else, the techs washing too much in one instance 
> and little enough in another.
> Who would have thought it.
> However it may explain my observation that "At times it works and at times 
> not."
>
> Actually, the only time you can be confident that you are looking at 
> fibrin, is when it is in strands - easily recognised in H&E.
> To call anything else fibrin on any tinctorial stain is something akin to 
> a leap of faith.
>
> But - I'll get you back re. your "bees can't fly" thing.
>
> Take this/that: :-)
>
> The "science has proved that bees can't fly" urban myth originated in a 
> 1934 book by entomologist Antoine Magnan, who discussed a mathematical 
> equation by Andre Sainte-Lague, an engineer. The equation proved that the 
> maximum lift for an aircraft's wings could not be achieved at equivalent 
> speeds of a bee. I.e., an airplane the size of a bee, moving as slowly as 
> a bee, could not fly. Although this did not mean a bee can't fly (which 
> after all does not have stationary wings like the posited teensy 
> aircraft),
> nevertheless the idea that Magnan's book said bees oughtn't be able to fly 
> began to spread.
>
>
> Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path
> Consultant Pathologist
> Rotherham General Hospital
> South Yorkshire
> England
> terry.marshall <@t> rothgen.nhs.uk
>
> -----Original Message-----
> From: Bryan Hewlett [mailto:bhewlett <@t> cogeco.ca]
> Sent: 13 June 2006 17:14
> To: Marshall Terry Dr, Consultant Histopathologist; Timo Väisänen;
> histonet <@t> lists.utsouthwestern.edu
> Subject: Re: [Histonet] Trichrome staining and fibrin
>
>
> Terry,
>
> Actually NOT!
> True, Lendrum's original papers(1962) recommended fixation in 
> picro-mercuric
> alcohol for 3 weeks (a tad unnecessary that!).
> However, both Prof. Lendrum and Bill Slidders also successfully used
> formaldehyde fixed material that was treated in Bouin immediately prior to
> staining.
> The key to all of the Masson Type trichromes on formaldehyde-fixed tissue,
> is to use a pretreatment in picric acid.
> This re-aligns the reactive side chains on the proteins, so that there is 
> a
> predominance of basic amino groups and hence maximal binding of anionic
> dyes.
> Mercury fixation is simply not necessary! Nor is Zinc substitution. (see
> Troubleshooting Histology stains: Horobin and Bancroft 1998, page 129)
> The lack of consistency, in obtaining good red colour, is due variability 
> in
> the staining procedure (the post dye water rinses), NOT the fixation.
> Once this is addressed, the inconsistency goes away.
> That is true for the original Masson, Picro-Mallory (all variants), MSB 
> and
> Masson 44/41.
> Achieving consistent good red colour with formaldehyde fixed material is
> easy, if the staining mechanisms are understood and those unnecessary 
> water
> rinses removed.
> Been doing it for over 40 years!
>
> Bryan
> (Engineers can prove that the bumble bee simply cannot fly. However, the
> bumble bee doesn't know that and flies anyway)
>
> ----- Original Message ----- 
> From: "Marshall Terry Dr, Consultant Histopathologist"
>
> To: "Bryan Hewlett" ; "Timo Väisänen"
> ;
> Sent: Tuesday, June 13, 2006 11:27 AM
> Subject: RE: [Histonet] Trichrome staining and fibrin
>
>
> Bryan,
>
> You know full well that MSB required initial staining in mercury. I know
> that zinc is an adequate replacement.
> Attempting to get a consistent good red colour with formalin fixed 
> material
> is futile, as is post-fixation.
> At times it works and at times not.
>
> IMO, Masson is useless for fibrin.
>
> Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path
> Consultant Pathologist
> Rotherham General Hospital
> South Yorkshire
> England
> terry.marshall <@t> rothgen.nhs.uk
>
> -----Original Message-----
> From: Bryan Hewlett [mailto:bhewlett <@t> cogeco.ca]
> Sent: 13 June 2006 15:52
> To: Timo Väisänen; histonet <@t> lists.utsouthwestern.edu
> Subject: Re: [Histonet] Trichrome staining and fibrin
>
>
> Timo,
>
> Try the Lendrum MSB trichrome variant, it was designed to demonstrate
> fibrin.
> I will send the method via a separate e-mail.
>
> Bryan
>
> ----- Original Message ----- 
> From: "Timo Väisänen"
> To:
> Sent: Tuesday, June 13, 2006 10:33 AM
> Subject: [Histonet] Trichrome staining and fibrin
>
>
>> Hello all,
>>
>> I have struggled with getting Masson Trichrome (Goldner) staining to work
>> with
>> kidney biopsies. The problem is that our pathologist does not get 
>> positive
>> fibrin staining to glomeruli in diseased kidneys. Those areas of the
>> glomeruli
>> that should contain fibrin, at least according to the pathologist, are
>> green
>> (Lichtgrun) and not red, as they should be, whatever I do. I have tried 
>> to
>> enhance the staining with Bouin's fixative (+56C) pretreatment but it did
>> not
>> change the overall situation. However, the red stain was more intense in
>> skin
>> samples containing fibrin (formalin fixed). I have also tried another
>> Trichrome
>> staining with Crocein Scarlet 7B without any luck. In our lab kidney
>> biopsies
>> are routinely fixed with alcoholic Bouin's. As far as I know it should be
>> compatible with the Masson protocol we use. Any ideas? Any good protocols
>> that
>> I could try?
>>
>> Thank's in advance,
>>
>> Timo
>>
>>
>> _______________________________________________
>> Histonet mailing list
>> Histonet <@t> lists.utsouthwestern.edu
>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>>
>
>
>
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>
>
> ------------------------------
>
> Message: 2
> Date: Wed, 14 Jun 2006 09:08:36 -0500
> From: "Perry, Margaret"
> Subject: [Histonet] substrates
> To:
> Message-ID:
>
> Content-Type: text/plain; charset="us-ascii"
>
> We are trying to do double staining and are looking for two different
> colors of substrate and we can't use DAB due to melanin. Does anyone
> know of a green stain that is xylene compatible? Vector has Nova Red
> and a blue color but we use hematoxylin as a counterstain. It would
> mean an extra step for us to counterstain in Methyl Green
>
> We use a DAKO autostainer.
>
>
>
> Margaret Perry HT(ASCP)
>
> South Dakota State University
>
> Animal Research and Diagnostic Lab
>
> Brookings SD 57007
>
> Margaret.Perry <@t> sdstate.edu
>
>
>
>
>
> ------------------------------
>
> Message: 3
> Date: Wed, 14 Jun 2006 10:16:10 -0400
> From: "Conlon, Paula F."
>
> Subject: [Histonet] toe nails
> To: "Histonet \(E-mail\)"
> Message-ID:
>
> Content-Type: text/plain; charset="iso-8859-1"
>
> Hi Histonetters, does anyone have a good procedure for sectioning nails 
> and keeping them on the slides during h&e staining and pas staining?Most 
> of the time our sections wash off. Thanks for your help.
>
>
> See our web page at http://www.lahey.org for a full directory of Lahey 
> sites, staff, services and career opportunities.
>
> THIS MESSAGE IS INTENDED FOR THE USE OF THE PERSON TO WHOM IT IS 
> ADDRESSED. IT MAY CONTAIN INFORMATION THAT IS PRIVILEGED, CONFIDENTIAL AND 
> EXEMPT FROM DISCLOSURE UNDER APPLICABLE LAW. If you are not the intended 
> recipient, your use of this message for any purpose is strictly 
> prohibited. If you have received this communication in error, please 
> delete the message and notify the sender so that we may correct our 
> records.
>
>
> ------------------------------
>
> Message: 4
> Date: Wed, 14 Jun 2006 07:23:19 -0700 (PDT)
> From: Rene J Buesa
> Subject: Re: [Histonet] substrates
> To: "Perry, Margaret" ,
> histonet <@t> lists.utsouthwestern.edu
> Message-ID: <20060614142319.7044.qmail <@t> web61216.mail.yahoo.com>
> Content-Type: text/plain; charset=iso-8859-1
>
> Margaret:
> Confronted with that situation I would eliminate the melanine better. 
> Oxidize it with 0.1% potassium permanganate for 2 hours; wash and remove 
> the oxidized melanine with 0.3% oxalic acid followed by washing. The 
> section should be free of melanine pigment/colour afterwards and you can 
> use DAB (for brown) and DAB + Ni salts for dark blue as double substrates.
> Hope this will help you!
> René J.
>
> "Perry, Margaret" wrote:
> We are trying to do double staining and are looking for two different
> colors of substrate and we can't use DAB due to melanin. Does anyone
> know of a green stain that is xylene compatible? Vector has Nova Red
> and a blue color but we use hematoxylin as a counterstain. It would
> mean an extra step for us to counterstain in Methyl Green
>
> We use a DAKO autostainer.
>
>
>
> Margaret Perry HT(ASCP)
>
> South Dakota State University
>
> Animal Research and Diagnostic Lab
>
> Brookings SD 57007
>
> Margaret.Perry <@t> sdstate.edu
>
>
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
>
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> ------------------------------
>
> Message: 5
> Date: Wed, 14 Jun 2006 07:25:13 -0700 (PDT)
> From: Rene J Buesa
> Subject: Re: [Histonet] toe nails
> To: "Conlon, Paula F."
> , "Histonet
> \(E-mail\)"
> Message-ID: <20060614142513.15660.qmail <@t> web61219.mail.yahoo.com>
> Content-Type: text/plain; charset=iso-8859-1
>
> Paula:
> I am forwarding privately a summary on Toenails I prepared recently that I 
> hope will answer your questions.
> René J.
>
> "Conlon, Paula F."
> wrote:
> Hi Histonetters, does anyone have a good procedure for sectioning nails 
> and keeping them on the slides during h&e staining and pas staining?Most 
> of the time our sections wash off. Thanks for your help.
>
>
> See our web page at http://www.lahey.org for a full directory of Lahey 
> sites, staff, services and career opportunities.
>
> THIS MESSAGE IS INTENDED FOR THE USE OF THE PERSON TO WHOM IT IS 
> ADDRESSED. IT MAY CONTAIN INFORMATION THAT IS PRIVILEGED, CONFIDENTIAL AND 
> EXEMPT FROM DISCLOSURE UNDER APPLICABLE LAW. If you are not the intended 
> recipient, your use of this message for any purpose is strictly 
> prohibited. If you have received this communication in error, please 
> delete the message and notify the sender so that we may correct our 
> records.
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
>
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> ------------------------------
>
> Message: 6
> Date: Wed, 14 Jun 2006 09:41:48 -0500
> From: "Chris Pomajzl"
> Subject: Re: [Histonet] toe nails
> To: "HISTONET"
> Message-ID: <002e01c68fc0$b2fb6330$26fca8c0 <@t> CSP>
> Content-Type: text/plain; charset="iso-8859-1"
>
> Good question. We have dealt with this issue for many years, and we have
> come up with a solution that has worked for some time.
>
> First of all, sections are picked up on "+" coated slides that have been
> smeared with egg albumin. We separate eggs from the grocery store and keep 
> a
> stock of the egg white albumin in a bottle. Maybe 100ml at a time, keep it
> refrigerated. When cutting nails, put 2-3 drops of albumin on the slide 
> and
> smear to cover all of the glass.
>
> We cut 3 sections of the nail specimen. 1 for H&E, 1 for PAS, and 1 extra
> for back-up. We bake the slides at 55'C overnight, and then at 70'C for 20
> minutes prior to staining the following day.
>
> This seems to work very well for us.
>
> ----- Original Message ----- 
> From: "Conlon, Paula F."
>
> To: "Histonet (E-mail)"
> Sent: Wednesday, June 14, 2006 9:16 AM
> Subject: [Histonet] toe nails
>
>
> Hi Histonetters, does anyone have a good procedure for sectioning nails 
> and
> keeping them on the slides during h&e staining and pas staining?Most of 
> the
> time our sections wash off. Thanks for your help.
>
>
> See our web page at http://www.lahey.org for a full directory of Lahey
> sites, staff, services and career opportunities.
>
> THIS MESSAGE IS INTENDED FOR THE USE OF THE PERSON TO WHOM IT IS 
> ADDRESSED.
> IT MAY CONTAIN INFORMATION THAT IS PRIVILEGED, CONFIDENTIAL AND EXEMPT 
> FROM
> DISCLOSURE UNDER APPLICABLE LAW. If you are not the intended recipient, 
> your
> use of this message for any purpose is strictly prohibited. If you have
> received this communication in error, please delete the message and notify
> the sender so that we may correct our records.
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
>
> ------------------------------
>
> Message: 7
> Date: Wed, 14 Jun 2006 08:11:41 -0700
> From: Maria Mejia
> Subject: [Histonet] ScyTek's serum free pro-block protein
> To: histonet <@t> lists.utsouthwestern.edu
> Message-ID:
> Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed
>
> To anyone who using ScyTek's serum free pro-block protein, please let
> me know
> what you think of it. In IHC, do you (only) use it as a diluent for
> the antibodies or also
> use it in the washes? Any information you can provide will be greatly
> appreciated.
>
> Yours
>
> Maria Bartola Mejia
> UCSF
> Depart. of Neurosurgery
> San Francisco, CA 94103
>
>
>
> ------------------------------
>
> Message: 8
> Date: Wed, 14 Jun 2006 11:51:21 -0400
> From: "Anthony F. Boris"
> Subject: RE: [Histonet] toe nails
> To: "HISTONET"
> Message-ID:
> Content-Type: text/plain; charset="utf-8"
>
> We use charged slides from Premiere. keep in 65 degree oven for 30 minutes 
> and stain. We have not had any fall off since we switched to these slides. 
> When cutting you can use a 5% solution of KOh for a "surface decal". This 
> breaks down the keratin a little bit (same stuff as in NAIR). Helps to get 
> decent sectioning.
>
> Tony
>
> -----Original Message----- 
> From: Chris Pomajzl [mailto:cpomajzl <@t> cpllabs.com]
> Sent: Wed 6/14/2006 10:41 AM
> To: HISTONET
> Cc:
> Subject: Re: [Histonet] toe nails
>
>
>
> Good question. We have dealt with this issue for many years, and we have
> come up with a solution that has worked for some time.
>
> First of all, sections are picked up on "+" coated slides that have been
> smeared with egg albumin. We separate eggs from the grocery store and keep 
> a
> stock of the egg white albumin in a bottle. Maybe 100ml at a time, keep it
> refrigerated. When cutting nails, put 2-3 drops of albumin on the slide 
> and
> smear to cover all of the glass.
>
> We cut 3 sections of the nail specimen. 1 for H&E, 1 for PAS, and 1 extra
> for back-up. We bake the slides at 55'C overnight, and then at 70'C for 20
> minutes prior to staining the following day.
>
> This seems to work very well for us.
>
> ----- Original Message -----
> From: "Conlon, Paula F."
>
> To: "Histonet (E-mail)"
> Sent: Wednesday, June 14, 2006 9:16 AM
> Subject: [Histonet] toe nails
>
>
> Hi Histonetters, does anyone have a good procedure for sectioning nails 
> and
> keeping them on the slides during h&e staining and pas staining?Most of 
> the
> time our sections wash off. Thanks for your help.
>
>
> See our web page at http://www.lahey.org for a full directory of Lahey
> sites, staff, services and career opportunities.
>
> THIS MESSAGE IS INTENDED FOR THE USE OF THE PERSON TO WHOM IT IS 
> ADDRESSED.
> IT MAY CONTAIN INFORMATION THAT IS PRIVILEGED, CONFIDENTIAL AND EXEMPT 
> FROM
> DISCLOSURE UNDER APPLICABLE LAW. If you are not the intended recipient, 
> your
> use of this message for any purpose is strictly prohibited. If you have
> received this communication in error, please delete the message and notify
> the sender so that we may correct our records.
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
>
>
> ------------------------------
>
> Message: 9
> Date: Wed, 14 Jun 2006 08:52:10 -0700 (PDT)
> From: Geoffrey White
> Subject: [Histonet] Manual cover slip method - DiscoverSlip?
> To: histonet <@t> lists.utsouthwestern.edu
> Message-ID: <20060614155210.68428.qmail <@t> web36513.mail.mud.yahoo.com>
> Content-Type: text/plain; charset=iso-8859-1
>
>
> Dear all,
>
>
>
> at the time our research group is looking for new manual method for cover 
> our slides during the hybridization. In the past we used standard glass 
> cover slips but we are not really satisfied with this method.
>
> Just I saw at Biocompare a new method with the name DiscoverSlip. This 
> technology looks very interesting.
>
> Have any work with DiscoverSlip or saw this technology working?
>
>
>
> Thanks
>
> Geoffrey
>
> __________________________________________________
> Do You Yahoo!?
> Tired of spam? Yahoo! Mail has the best spam protection around
> http://mail.yahoo.com
>
> ------------------------------
>
> Message: 10
> Date: Wed, 14 Jun 2006 12:01:42 -0400
> From: Susan Q Wells
> Subject: Re: [Histonet] substrates
> To: Rene J Buesa
> Cc: histonet <@t> lists.utsouthwestern.edu, "Perry, Margaret"
>
> Message-ID: <449032E6.8030300 <@t> bms.com>
> Content-Type: text/plain; format=flowed; charset=ISO-8859-1
>
> You may lose your epitopes with this procedure, if so 10% H202 in PBS
> for 30 minutes may lighten the melanin pigment enough
> to view your chromagen. I recently used Vulcan Fast Red and Bajoran
> Purple from Biocare Medical with great success where melanin pigment was
> present.
>
> Sue Wells HT(ASCP), QIHC
>
> Rene J Buesa wrote:
>
>>Margaret:
>> Confronted with that situation I would eliminate the melanine better. 
>> Oxidize it with 0.1% potassium permanganate for 2 hours; wash and remove 
>> the oxidized melanine with 0.3% oxalic acid followed by washing. The 
>> section should be free of melanine pigment/colour afterwards and you can 
>> use DAB (for brown) and DAB + Ni salts for dark blue as double 
>> substrates.
>
> === message truncated ===
>
>
> ---------------------------------
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