[Histonet] Re: Histonet Digest, Vol 98, Issue 6
Madeleine Huey
madeleinehuey <@t> gmail.com
Fri Jan 6 00:12:40 CST 2012
Sarah,
Your background is caused by cross-activity from your mouse primary
antibody on mouse tissue (xenografts). You can try Mouse on Mouse kit
(Biocare's has a great Mouse on Mouse HRP Polymer system).
I have used Cell Signaling's Rabbit anti-Cleaved Caspase 3 (Cat #
9661) on Xenografts and they work very well. They are very specified
and high affinity (appx. ~ 1:10k) with Dako's Envision + system.
Your problem will be solve if you just switch your primary ab from
mouse to rabbit.
Good Luck!
Madeleine Huey BS, HTL (ASCP) QIHC
Supervisor - Pathology (IPOX & Histology)
madeleinehuey <@t> elcaminohospital.org
On Thu, Jan 5, 2012 at 10:00 AM,
<histonet-request <@t> lists.utsouthwestern.edu> wrote:
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> Today's Topics:
>
> 1. RE: decal solution for molecular studies (Clare Thornton)
> 2. Amazing (Sarah Dysart)
> 3. RE: Amazing (Bartlett, Jeanine (CDC/OID/NCEZID))
> 4. Re: Amazing (Rene J Buesa)
> 5. RE: Amazing (Bernice Frederick)
> 6. RE: Amazing (Burton, Lynn)
> 7. Look at me... (Sarah Dysart)
> 8. Nail Polish sealant (gayle callis)
> 9. RE: Look at me... (Elizabeth Chlipala)
> 10. Elastichrome paper, and troubleshooting (Morken, Timothy)
> 11. RE: Elastichrome paper, and troubleshooting (Elizabeth Chlipala)
>
>
> ----------------------------------------------------------------------
>
> Message: 1
> Date: Thu, 5 Jan 2012 10:16:06 -0500
> From: Clare Thornton <CThornton <@t> dahlchase.com>
> Subject: [Histonet] RE: decal solution for molecular studies
> To: "'histonet <@t> lists.utsouthwestern.edu'"
> <histonet <@t> lists.utsouthwestern.edu>
> Message-ID:
> <C9D78FFC9D668B4CBEA4405F84697504F819B6F15A <@t> iris.dahlchase.net>
> Content-Type: text/plain; charset="us-ascii"
>
> To clarify, the Formical we use is a formic acid/EDTA solution.
>
>
> Clare J. Thornton, HTL(ASCP), QIHC
> Assistant Histology Supervisor
> Dahl-Chase Diagnostic Services
> 417 State Street, Suite 540
> Bangor, ME 04401
> cthornton <@t> dahlchase.com
>
>
>
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Clare Thornton
> Sent: Thursday, January 05, 2012 9:12 AM
> To: 'histonet <@t> lists.utsouthwestern.edu'
> Subject: [Histonet] decal solution for molecular studies
>
> Does anyone know of a decal solution that enables molecular studies (Her 2 FISH, EGFR by PCR) to be performed later? We use Formical for our decal solution. Her 2 FISH works about 50% of the time, EGFR almost never. We are looking at having to cut undecalcified bone today for both these studies, and we are not equipped to cut undecalcified bone. Any suggestions?
>
>
>
> Clare J. Thornton, HTL(ASCP), QIHC
> Assistant Histology Supervisor
> Dahl-Chase Diagnostic Services
> 417 State Street, Suite 540
> Bangor, ME 04401
> cthornton <@t> dahlchase.com
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
> ------------------------------
>
> Message: 2
> Date: Thu, 5 Jan 2012 15:59:35 +0000
> From: Sarah Dysart <sdysart <@t> mirnarx.com>
> Subject: [Histonet] Amazing
> To: "histonet <@t> lists.utsouthwestern.edu"
> <histonet <@t> lists.utsouthwestern.edu>
> Message-ID:
> <8A70A9B2ECDD084DACFE6C59FCF86D509C9766 <@t> SN2PRD0702MB098.namprd07.prod.outlook.com>
>
> Content-Type: text/plain; charset="us-ascii"
>
> I find it amazing sometimes when you don't do something for awhile how quickly your brain throws the information away. That being said...I know back in the day when I was learning histology we used to make our own acid alcohol solution (now where I am had a butt load of Clearifier so I was using that up). I don't want to buy that stuff anymore as making the solution is way cheaper and works just as well. I want to say it was like a 1% acid solution in alcohol?? What was the acid? For some reason my brain says glacial acetic...but time has made me forget. Is the alcohol you mix it in 100% or something lower with a water content to it? Please help my alzheimers =)
> Happy Thursday!!
>
> Sarah Goebel-Dysart, BA, HT(ASCP)
> Histotechnologist
> Mirna Therapeutics
> 2150 Woodward Street
> Suite 100
> Austin, Texas 78744
> (512)901-0900 ext. 6912
>
>
>
> ------------------------------
>
> Message: 3
> Date: Thu, 5 Jan 2012 16:08:36 +0000
> From: "Bartlett, Jeanine (CDC/OID/NCEZID)" <jqb7 <@t> cdc.gov>
> Subject: [Histonet] RE: Amazing
> To: Sarah Dysart <sdysart <@t> mirnarx.com>,
> "histonet <@t> lists.utsouthwestern.edu"
> <histonet <@t> lists.utsouthwestern.edu>
> Message-ID:
> <DF1CBA3D83D9A344A7D6A045188E4484207A1C84 <@t> EMBX-CLFT1.cdc.gov>
> Content-Type: text/plain; charset="us-ascii"
>
> If you are talking about a regressive H&E stain then perhaps it was 1% HCL acid in 70% alcohol
>
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Sarah Dysart
> Sent: Thursday, January 05, 2012 11:00 AM
> To: histonet <@t> lists.utsouthwestern.edu
> Subject: [Histonet] Amazing
>
> I find it amazing sometimes when you don't do something for awhile how quickly your brain throws the information away. That being said...I know back in the day when I was learning histology we used to make our own acid alcohol solution (now where I am had a butt load of Clearifier so I was using that up). I don't want to buy that stuff anymore as making the solution is way cheaper and works just as well. I want to say it was like a 1% acid solution in alcohol?? What was the acid? For some reason my brain says glacial acetic...but time has made me forget. Is the alcohol you mix it in 100% or something lower with a water content to it? Please help my alzheimers =) Happy Thursday!!
>
> Sarah Goebel-Dysart, BA, HT(ASCP)
> Histotechnologist
> Mirna Therapeutics
> 2150 Woodward Street
> Suite 100
> Austin, Texas 78744
> (512)901-0900 ext. 6912
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
> ------------------------------
>
> Message: 4
> Date: Thu, 5 Jan 2012 08:16:48 -0800 (PST)
> From: Rene J Buesa <rjbuesa <@t> yahoo.com>
> Subject: Re: [Histonet] Amazing
> To: "histonet <@t> lists.utsouthwestern.edu"
> <histonet <@t> lists.utsouthwestern.edu>, Sarah Dysart
> <sdysart <@t> mirnarx.com>
> Message-ID:
> <1325780208.90675.YahooMailClassic <@t> web65703.mail.ac4.yahoo.com>
> Content-Type: text/plain; charset=iso-8859-1
>
> You can use either acetic or hydrochloric, both will do. The strength is 1% BUT it does not matter. As a matter of fact the weaker the solution the better. Remember that the acid solution (that can be made with 7ethanolanol even better) is used to differentiate progressivesive hematoxylin, like Harris, and the weaker it is the more control you have with the differentiation.
> A weak solution allows you to dip the slides several times until you obtain the desired reddish hue on the sections that signals when the differentiation is completed.
> If the solution is too strong you will have to take the slides often precipitouslyusly and some sections will be have a weak nuclear staining.
> So, use acetic at 1% in 70% ethanol That is what I used to prepare.
> René J.
>
> --- On Thu, 1/5/12, Sarah Dysart <sdysart <@t> mirnarx.com> wrote:
>
>
> From: Sarah Dysart <sdysart <@t> mirnarx.com>
> Subject: [Histonet] Amazing
> To: "histonet <@t> lists.utsouthwestern.edu" <histonet <@t> lists.utsouthwestern.edu>
> Date: Thursday, January 5, 2012, 10:59 AM
>
>
> I find it amazing sometimes when you don't do something for awhile how quickly your brain throws the information away. That being said...I know back in the day when I was learning histology we used to make our own acid alcohol solution (now where I am had a butt load of Clearifier so I was using that up). I don't want to buy that stuff anymore as making the solution is way cheaper and works just as well. I want to say it was like a 1% acid solution in alcohol?? What was the acid? For some reason my brain says glacial acetic...but time has made me forget. Is the alcohol you mix it in 100% or something lower with a water content to it? Please help my alzheimers =)
> Happy Thursday!!
>
> Sarah Goebel-Dysart, BA, HT(ASCP)
> Histotechnologist
> Mirna Therapeutics
> 2150 Woodward Street
> Suite 100
> Austin, Texas 78744
> (512)901-0900 ext. 6912
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
> ------------------------------
>
> Message: 5
> Date: Thu, 5 Jan 2012 16:24:51 +0000
> From: Bernice Frederick <b-frederick <@t> northwestern.edu>
> Subject: RE: [Histonet] Amazing
> To: Rene J Buesa <rjbuesa <@t> yahoo.com>,
> "histonet <@t> lists.utsouthwestern.edu"
> <histonet <@t> lists.utsouthwestern.edu>, Sarah Dysart
> <sdysart <@t> mirnarx.com>
> Message-ID:
> <62C639732D3F274DACED033EBDF6ADAF1E19DC42 <@t> evcspmbx3.ads.northwestern.edu>
>
> Content-Type: text/plain; charset="iso-8859-1"
>
> We use .5% acid alcohol ( hydrochloric) with Harris hemo. No problems.
> Bernice
>
> Bernice Frederick HTL (ASCP)
> Senior Research Tech
> Pathology Core Facility
> ECOGPCO-RL
> Robert. H. Lurie Cancer Center
> Northwestern University
> 710 N Fairbanks Court
> Olson 8-421
> Chicago,IL 60611
> 312-503-3723
> b-frederick <@t> northwestern.edu
>
>
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Rene J Buesa
> Sent: Thursday, January 05, 2012 10:17 AM
> To: histonet <@t> lists.utsouthwestern.edu; Sarah Dysart
> Subject: Re: [Histonet] Amazing
>
> You can use either acetic or hydrochloric, both will do. The strength is 1% BUT it does not matter. As a matter of fact the weaker the solution the better. Remember that the acid solution (that can be made with 7ethanolanol even better) is used to differentiate progressivesive hematoxylin, like Harris, and the weaker it is the more control you have with the differentiation.
> A weak solution allows you to dip the slides several times until you obtain the desired reddish hue on the sections that signals when the differentiation is completed.
> If the solution is too strong you will have to take the slides often precipitouslyusly and some sections will be have a weak nuclear staining.
> So, use acetic at 1% in 70% ethanol That is what I used to prepare.
> René J.
>
> --- On Thu, 1/5/12, Sarah Dysart <sdysart <@t> mirnarx.com> wrote:
>
>
> From: Sarah Dysart <sdysart <@t> mirnarx.com>
> Subject: [Histonet] Amazing
> To: "histonet <@t> lists.utsouthwestern.edu" <histonet <@t> lists.utsouthwestern.edu>
> Date: Thursday, January 5, 2012, 10:59 AM
>
>
> I find it amazing sometimes when you don't do something for awhile how quickly your brain throws the information away. That being said...I know back in the day when I was learning histology we used to make our own acid alcohol solution (now where I am had a butt load of Clearifier so I was using that up). I don't want to buy that stuff anymore as making the solution is way cheaper and works just as well. I want to say it was like a 1% acid solution in alcohol?? What was the acid? For some reason my brain says glacial acetic...but time has made me forget. Is the alcohol you mix it in 100% or something lower with a water content to it? Please help my alzheimers =) Happy Thursday!!
>
> Sarah Goebel-Dysart, BA, HT(ASCP)
> Histotechnologist
> Mirna Therapeutics
> 2150 Woodward Street
> Suite 100
> Austin, Texas 78744
> (512)901-0900 ext. 6912
>
> _______________________________________________
> 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: 6
> Date: Thu, 5 Jan 2012 10:25:27 -0600
> From: "Burton, Lynn" <Lynn.Burton <@t> Illinois.gov>
> Subject: [Histonet] RE: Amazing
> To: Sarah Dysart <sdysart <@t> mirnarx.com>,
> "histonet <@t> lists.utsouthwestern.edu"
> <histonet <@t> lists.utsouthwestern.edu>
> Message-ID:
> <4A6E2CACA1E017408EBA1B9911952CC006483C0722 <@t> IL084EXMBX214.illinois.gov>
>
> Content-Type: text/plain; charset="us-ascii"
>
> I still use that. It is 70% with 5mL of hydrochloric acid if making 1L.
> Lynn Burton
> Lab Assoc I
> Animal Disease Lab
> Galesburg, Il
> 309-344-2451
> ________________________________________
> From: histonet-bounces <@t> lists.utsouthwestern.edu [histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Sarah Dysart [sdysart <@t> mirnarx.com]
> Sent: Thursday, January 05, 2012 9:59 AM
> To: histonet <@t> lists.utsouthwestern.edu
> Subject: [Histonet] Amazing
>
> I find it amazing sometimes when you don't do something for awhile how quickly your brain throws the information away. That being said...I know back in the day when I was learning histology we used to make our own acid alcohol solution (now where I am had a butt load of Clearifier so I was using that up). I don't want to buy that stuff anymore as making the solution is way cheaper and works just as well. I want to say it was like a 1% acid solution in alcohol?? What was the acid? For some reason my brain says glacial acetic...but time has made me forget. Is the alcohol you mix it in 100% or something lower with a water content to it? Please help my alzheimers =)
> Happy Thursday!!
>
> Sarah Goebel-Dysart, BA, HT(ASCP)
> Histotechnologist
> Mirna Therapeutics
> 2150 Woodward Street
> Suite 100
> Austin, Texas 78744
> (512)901-0900 ext. 6912
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
> ------------------------------
>
> Message: 7
> Date: Thu, 5 Jan 2012 17:16:43 +0000
> From: Sarah Dysart <sdysart <@t> mirnarx.com>
> Subject: [Histonet] Look at me...
> To: "histonet <@t> lists.utsouthwestern.edu"
> <histonet <@t> lists.utsouthwestern.edu>
> Message-ID:
> <8A70A9B2ECDD084DACFE6C59FCF86D509C9831 <@t> SN2PRD0702MB098.namprd07.prod.outlook.com>
>
> Content-Type: text/plain; charset="us-ascii"
>
> I'm just full of questions today!! This one is IHC...I have been trying to optimize a Caspase3 stain for several months now and it is still just chalked full of background gradoo. I do all the blocking including Fc receptors...still junk. The clone I have been using is, from abcam (ab2302). I don't see the specific clone name listed. I am staining human xenografts raised in mouse. I get a whole lot of background staining making it very hard to find the positive staining. The recommended dilution is about 1:30, but I have diluted all the way up to 1:500. At the higher dilution no positive staining or background is observed. Does anyone know of a good Caspase3 antibody, preferably mouse monoclonal? All the rabbit polyclonal antibodies are difficult to stain on these xenografts.
> Thanks again =)
>
> Sarah Goebel-Dysart, BA, HT(ASCP)
> Histotechnologist
> Mirna Therapeutics
> 2150 Woodward Street
> Suite 100
> Austin, Texas 78744
> (512)901-0900 ext. 6912
>
>
>
> ------------------------------
>
> Message: 8
> Date: Thu, 5 Jan 2012 10:35:20 -0700
> From: "gayle callis" <gayle.callis <@t> bresnan.net>
> Subject: [Histonet] Nail Polish sealant
> To: <kasaim <@t> mail.nih.gov>, "Histonet"
> <histonet <@t> lists.utsouthwestern.edu>
> Message-ID: <000e01cccbd0$67966f50$36c34df0$@bresnan.net>
> Content-Type: text/plain; charset="us-ascii"
>
> You wrote:
>
> We are currently starting up some IHC on frozen tissue sections. After
> staining with different fluorescent antibodies, we end with applying DAPI
> w/Prolong gold and then coverslipping. We would like to seal the coverslip
> so that we can keep the slides longer. Any suggestions on where and how
> best to apply the nail polish for a permanent fix on the coverslips?
>
>
>
> *****************************
>
> Prolong Gold antifade reagent is a hard seal to begin with. We only seal
> ends but not the sides of cover glass. Our coverslips go right to edge of
> slides e.g. 25 X 30, 25 x 40, or 25 X 50. We don't like having nail
> polish slop over the edges onto back of slides but one could seal all sides
> of coverslip if careful. We buy the thinner top or base coat nail
> polish, but in general, prefer to use thinned mounting media rather than
> nail polish. There can be some issues here. If you are trying to view
> GFP or RFP (red fluorescence protein) labeled cells or tissue components,
> you should not seal the coverslip with nail polish since the alcohol in nail
> polish leaches under the coverslip and causes GFP/RFP to fade. This fact
> was published in Science. We found that dumping out cheap clear nail
> polish from bottle, rinsing away the residue with acetone, and then adding
> permanent mounting media and thinning that with toluene to the consistency
> of top coat nail polish works best. Toluene or xylene based sealants cannot
> leach under the cover glass since these solvents are NOT miscible with water
> in the PBS. Thinned mounting media is better sealant for GFP purposes (no
> fading) and also works for IF stained sections (perfect seal). We love the
> little brush in the nail polish bottle for application. Thicker clear nail
> polish (for non GFP studies) or IF stained sections is messy during
> application so we buy the cheapest top coat polish we can find at Walmart.
>
>
>
>
> DAPI in the Prolong Gold will cause an uneven staining gradient so that some
> of the nuclei in the center of a section are not stained as brightly as the
> nuclei on the outer edges of a stained section. The cause is not getting
> enough thicker Prolong Gold/DAPI over the section or not having just the
> right amount of buffer on the section to permit a good flow of this
> wonderful mounting media over the section. We now complete all IF
> staining then stain with a DAPI solution before cover slipping with Prolong
> Gold. You can buy ready to use DAPI solutions from Pierce or Biogenex, or
> make up the solution in house. You can find the recipe at IHCworld website
> or simply Google.
>
>
>
> We do NOT store our IF stained slides in the cold, but in a folder at RT in
> a dark drawer before viewing on the day after staining to reduce any
> movement/flow under the coverslip. Fluorophores can and will eventually
> fade. I do not recall any studies saying storing IF stained slides in the
> cold reduces fading but we never have space to do cold storage anyway and
> store slides at RT. The new fluorophores (Alexas and Dylights) remain
> stable over a longer time even for several weeks compared to fluorescein
> derivatives e.g. FITC TRITC.
>
>
>
> Gayle M. Callis
>
> HTL/HT/MT(ASCP)
>
> Bozeman MT
>
>
>
> ------------------------------
>
> Message: 9
> Date: Thu, 5 Jan 2012 10:36:16 -0700
> From: Elizabeth Chlipala <liz <@t> premierlab.com>
> Subject: [Histonet] RE: Look at me...
> To: 'Sarah Dysart' <sdysart <@t> mirnarx.com>,
> "histonet <@t> lists.utsouthwestern.edu"
> <histonet <@t> lists.utsouthwestern.edu>
> Message-ID:
> <14E2C6176416974295479C64A11CB9AE011380AC7447 <@t> SBS2K8.premierlab.local>
> Content-Type: text/plain; charset="us-ascii"
>
> If you are looking to stain cleaved caspase 3 there is a better antibody from cell signaling it works in multiple species, it's a bit pricey but I have found that it works the best out of the few that I have tried. We have used it mouse xenografts before without any issue.
>
> Liz
>
> Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
> Manager
> Premier Laboratory, LLC
> PO Box 18592
> Boulder, CO 80308-1592
> (303) 682-3949 office
> (303) 682-9060 fax
> (303) 881-0763 cell
> www.premierlab.com
>
> Ship to address:
>
> 1567 Skyway Drive, Unit E
> Longmont, CO 80504
>
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Sarah Dysart
> Sent: Thursday, January 05, 2012 10:17 AM
> To: histonet <@t> lists.utsouthwestern.edu
> Subject: [Histonet] Look at me...
>
> I'm just full of questions today!! This one is IHC...I have been trying to optimize a Caspase3 stain for several months now and it is still just chalked full of background gradoo. I do all the blocking including Fc receptors...still junk. The clone I have been using is, from abcam (ab2302). I don't see the specific clone name listed. I am staining human xenografts raised in mouse. I get a whole lot of background staining making it very hard to find the positive staining. The recommended dilution is about 1:30, but I have diluted all the way up to 1:500. At the higher dilution no positive staining or background is observed. Does anyone know of a good Caspase3 antibody, preferably mouse monoclonal? All the rabbit polyclonal antibodies are difficult to stain on these xenografts.
> Thanks again =)
>
> Sarah Goebel-Dysart, BA, HT(ASCP)
> Histotechnologist
> Mirna Therapeutics
> 2150 Woodward Street
> Suite 100
> Austin, Texas 78744
> (512)901-0900 ext. 6912
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
> ------------------------------
>
> Message: 10
> Date: Thu, 5 Jan 2012 09:45:25 -0800
> From: "Morken, Timothy" <Timothy.Morken <@t> ucsfmedctr.org>
> Subject: [Histonet] Elastichrome paper, and troubleshooting
> To: Histonet <histonet <@t> lists.utsouthwestern.edu>
> Message-ID:
> <8D7C2D242DBD45498006B21122072BF89F5EE62C <@t> MCINFRWEM003.ucsfmedicalcenter.org>
>
> Content-Type: text/plain; charset=us-ascii
>
> Does anyone have a pdf (or could fax me) the original paper for "Elastichrome" stain:
>
> Richardson, L. "Combination Elastic Trichrome Stain," Laboratory Medicine, 6.1, 1975
>
> We have a procedure with this reference, but no original paper to look at. We do it very rarely and no one seems to know how well it worked in the past.
>
> I do have another paper in the same vein, "A combination verhoeff's elastic and masson's trichrom stain for routine histology," O'Connor, S. Valle, Stain Technology V 57, no. 4, 1982, that is a modification of the Richardson paper, and gives us some ideas, but I would still like to see the original paper if possible.
>
> Our problem is that the elastic stain washes out during or after the trichrome.
>
> Procedure:
> Bouins, 56C one hour
> Wiegerts hematolylin, 3min
> Verhoffs elastic stain, 15 min
> 2% ferric chloride differentiation (so far so-good)
> Gomori's trichrome Blue, 15 min
> 0.2% glacial acetic acid, 1 min
> Wash dH20, dehydrate, clear, mount
>
> Results: trichrome looks great, no elastin stain.
>
> I'm thinking the acetic acid is too long so told them to shorten it to a few dips and see how it looks. But, maybe some has some experience with this and has other suggestions.
>
> Thanks for any help.
>
>
> Tim Morken
> Supervisor, Histology, IPOX
> UC San Francisco Medical Center
> Box 1656
> 1600 Divisidero St, B217
> San Francisco, CA 94115
> USA
>
> 415.514.6042 (office)
> 415.885.7409 Fax
> tim.morken <@t> ucsfmedctr.org<mailto:tim.morken <@t> ucsfmedctr.org>
>
>
>
> ------------------------------
>
> Message: 11
> Date: Thu, 5 Jan 2012 10:50:05 -0700
> From: Elizabeth Chlipala <liz <@t> premierlab.com>
> Subject: [Histonet] RE: Elastichrome paper, and troubleshooting
> To: "'Morken, Timothy'" <Timothy.Morken <@t> ucsfmedctr.org>, Histonet
> <histonet <@t> lists.utsouthwestern.edu>
> Message-ID:
> <14E2C6176416974295479C64A11CB9AE011380AC7449 <@t> SBS2K8.premierlab.local>
> Content-Type: text/plain; charset="us-ascii"
>
> Tim
>
> We do this stain all of the time, we never used the original reference we just made up one on our own. I'll send our SOP in a different e-mail.
>
> Liz
>
> Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
> Manager
> Premier Laboratory, LLC
> PO Box 18592
> Boulder, CO 80308-1592
> (303) 682-3949 office
> (303) 682-9060 fax
> (303) 881-0763 cell
> www.premierlab.com
>
> Ship to address:
>
> 1567 Skyway Drive, Unit E
> Longmont, CO 80504
>
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Morken, Timothy
> Sent: Thursday, January 05, 2012 10:45 AM
> To: Histonet
> Subject: [Histonet] Elastichrome paper, and troubleshooting
>
> Does anyone have a pdf (or could fax me) the original paper for "Elastichrome" stain:
>
> Richardson, L. "Combination Elastic Trichrome Stain," Laboratory Medicine, 6.1, 1975
>
> We have a procedure with this reference, but no original paper to look at. We do it very rarely and no one seems to know how well it worked in the past.
>
> I do have another paper in the same vein, "A combination verhoeff's elastic and masson's trichrom stain for routine histology," O'Connor, S. Valle, Stain Technology V 57, no. 4, 1982, that is a modification of the Richardson paper, and gives us some ideas, but I would still like to see the original paper if possible.
>
> Our problem is that the elastic stain washes out during or after the trichrome.
>
> Procedure:
> Bouins, 56C one hour
> Wiegerts hematolylin, 3min
> Verhoffs elastic stain, 15 min
> 2% ferric chloride differentiation (so far so-good)
> Gomori's trichrome Blue, 15 min
> 0.2% glacial acetic acid, 1 min
> Wash dH20, dehydrate, clear, mount
>
> Results: trichrome looks great, no elastin stain.
>
> I'm thinking the acetic acid is too long so told them to shorten it to a few dips and see how it looks. But, maybe some has some experience with this and has other suggestions.
>
> Thanks for any help.
>
>
> Tim Morken
> Supervisor, Histology, IPOX
> UC San Francisco Medical Center
> Box 1656
> 1600 Divisidero St, B217
> San Francisco, CA 94115
> USA
>
> 415.514.6042 (office)
> 415.885.7409 Fax
> tim.morken <@t> ucsfmedctr.org<mailto:tim.morken <@t> ucsfmedctr.org>
>
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> End of Histonet Digest, Vol 98, Issue 6
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