[Histonet] mart-1 staining in Mohs
Mickley, Beth
Beth_Mickley <@t> URMC.Rochester.edu
Tue Feb 8 09:30:20 CST 2011
Hello,
Does anyone out there stain for mart-1 on their melanoma cases on frozen sections. I work for a Mohs surgeon in NYS and we are interested in doing our own testing in our lab for our melanoma cases. We are currently sending out our melanomas for slow Mohs. Any advice would be greatly appreciated.
Thank you!
________________________________________
From: histonet-bounces <@t> lists.utsouthwestern.edu [histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of histonet-request <@t> lists.utsouthwestern.edu [histonet-request <@t> lists.utsouthwestern.edu]
Sent: Monday, February 07, 2011 1:02 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: Histonet Digest, Vol 87, Issue 12
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Today's Topics:
1. RE: Amylase Digestion for glycogen (Tony Henwood)
2. pale stained areas H&E (raghulj <@t> orchidpharma.com)
3. BCL-1 staining on bone marrow core (emily stebens)
----------------------------------------------------------------------
Message: 1
Date: Mon, 7 Feb 2011 06:40:01 +0000
From: Tony Henwood <AnthonyH <@t> chw.edu.au>
Subject: RE: [Histonet] Amylase Digestion for glycogen
To: "'Ruppert, Amysue'" <ruppert.amysue <@t> marshfieldclinic.org>,
"histonet <@t> lists.utsouthwestern.edu"
<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <6D6BD1DE8A5571489398B392A38A715706250A <@t> xmdb02.nch.kids>
Content-Type: text/plain; charset="us-ascii"
Amysue,
Our amylase digestion procedure is quite simple (see: Mangan, V-M, Farago, V., Kelly, M., Henwood, A.F., (2002) "An Amylase Reagent with a Long Shelf Life for the removal of Glycogen from Tissue Sections" J. Histotechnol 25:153.) See below for the method:
Amylase Reagent
Warning: Harmful, contains azide - see MSDS
Alpha Amylase from Bacillus Subtilis (Fluka Cat No 10070,) 1g
Oxoid PBS Tablets (Cat No BR14a) 1 tablet
Distilled water 100ml
Sodium Azide 0.1g
This solution, once prepared is stored at 4oC when not in use. A recycled antibody dropper bottle (often used in commercial immunoperoxidase kits) is useful for storage and application.
Procedure:
1. Dewax and hydrate paraffin sections, hydrate frozen sections.
2. For amylase digestion, place slides on a rack, cover sections with amylase solution and allow to incubate for 10 minutes at room temperature.
3. Wash slides well in water.
4. Place slides in 1% periodic acid 10 minutes.
5. Wash slides well in water.
6. Rinse slides in distilled water.
7. Place in Schiff's reagent 10 minutes.
8. Rinse slides in distilled water and then wash slides in tap water 3 minutes.
9. Counterstain slides with haematoxylin, differentiate and blue.
10. Dehydrate, clear and mount.
Regards
Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA)
Laboratory Manager & Senior Scientist
Tel: 612 9845 3306
Fax: 612 9845 3318
the children's hospital at westmead
Cnr Hawkesbury Road and Hainsworth Street, Westmead
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Ruppert, Amysue
Sent: Saturday, 5 February 2011 11:03 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Amylase Digestion for glycogen
Hello,
We are looking to switch from malt diastase digestion for glycogen to Amylase digestion. I have the new protocol worked up, but one of the Pathologists I work with would like to have an idea of how many labs out there are using Amylase instead of malt diastase for their PAS/D method.
If you use amylase for the PAS/D method, could you please let me know who you are and the institiution?
Much appreciated.
amysue ruppert
Histology lab
Marshfield Labs
Marshfield WI
______________________________________________________________________
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------------------------------
Message: 2
Date: Mon, 7 Feb 2011 07:30:46 +0000
From: <raghulj <@t> orchidpharma.com>
Subject: [Histonet] pale stained areas H&E
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<A7F5262FC61D724E9AADD0FDBE8A327A17A2B732 <@t> EXCHMBX01.ORCHIDPHARMA.COM>
Content-Type: text/plain; charset="us-ascii"
Dear histonetters,
I have pasted two images img001 and img002 dt. 7feb2011 on staining. The images are of mouse kidney 4micron paraffin embedded formalin fixed sections where the hematoxylin stained areas are pale and not uniform. Is it because of over heating or sections getting dried during coverslipping?
Kindly help with your suggestions
Regards
raghul
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of histonet-request <@t> lists.utsouthwestern.edu
Sent: Saturday, February 05, 2011 11:32 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: Histonet Digest, Vol 87, Issue 10
Send Histonet mailing list submissions to
histonet <@t> lists.utsouthwestern.edu
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When replying, please edit your Subject line so it is more specific
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Today's Topics:
1. veterinary IHC for Ornithobacterium (Jan Shivers)
2. HCN4 staining (Shaw, Sharon)
3. Acetylcholinesterase (mtitford <@t> aol.com)
4. Artifact or dirt on slides using a tape cover slipper? (Nancy)
5. RE: Artifact or dirt on slides using a tape cover slipper?
(Setlak, Lisa)
6. RE: Artifact or dirt on slides using a tape cover slipper?
(Laurie Colbert)
7. Re: Karnovsky and Roots stain (John Kiernan)
8. RE: Karnovsky and Roots stain (Setlak, Lisa)
9. combined cholinesterase-silver stain (Nicole Cosenza)
10. pH Meter (Akemi Allison)
11. Amylase Digestion for glycogen (Ruppert, Amysue)
12. Re: Amylase Digestion for glycogen (John Kiernan)
13. Where can "Fast Blue" (Diamidino compound 253/50) be
obtained? (Per Magne Knutsen)
14. Re: Where can "Fast Blue" (Diamidino compound 253/50) be
obtained? (koellingr <@t> comcast.net)
15. Re: Amylase Digestion for glycogen (Rene J Buesa)
16. RE: SPAM-LOW: [Histonet] CE (Patsy Ruegg)
----------------------------------------------------------------------
Message: 1
Date: Fri, 4 Feb 2011 12:12:01 -0600
From: "Jan Shivers" <shive003 <@t> umn.edu>
Subject: [Histonet] veterinary IHC for Ornithobacterium
To: "histonet" <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <40B81232AD4048BA848C9586ABAA9255 <@t> auxs.umn.edu>
Content-Type: text/plain; charset="iso-8859-1"
Does anyone do IHC staining for Ornithobacterium rhinotracheale (ORT), a respiratory pathogen in poultry? I'm trying to find a lab to do it, or a vendor source for antibody.
Much thanks in advance,
Jan Shivers
Senior Scientist
Histology/IHC/EM Section Head
Pathology Teaching Program
University of Minnesota
Veterinary Diagnostic Laboratory
1333 Gortner Ave.
St. Paul, MN 55108
612-624-7297
shive003 <@t> umn.edu
(Confidentiality Notice: This message, together with any attachments, is intended only for the use of the individual or entity to which it is addressed and may contain confidential or privileged information. If you think you have received this message in error, please advise the sender and then delete this message and any attachments immediately.)
------------------------------
Message: 2
Date: Fri, 04 Feb 2011 14:41:38 -0500
From: "Shaw, Sharon" <shshaw <@t> WPI.EDU>
Subject: [Histonet] HCN4 staining
To: "histonet <@t> lists.utsouthwestern.edu"
<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<223F1D19A67B3245ADED1F18858077E005077552 <@t> S281.admin.wpi.edu>
Content-Type: text/plain; charset="us-ascii"
Hello,
I'm hoping somebody out there can give me some help with HCN4 staining, I was asked to stain HCN4 on mouse heart and it needs to stain the SA node. It is very hard to find that area since the node is very small. I have tried serial sections but not sure I hit the area or if I'm having problems with the stain.
The antibody is Rat monoclonal
Secondary Alexa Flour 488 goat anti-rat
Counter stain is Hoechst
I tried frozen and paraffin sections with no luck
Any suggestions will help greatly
Thanks,
Sharon
------------------------------
Message: 3
Date: Fri, 04 Feb 2011 15:06:18 -0500
From: mtitford <@t> aol.com
Subject: [Histonet] Acetylcholinesterase
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <8CD92BE1E6196A0-1C1C-38BD7 <@t> Webmail-d105.sysops.aol.com>
Content-Type: text/plain; charset="us-ascii"
Nicole Cosenza asks about acetylcholinesterase methods.
Years and years ago in London we used Koelle's method to demonstrate cholinesterase in muscle and mouse diaphragms. The method we followed was in Pearse E, Histochemistry - Theoretical and Applied. Volume 2. Churchill Livingstone. London 1972. Pages 1312 - 1316 has several different methods. The enzyme was visualized with ammonium sulphide. The tissues were mounted in glycerine jelly. If the method worked too well and we could not see the motor end plates, we adjusted the pH to reduce staining.
Dr Filipe went on to publish her own method in Filipe I., Lake B., Histochemistry in Pathology. Churchill Livingstone. London 1983 page 322. In her method, she used osmium to visualise the enzyme, instead of ammonium sulphide.
Stain technology used to have articles about the method too. I have not heard of Karnovsky or Roots methods, but lot of different methods were published in the early days of enzyme histochemistry.
Hope this helps
Michael Titford
Pathology - USA
Mobile AL
------------------------------
Message: 4
Date: Fri, 4 Feb 2011 12:12:13 -0800
From: "Nancy" <nancy <@t> pathologyarts.com>
Subject: [Histonet] Artifact or dirt on slides using a tape cover
slipper?
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <003a01cbc4a7$d0054560$700fd020$@com>
Content-Type: text/plain; charset="us-ascii"
At our facility with have 2 Tissue-Tek automated tape cover slippers. The
brand of tape that we use is made by a company called Klinipath, KP Tape
(dist. By Mercedes Medical).
On occasion we get complaints that the slides appear to have areas of dirt
or dust on them. It appears to be on the inside.
Has anyone else that uses a tape cover slipper ran across this particular
problem? If so, what did you do to troubleshoot? If it is the tape, is
there another brand or type that is preferable?
Nancy Mitchell
Pathology Arts, Inc
Director of Sales and Marketing
951-270-0605
909-732-1666-Cell
nancy <@t> pathologyarts.com
------------------------------
Message: 5
Date: Fri, 4 Feb 2011 14:22:50 -0600
From: "Setlak, Lisa" <LSetlak <@t> childrensmemorial.org>
Subject: RE: [Histonet] Artifact or dirt on slides using a tape cover
slipper?
To: 'Nancy' <nancy <@t> pathologyarts.com>,
"histonet <@t> lists.utsouthwestern.edu"
<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<7111DB39D045004C9CF29E79C71B28BC0CFA7683A3 <@t> CMHEXCC01MBX.childrensmemorial.org>
Content-Type: text/plain; charset="us-ascii"
We too have the same Coverslipper but we use the Sakura brand tape. We tried the KP tape but did not like it; it seemed to have problems sticking on the slides and we had a few other issues but I don't remember what they were. Could it be that there needs to be more xylene applied to slides- it may be air and dry tissue that you're seeing. Hope this helps.
Lisa M. Van Valkenberg, B.S., HT- ASCP
Histology Manager
2300 Children's Plaza
Chicago, IL 60614
773-868-8949
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Nancy
Sent: Friday, February 04, 2011 2:12 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Artifact or dirt on slides using a tape cover slipper?
At our facility with have 2 Tissue-Tek automated tape cover slippers. The
brand of tape that we use is made by a company called Klinipath, KP Tape
(dist. By Mercedes Medical).
On occasion we get complaints that the slides appear to have areas of dirt
or dust on them. It appears to be on the inside.
Has anyone else that uses a tape cover slipper ran across this particular
problem? If so, what did you do to troubleshoot? If it is the tape, is
there another brand or type that is preferable?
Nancy Mitchell
Pathology Arts, Inc
Director of Sales and Marketing
951-270-0605
909-732-1666-Cell
nancy <@t> pathologyarts.com
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 6
Date: Fri, 4 Feb 2011 12:44:19 -0800
From: "Laurie Colbert" <laurie.colbert <@t> huntingtonhospital.com>
Subject: RE: [Histonet] Artifact or dirt on slides using a tape cover
slipper?
To: "Nancy" <nancy <@t> pathologyarts.com>,
<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<57BE698966D5C54EAE8612E8941D76830A699BCD <@t> EXCHANGE3.huntingtonhospital.com>
Content-Type: text/plain; charset="us-ascii"
Nancy,
We get areas that appear brown when the tape doesn't lay flat or adhere
to the slide. Most of the time this happens on decal slides or hard
tissue where the tissue may be lifted a little. Try recoverslipping.
Also, if it's not happening on these types of tissue, you may need more
xylene dispensed. Sometimes, we just have to resort to the
old-fashioned way of coverslipping with mounting media. I've never used
the Klinipath tape. I prefer to stick with Sakura's brand - don't want
to take any chances!
Laurie Colbert
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Nancy
Sent: Friday, February 04, 2011 12:12 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Artifact or dirt on slides using a tape cover
slipper?
At our facility with have 2 Tissue-Tek automated tape cover slippers.
The
brand of tape that we use is made by a company called Klinipath, KP
Tape
(dist. By Mercedes Medical).
On occasion we get complaints that the slides appear to have areas of
dirt
or dust on them. It appears to be on the inside.
Has anyone else that uses a tape cover slipper ran across this
particular
problem? If so, what did you do to troubleshoot? If it is the tape, is
there another brand or type that is preferable?
Nancy Mitchell
Pathology Arts, Inc
Director of Sales and Marketing
951-270-0605
909-732-1666-Cell
nancy <@t> pathologyarts.com
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 7
Date: Fri, 04 Feb 2011 16:15:03 -0500
From: John Kiernan <jkiernan <@t> uwo.ca>
Subject: Re: [Histonet] Karnovsky and Roots stain
To: Nicole Cosenza <ncosenza <@t> siumed.edu>
Cc: "histonet <@t> lists.utsouthwestern.edu"
<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <fbdb95ab81a.4d4c2607 <@t> uwo.ca>
Content-Type: text/plain; CHARSET=US-ASCII
Karnovsky & Roots is (IMHO) the best histochemical method for choline esterase activity. In muscles, acetylcholinesterase (AChE) is the only such esterase shown by this method, and it is in the subneural apparatus of the motor endplate. Some counterstains (notably silver methods for the innervating axons) can remove the brown copper ferrocyanide product.
Another way to show motor endplates is with a method that picks up all esterases. In muscle, the endplate AChE shows up sooner than the enzymes present in all cells. Indigogenic esterase methods can be followed by silver staining of axons.
Why do you need or want to use fresh frozen, unfixed tissue sections? This makes no sense in the world of esterase activity histochemistry. There haven't been any developments in this field since the 1960s other than labelled alpha-bungarotoxin and immunohistochemistry.
An inexpensive book is Van Noorden CJF & Frederiks WM 1992. Enzyme Histochemistry. Oxford Univ Press and Royal Microscopical Soc. ISBN0198564341.
Another one is Lojda, Gossrau & Schiebler 1979. Enzyme histochemistry. Berlin: Springer. ISBN 0387092692.
A quick web search indicates that both are available and cost less than $10 second-hand.
John Kiernan
Anatomy, UWO
London, Canada
= = =
----- Original Message -----
From: Nicole Cosenza <ncosenza <@t> siumed.edu>
Date: Thursday, February 3, 2011 18:45
Subject: [Histonet] Karnovsky and Roots stain
To: histonet <@t> lists.utsouthwestern.edu
> I am looking into a project involving motor end plate staining.
> Literature that I've found continually references Karnovsky and
> Roots from the 60s. However the papers are not
> supplying all the details.
>
> Does anyone do AchE staining by this method on fresh frozen,
> unfixed tissue sections? If so, can I get a more detailed
> protocol (fixation steps, washes, etc)?
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 8
Date: Fri, 4 Feb 2011 15:20:19 -0600
From: "Setlak, Lisa" <LSetlak <@t> childrensmemorial.org>
Subject: RE: [Histonet] Karnovsky and Roots stain
To: 'John Kiernan' <jkiernan <@t> uwo.ca>, Nicole Cosenza
<ncosenza <@t> siumed.edu>
Cc: "histonet <@t> lists.utsouthwestern.edu"
<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<7111DB39D045004C9CF29E79C71B28BC0CFA7683A7 <@t> CMHEXCC01MBX.childrensmemorial.org>
Content-Type: text/plain; charset="us-ascii"
We do ACH staining on rectal biopsies to evaluate for ganglion cells for Hirschsprung's disease. Our stain is performed on fresh frozen tissue and I believe it's the Karnovsky method, but I'm not sure. Feel free to email if you are interested in out procedure.
Lisa M. Van Valkenberg, B.S., HT- ASCP
Histology Manager
2300 Children's Plaza
Chicago, IL 60614
773-868-8949
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of John Kiernan
Sent: Friday, February 04, 2011 3:15 PM
To: Nicole Cosenza
Cc: histonet <@t> lists.utsouthwestern.edu
Subject: Re: [Histonet] Karnovsky and Roots stain
Karnovsky & Roots is (IMHO) the best histochemical method for choline esterase activity. In muscles, acetylcholinesterase (AChE) is the only such esterase shown by this method, and it is in the subneural apparatus of the motor endplate. Some counterstains (notably silver methods for the innervating axons) can remove the brown copper ferrocyanide product.
Another way to show motor endplates is with a method that picks up all esterases. In muscle, the endplate AChE shows up sooner than the enzymes present in all cells. Indigogenic esterase methods can be followed by silver staining of axons.
Why do you need or want to use fresh frozen, unfixed tissue sections? This makes no sense in the world of esterase activity histochemistry. There haven't been any developments in this field since the 1960s other than labelled alpha-bungarotoxin and immunohistochemistry.
An inexpensive book is Van Noorden CJF & Frederiks WM 1992. Enzyme Histochemistry. Oxford Univ Press and Royal Microscopical Soc. ISBN0198564341.
Another one is Lojda, Gossrau & Schiebler 1979. Enzyme histochemistry. Berlin: Springer. ISBN 0387092692.
A quick web search indicates that both are available and cost less than $10 second-hand.
John Kiernan
Anatomy, UWO
London, Canada
= = =
----- Original Message -----
From: Nicole Cosenza <ncosenza <@t> siumed.edu>
Date: Thursday, February 3, 2011 18:45
Subject: [Histonet] Karnovsky and Roots stain
To: histonet <@t> lists.utsouthwestern.edu
> I am looking into a project involving motor end plate staining.
> Literature that I've found continually references Karnovsky and
> Roots from the 60s. However the papers are not
> supplying all the details.
>
> Does anyone do AchE staining by this method on fresh frozen,
> unfixed tissue sections? If so, can I get a more detailed
> protocol (fixation steps, washes, etc)?
>
> _______________________________________________
> 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: Fri, 04 Feb 2011 15:53:49 -0600
From: Nicole Cosenza <ncosenza <@t> siumed.edu>
Subject: [Histonet] combined cholinesterase-silver stain
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <4D4C756D.4000404 <@t> siumed.edu>
Content-Type: text/plain; charset=ISO-8859-1; format=flowed
I am looking into staining motor end plates. I've come across this
combined cholinesterase-silver stain (reference Pestronk and Drachman,
1978). Based on the date of the paper, I'm wondering what the current
technique is for this double staining.
Anyone currently doing AchE and axon staining on fresh frozen muscle
sections?
------------------------------
Message: 10
Date: Fri, 4 Feb 2011 14:04:56 -0800 (PST)
From: Akemi Allison <akemiat3377 <@t> yahoo.com>
Subject: [Histonet] pH Meter
To: histonet <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <506187.89374.qm <@t> web113811.mail.gq1.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1
Happy Friday everyone!
I am in the market for a new pH Meter, and was wondering if any of you had any
preferences.? I want to keep it simple as possible for?the staff.? Also, I
?don't want to spend a ton of money.
One of our fellow histonet subscribers?recommended purchasing a pH Meter that
did a? 2-step calibration verses a 3-step.? I've always used a 3-step
calibration.? Any thoughts.?
?
I realize that it must have a good glass electrode, calibrated daily, prior to
use, and all the maintenence must be followed to keep it in good working order.
?
If you have any suggestions, please provide the make and model #, and if you
have a vendor who provides it such as Fisher or Thermo, that would be an added
bonus.
Thank you, and have a great weekend!
Akemi Allison BS, HT(ASCP)HTL
Director
Phoenix Lab Consulting
E-Mail: akemiat3377 <@t> yahoo.com
------------------------------
Message: 11
Date: Fri, 04 Feb 2011 18:03:05 -0600
From: "Ruppert, Amysue" <ruppert.amysue <@t> marshfieldclinic.org>
Subject: [Histonet] Amylase Digestion for glycogen
To: "histonet <@t> lists.utsouthwestern.edu"
<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <201102050003.p15030nh007461 <@t> spamfilt>
Content-Type: text/plain; charset="iso-8859-1"
Hello,
We are looking to switch from malt diastase digestion for glycogen to Amylase digestion. I have the new protocol worked up, but one of the Pathologists I work with would like to have an idea of how many labs out there are using Amylase instead of malt diastase for their PAS/D method.
If you use amylase for the PAS/D method, could you please let me know who you are and the institiution?
Much appreciated.
amysue ruppert
Histology lab
Marshfield Labs
Marshfield WI
______________________________________________________________________
The contents of this message may contain private, protected and/or privileged information. If you received this message in error, you should destroy the e-mail message and any attachments or copies, and you are prohibited from retaining, distributing, disclosing or using any information contained within. Please contact the sender and advise of the erroneous delivery by return e-mail or telephone. Thank you for your cooperation.
------------------------------
Message: 12
Date: Sat, 05 Feb 2011 00:11:35 -0500
From: John Kiernan <jkiernan <@t> uwo.ca>
Subject: Re: [Histonet] Amylase Digestion for glycogen
To: "Ruppert, Amysue" <ruppert.amysue <@t> marshfieldclinic.org>,
"histonet <@t> lists.utsouthwestern.edu"
<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <7410bdbf6866.4d4c95b7 <@t> uwo.ca>
Content-Type: text/plain; charset="iso-8859-1"
<div style="font-family: 'times new roman'; font-size:
1 takes an did he lear ><br/></div><div>W the biology teacher told us that dias names for the enzyme in saliva and pancr starch.?This was also true in?my two prec medical school (early 1960s). The biochemistry teache more about starch and its animal equivalent glycogen, both digestible by amylase. Our early-1960s textbooks told us about
Claude properties demonstrated the?func homeostasis.</div><d ><br/></div><div>According to the Sigma catalogue, di astase is now "an obsolete synonym for
alpha-amylase". from many sources, includin beta-amylases, which would also catalyze glycogen.)? Sigma's?least expensive alpha-amyl human saliva.</div><div ><br/></div><div amylase will do the job. Go with the cheapest.?Human drooli are free and do not contain enzymes that will digest and solubilize polysaaccharides other than glygogen and starch. This is traditional
h papers f ><br/></div><div>Joh Kiernan</div><div>Anatomy, UWO</div><div>London,
C class= </b><ruppe wrote:</div><blockq cite="mid:201102050003.p15030nh007461 <@t> spamfilt" class= "iwcQuote" style="border-left: #00f 1px solid;
padding-lef type="cite"><div class=" plain">Hello,<br />?We are looking to switch f diastase digestion for glycogen to Amylase digestion. I have new protocol worked up, but one of the Pathologists I work with wo uld like to have an idea of how many labs out there are using Amylase
in you use who you are appreciated.<br /><br lab<br />Marshfield Labs<br />____________ _______________________ 5F _______________________ 5F message may information.? If you destroy the e-mail message are prohibited from retaining, any information contained within. advise of the erroneous delivery by re Thank you for your cooperation.<br /> </div></blockquote></div>
------------------------------
Message: 13
Date: Fri, 4 Feb 2011 23:22:41 -0800
From: Per Magne Knutsen <pknutsen <@t> physics.ucsd.edu>
Subject: [Histonet] Where can "Fast Blue" (Diamidino compound 253/50)
be obtained?
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
<AANLkTimXPaxovD+8_P3ZPQ1P7u6y2g1jGwtXWxodriPN <@t> mail.gmail.com>
Content-Type: text/plain; charset=UTF-8
Hi,
I've searched every catalog and website for "Fast Blue" (Diamidino compound
253/50), widely used for neuronal retrograde tracing. One recent paper using
this compound is:
Porreroa et al "Mapping of fluorescent protein-expressing neurons and axon
pathways in adult and developing Thy1-eYFP-H transgenic mice", Brain
Research, 1345, 59-72
where the authors as many before them cite "Dr. Illing GmbH & Co. KG,
Gro??-Umstadt, Germany" as the origin. I have been unable to locate this
particular vendor. I've written the authors and am waiting for an answer.
In the meanwhile, can anyone on this list help?
Kindly,
*Per M Knutsen
*Department of Physics
University of California, San Diego
9500 Gilman Drive, La Jolla, CA 92093-0374
T: +1 858 405 2868
E: pknutsen <@t> ucsd.edu
W: http://pmknutsen.blogspot.com
*
*
------------------------------
Message: 14
Date: Sat, 5 Feb 2011 15:20:01 +0000 (UTC)
From: koellingr <@t> comcast.net
Subject: Re: [Histonet] Where can "Fast Blue" (Diamidino compound
253/50) be obtained?
To: Per Magne Knutsen <pknutsen <@t> physics.ucsd.edu>
Cc: histonet <@t> lists.utsouthwestern.edu
Message-ID:
<666683375.5867.1296919201956.JavaMail.root <@t> sz0001a.emeryville.ca.mail.comcast.net>
Content-Type: text/plain; charset=utf-8
Hello Per,
Have no particular interest in neuronal retrograde tracing and have certainly never used anything for that. Just general scientific curiosity and a few minutes time awaiting a home project.?? Found Fast Blue (synonym: Diamidino compound 253/50) on Sigma website cat # F5756. Good luck.
Ray
Ray Koelling
PhenoPath Labs
Seattle WA
----- Original Message -----
From: "Per Magne Knutsen" <pknutsen <@t> physics.ucsd.edu>
To: histonet <@t> lists.utsouthwestern.edu
Sent: Friday, February 4, 2011 11:22:41 PM
Subject: [Histonet] Where can "Fast Blue" (Diamidino compound 253/50) be????????????????obtained?
Hi,
I've searched every catalog and website for "Fast Blue" (Diamidino compound
253/50), widely used for neuronal retrograde tracing. One recent paper using
this compound is:
Porreroa et al "Mapping of fluorescent protein-expressing neurons and axon
pathways in adult and developing Thy1-eYFP-H transgenic mice", Brain
Research, 1345, 59-72
where the authors as many before them cite "Dr. Illing GmbH & Co. KG,
Gro??-Umstadt, Germany" as the origin. I have been unable to locate this
particular vendor. I've written the authors and am waiting for an answer.
In the meanwhile, can anyone on this list help?
Kindly,
*Per M Knutsen
*Department of Physics
University of California, San Diego
9500 Gilman Drive, La Jolla, CA 92093-0374
T: +1 858 405 2868
E: pknutsen <@t> ucsd.edu
W: http://pmknutsen.blogspot.com
*
*
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------------------------------
Message: 15
Date: Sat, 5 Feb 2011 08:42:57 -0800 (PST)
From: Rene J Buesa <rjbuesa <@t> yahoo.com>
Subject: Re: [Histonet] Amylase Digestion for glycogen
To: "histonet <@t> lists.utsouthwestern.edu"
<histonet <@t> lists.utsouthwestern.edu>, AmysueRuppert
<ruppert.amysue <@t> marshfieldclinic.org>
Message-ID: <228486.12081.qm <@t> web65712.mail.ac4.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1
You are dealing with synonyms of the same enzyme but the "amylase" protocol always is slightly more complex.
Stick with "diastase" and, please, never use your own saliva for this procedure!!!!
Ren? J.
--- On Fri, 2/4/11, Ruppert, Amysue <ruppert.amysue <@t> marshfieldclinic.org> wrote:
From: Ruppert, Amysue <ruppert.amysue <@t> marshfieldclinic.org>
Subject: [Histonet] Amylase Digestion for glycogen
To: "histonet <@t> lists.utsouthwestern.edu" <histonet <@t> lists.utsouthwestern.edu>
Date: Friday, February 4, 2011, 7:03 PM
Hello,
We are looking to switch from malt diastase digestion for glycogen to Amylase digestion. I have the new protocol worked up, but one of the Pathologists I work with would like to have an idea of how many labs out there are using Amylase instead of malt diastase for their PAS/D method.
If you use amylase for the PAS/D method, could you please let me know who you are and the institiution?
Much appreciated.
amysue ruppert
Histology lab
Marshfield Labs
Marshfield WI
______________________________________________________________________
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Message: 16
Date: Sat, 5 Feb 2011 10:25:21 -0700
From: "Patsy Ruegg" <pruegg <@t> ihctech.net>
Subject: RE: SPAM-LOW: [Histonet] CE
To: "'andrea conard'" <andrea.conard <@t> gmail.com>,
<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <7B33835B42264FAE93FD4F474B9283CE <@t> prueggihctechlt>
Content-Type: text/plain; charset="US-ASCII"
It was ASCP that had TechSample, they do not have that anymore. The
teleconferences from NSH are reasonable and can be done at a distance.
Patsy Ruegg, HT(ASCP)QIHC
IHCtech
12635 Montview Blvd. Ste.215
Aurora, CO 80045
720-859-4060
fax 720-859-4110
www.ihctech.net
www.ihcrg.org
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of andrea
conard
Sent: Friday, February 04, 2011 10:18 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: SPAM-LOW: [Histonet] CE
I'm looking for some cost effective CE for my staff. Does anyone know if the
CAP still has TechSamples?
Please email at andrea.conard <@t> atlanticare.org
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End of Histonet Digest, Vol 87, Issue 10
****************************************
------------------------------
Message: 3
Date: Mon, 7 Feb 2011 10:44:48 -0500
From: emily stebens <emilystebens <@t> gmail.com>
Subject: [Histonet] BCL-1 staining on bone marrow core
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
<AANLkTi=dndQq=hFHWeA0HxUEJUDb5UxgXucjirw0AXhj <@t> mail.gmail.com>
Content-Type: text/plain; charset=ISO-8859-1
Does anyone have a protocol or tips for getting bcl-1 to work on
commercially decaled bone marrow?
------------------------------
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End of Histonet Digest, Vol 87, Issue 12
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