[Histonet] RE: Acetylcholonesterase

From:"Driessen, L."

This recipe works well

1. Prepare the incubation-medium.
2. Incubate for 60-90 minutes at 37oC.
3. 3 min. Saturated NaSO4-solution.
4. 2 min. 1% ammoniumsulfideoplossing (fumehead!!!).
5. Rinse with tapwater.
6. 20 sec. Mayer-haematoxylin.
7. Rinse with tapwater for 5 min.
8. Mount in in glycerin-gelatin.


1. Cu-complexsolution:
(Dissolve in this order)
- 85 ml. Dist. water.
- 34 g. NaSO4.10 H2O.
- 150 mg. CuSO4.5H2O.
- 187 mg. glycin.
- 500 mg. MgCl2
- 875 mgr. maleinacid.
- Adjust pH to 5,6-6,0 with 15 ml. 1N NaOH.
	(Store in refrigerator; crystalyses when cold).

2. Incubation-medium:	Dissolve 10 mg. acetylthiocholinejodid in 5 ml. Cu-complexsolution.

3. 1% ammoniumsulfide: Ad 4 ml. ammoniumsulfide (20%) to 76 ml. dist. water.

Leon Driessen
ORL/UMC-Nijmegen, The Netherlands
l.driessen@orthop.umcn.nl


-----Original Message-----
From: histonet-request@lists.utsouthwestern.edu
[mailto:histonet-request@lists.utsouthwestern.edu]
Sent: woensdag 17 september 2003 19:00
To: histonet@lists.utsouthwestern.edu
Subject: Histonet digest, Vol 1 #51 - 30 msgs


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Today's Topics:

   1. Re: microwave (Sarah Jones)
   2. Acetylcholonesterase (WWmn916@aol.com)
   3. Re: Histonet digest, Vol 1 #50 - 11 msgs (amosbrooks@earthlink.net)
   4. Glass cleaning for silver (John Kiernan)
   5. RE: immunohistochemistry and frozen sections (C.M. van der Loos)
   6. acid cleaning slides for silver preps (George Cole)
   7. RE: S100 frozen skin (Mandy Townsend)
   8. Yarrow (peptolab)
   9. BrdU and EM? (Mezey Szilvia)
  10. job openings in Dallas (Priscilla Delventhal)
  11. KnifeMaker (Joyce Christopher)
  12. receiving un-registered specimens (Amy Self)
  13. RE: BrdU and EM? (Mezey Szilvia)
  14. wage/vacancy survey (Dawson, Glen)
  15. Used JB-4 Microtome? (DJStashev@aol.com)
  16. Re: KnifeMaker (Gayle Callis)
  17. Re: questions regarding to a-Naphthyl Butyrate
       Esterase stain and a-Naphthyl Acetate stain(no (Fred Underwood)
  18. Re: microwave (Claye Clyatt)
  19. Re: Acetylcholonesterase (Claye Clyatt)
  20. RE: questions regarding to a-Naphthyl ButyrateEsterase stain and a-Naphthyl Acetate stain(no (Pamela Marcum)
  21. RE: questions regarding to a-Naphthyl ButyrateEsterase stain and a-Naphthyl Acetate stain(no (Pamela Marcum)
  22. RE: questions regarding to a-Naphthyl ButyrateEsterase stain and a-Naphthyl Acetate stain(no (Pamela Marcum)
  23. Re: receiving un-registered specimens (Vicki Gauch)
  24. RE: receiving un-registered specimens (Morken, Tim - Labvision)
  25. RE: questions regarding to a-Naphthyl ButyrateEsterase stain and a-Naphthyl Acetate stain(no (Pamela Marcum)
  26. Ret IHC (Richard Cartun)
  27. Underprocessed tissue (Michelle D. Moore)
  28. Clot sections (Andrew Fedanov)
  29. Re: Ret IHC (DDittus787@aol.com)

--__--__--

Message: 1
Date: Tue, 16 Sep 2003 17:08:09 -0500
From: "Sarah Jones" 
To: ,
Subject: Re: [Histonet] microwave

Hello Gudrun,
   I'm glad to see you have posted a question.  I don't have a lot of
experience with IHC and the microwave, but I can tell you that the
microwave can shorten the times of many special stains.  Many steps that
usually take one hour can be shortened to just a few minutes.  Silver
stains work very well in the microwave.  There is a great book on
microwaving special stains called "Microwave Magic" by Billie Beck,
University of Texas Southwestern Medical Center in Dallas Texas.  It's
old, from 1987, so I don't know if it is still available.  Some of the
stains listed are, Grocott's Methenamine Silver, Jones Methenamine
Silver,  Fontana-Masson, Dieterle, Grimelius, Steiner and Steiner,
Seiver-Munger, Bielschowsky's, PAS, Mucicarmine, Alcian Blue, Colloidal
Iron, Rhodanine, Iron Stains, Trichrome, Acid Fast, Giemsa, Luxol Fast
Blue, Congo Red, PTAH, Movat's, Dunn Thompson, Bouins pre-treatment.  
  You can use a household microwave, but you have to know the wattage
of your oven.  Most of them range from 700-900 watts, but if it is lower
wattage, you would just leave it in longer.  Usually you would want to
determine how long it would take a coplin jar of water to reach 60
degrees C.  Also determine how long it would take different ethanol to
reach 60 degrees C.  You can also make a temperature conversion chart
that shows various solutions, times and power levels and what
temperature is reached. 
  Use plastic coplin jars, the glass ones will break, stir solutions
immediately as they come out of the oven (I use a wooden stick), and
cover the coplin jars with a gauze square when microwaving.  
  If you go back to using chemicals in the microwave, I wouldn't
continue to heat tea in it, just for safety reasons.  
  Happy Microwaving!   Sarah

Sarah Jones HT(ASCP)
Dept. of Vet. Anatomy & Public Health
Histology Lab
Texas A&M University
College Station, TX 77843-4458
phone: 979-845-3177
fax:  979-458-3499


>>> "Gudrun Lang"  09/16/03 04:14PM >>>
Hi Histonetters!
I have a question about microwave oven in histolabs. We did introduce
such a household thing for IHC, but it was not very successfull. Now we
heat our tea with it. IHC works with heat and steam.
Which equipment do you use for special staining? Are there specific
apparats or also the houshold ones? And has it become really common to
work with it?
thanks for your answers.

Gundi Lang
general hospital Linz, Austria


--__--__--

Message: 2
From: WWmn916@aol.com
Date: Tue, 16 Sep 2003 19:02:11 EDT
To: histonet@pathology.swmed.edu
Subject: [Histonet] Acetylcholonesterase


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Anyone have an easy recipe (or refer me to a prepared kit that can be 
purchased) for the acetylcholonesterase stain?  Doctor is looking for Hirschsprungs 
disease.  Any help is appreciated.

Sincerely,
Deb King, HT (ASCP)
Sacramento, CA

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Anyone have an easy recipe (or refer me to a prepared=20=
kit that can be purchased) for the acetylcholonesterase stain?  Doctor=20=
is looking for Hirschsprungs disease.  Any help is appreciated.

Sincerely,
Deb King, HT (ASCP)
Sacramento, CA
--part1_1d1.10ef4df6.2c98f073_boundary-- --__--__-- Message: 3 From: amosbrooks@earthlink.net To: histonet@lists.utsouthwestern.edu Date: Tue, 16 Sep 2003 22:19:36 -0400 Subject: [Histonet] Re: Histonet digest, Vol 1 #50 - 11 msgs Hi, We use a Black & Decker vegtable steamer. I bored a hole into the top of the steamer and we put a thermometer directly into the solution to get a Real Time measurement of the temperature. (BTW CAP loved it) We also use a waterbath as is indicated by the Herceptest (DAKO) instructions. I strongly dislike the use of household microwaves for any laboratory purposes, especially something as finiky as antigen retrieval. The temperature over time spiles up to an overheated state then drops rapidly to a point that it is not retrieving at all only to be reheated in the same way. It is impossible to have any uniformity this way. You're much better off with a steady heat source for a specific amount of time. Haste makes waste, Amos Brooks Hi Histonetters! I have a question about microwave oven in histolabs. We did introduce such a household thing for IHC, but it was not very successfull. Now we heat our tea with it. IHC works with heat and steam. Which equipment do you use for special staining? Are there specific apparats or also the houshold ones? And has it become really common to work with it? thanks for your answers. Gundi Lang general hospital Linz, Austria --__--__-- Message: 4 Date: Wed, 17 Sep 2003 01:28:14 -0400 From: John Kiernan Reply-To: jkiernan@uwo.ca To: "Behan, Rosemarie G" CC: "'histonet@lists.utsouthwestern.edu'" Subject: [Histonet] Glass cleaning for silver Rosemary Behan asked about cleaning glassware for silver staining. A reply follows. It's necessary to remove traces of metallic silver, which are not necessarily visible. The only common acid that does this is nitric. Here is what I do, with rationale. Let's assume the vessel is a Coplin jar. Rinse with one or two changes of distilled water. [Tap water contains chloride ions and silver chloride precipitation must be avoided.] Pour a few ml of concentrated nitric acid into the vessel and carefully let it wet all the inside surface. For a Coplin jar it's important to dissolve silver from all corners and from the slots for supporting slides. It takes 15-30 seconds to make black deposits or mirrors disappear. A minute of turning and tilting has to be enough to remove all the visible and invisible silver. Safely discard the nitric acid and fill up the vessel with distilled or otherwise purified water, three times. [Pure water must be used because tap water will precipitate traces of silver chloride from the residual nitric acid - which contains dissolved silver nitrate.] Wash in tap water, detergent etc as for any other dirty lab glassware, and don't spare the brush. [The nitric acid treatment does not remove all types of dirt. Bits of detached section, stuck to the glass, are made yellow by nitric acid.] Rinse in tap water, repeatedly, to get rid of the detergent (no more froth with shaking) and then in 3 generous changes of pure water to dilute out residual chloride from the tap water. Let the vessel dry by drainage and evaporation, then keep it in a closed cupboard, with its lid on (if it has a lid; and don't forget to clean the lid). If glassware is contaminated by insoluble silver compounds such as silver chloride, 5% sodium thiosulphate (10 minutes) will remove the silver. [The thiosulphate ion strongly complexes silver ions and will remove them from solid silver halides. This is the "fixation" of photographers.] In the above remarks I have not given detailed safety and disposal instructions. Conc. nitric acid is nasty stuff but becomes harmless when diluted with water. Do not use hydrochloric acid or an HCl-alcohol mixture for "acid washing" of glassware that will contain silver nitrate or protargol. [Reason is obvious from above discussion.] There are silver solvents less noxious than concentrated nitric acid. The best known one is Farmer's reducer. This is used in black & white photography for controlled removal of darkness (= silver) from negatives or prints. It is a solution containing potassium ferricyanide and sodium thiosulphate. Its actions on photo media take several minutes, but it takes much longer to weaken silver deposits on glassware and in overstained sections (my unpublished anecdotal observations). For what it's worth, I think concentrated nitric acid is the best cleaner of glassware used for silver methods. I also think that poor glass-hygiene (dishwashers etc etc etc) often causes failure or poor results with many staining techniques. -- ------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan@uwo.ca http://publish.uwo.ca/~jkiernan/ -- (Rosemary Behan: I've deleted many irrelevant messages from the tail of your email, which contained a megabyte of unrelated stuff. Please be careful about what to quote!) _____________________________________ "Behan, Rosemarie G" wrote: > > I am looking for a recipe for acid cleaning glassware to do silver stains, > can anyone help me? __________________________________________________ --__--__-- Message: 5 Date: Wed, 17 Sep 2003 09:50:57 +0200 From: "C.M. van der Loos" To: histonet@lists.utsouthwestern.edu Cc: v.lamanna@abdn.ac.uk Subject: [Histonet] RE: immunohistochemistry and frozen sections Dear Vincenzo, You didn't describe the fixative used for your frozen sections, but I assume that it was acetone. However, as you are trying to stain a nuclear marker it is strongly advised to use either Zamboni, 4% paraformaldehyde or just buffered formalin for 5 min at room temperature. Then wash with PBS (or TBS) and start your IHC procedure. Most nuclear markers get diffuse after acetone fixation. Furthermore, you described the application of different antigen retrieval methods. To my opinion, antigen retrieval is not needed for frozen sections because antigens/epitopes are not hidden due to fixation or something like with FFPE sections. Even if buffered formalin is used as fixative for frozen sections there is nothing to retrieve, simply because a 5 min fixation time is far too short to cause cross-linking of proteins. And indeed as you described, the tissue morphology of frozen sections will heavily suffer from antigen retrieval procedures. Good luck, Chris van der Loos Dept. of Cardiovascular Pathology Academical Medical Center Amsterdam - The Netherlands >Original Message ----- >From "Vincenzo La Manna" >Date Tue, 16 Sep 2003 15:48:43 +0100 (BST) >To >Subject [Histonet] immunohistochemistry and frozen sections > >Hi Histonetters, > >I've been struggling for 4 months on my 5 microns >frozen sections trying to find estrogen receptors. >I've tried both immunoperoxidase kit from Vector >laboratories either indirect immunofluorecsence (FITC >Goat anti mouse Secondary Ab from Sigma) but I've not >yet obtained good results. >My primary Ab is a mouse monoclonal to bovine >Estrogen receptor from Cymbus biotechnology. >Sections come from tissue that is supposed to be a >non-target tissue (mid laminar region from cow's >hooves)blocking with 2% goat normal serum (sigma) >whash in tbs buffer >All different methods for antigen retrieval have been >tested but sections look poor regarding histologic >quality and I still have very big problems with non >specific binding an staining (indirect >immunofluorescence). >It looks like frozen sections are too tender to >efford the whole processing. >Every suggestion or reference is welcome, >thank U > >La Manna Vincenzo >Department of Agriculture and Forestry, University of >Aberdeen, 581 King Street, AB24 5UA, Aberdeen , UK. >Telephone; 01224 274259 >Fax; 01224 273731 >e-mail v.lamanna@abdn.ac.uk --__--__-- Message: 6 From: "George Cole" To: Date: Wed, 17 Sep 2003 01:02:42 -0700 Subject: [Histonet] acid cleaning slides for silver preps This is a multi-part message in MIME format. ------=_NextPart_000_0001_01C37CB7.66CEBCB0 Content-Type: text/plain; charset="us-ascii" Content-Transfer-Encoding: 7bit Gray's Formulary contains a reliable cleaning procedure for slides: 40 parts saturated aqueous potassium dichromate Add, with due precautions, 60 parts sulfuric acid. I always just put excess dichromate in 60 parts water so that much solid dichromate remained in the bottom of a coverable Container. Add the 60 parts of Sulfuric Acid, with care. The quantity of cleaner may be a gallon or so, mixed in a container kept for that purpose. Put the coplin jars or other glass ware in the mixture for a few minutes or over night. Rinse the glass well with deionized water. This mixture can be used for a long time. But the best procedure is when the solid dichromate is gone, a fresh batch is mixed. ------=_NextPart_000_0001_01C37CB7.66CEBCB0 Content-Type: text/html; charset="us-ascii" Content-Transfer-Encoding: quoted-printable

Gray’s Formulary contains a reliable cleaning =3D procedure for slides:

40 parts saturated aqueous potassium = dichromate

Add, with due precautions, 60 parts sulfuric = acid.

I always just put excess dichromate in 60 parts water =3D so that much solid dichromate remained in the bottom of a coverable =

 Container.

Add the 60 parts of Sulfuric Acid, with = care.

The quantity of cleaner may be a gallon or so, mixed =3D in a container kept for that purpose.

Put the coplin jars or =3D other glass ware in the mixture for a few minutes or over night.  Rinse the glass well with deionized water.

This mixture can be used for a long time.  But the best procedure is when = the solid dichromate is gone, a fresh batch is mixed.  

------=_NextPart_000_0001_01C37CB7.66CEBCB0-- --__--__-- Message: 7 Date: Wed, 17 Sep 2003 09:22:38 +0100 From: "Mandy Townsend" To: "ANN MARUSKA" , Subject: RE: [Histonet] S100 frozen skin This is a multi-part message in MIME format. ------_=_NextPart_001_01C37CF4.DADB2C8C Content-Type: text/plain; charset="iso-8859-1" Content-Transfer-Encoding: quoted-printable Serotec are able to supply a polyclonal antibody against S100 that is =3D suitable for use on both cryostat and paraffin sections. Please do not =3D hesitate to contact me if you require further information. =20 Mandy =20 Mandy Townsend MSc=20 Technical Services Supervisor=20 Serotec Ltd=20 22 Bankside=20 Station Approach=20 Kidlington=20 Oxfordshire=20 OX5 1JE=20 Tel: +44 1865 852736=20 Fax: +44 1865 852739=20 email: mandy@serotec.co.uk=20 URL: www.serotec.com=20 Serotec-Your first choice for antibodies!=20 IMPORTANT NOTICE: This message and any attachments may be confidential. =3D If this has been sent to you in error, please contact the sender as soon =3D as possible. Serotec Ltd. Registered in England No.1604642=20 Registered Office: Boswell House, 1-5 Broad Street, Oxford, OX1 SAW. UK =3D -----Original Message----- From: ANN MARUSKA [mailto:amarusk1@FAIRVIEW.ORG] Sent: Tuesday, September 16, 2003 10:25 PM To: histonet@pathology.swmed.EDU Subject: [Histonet] S100 frozen skin Hi histonetters, =20 I have a researcher who is interested in doing an S100 on frozen skin =3D samples for melanoma....is anyone out there doing this? Do you know of =3D an S100 that works on frozen tissue? As always, thanks for your help. =20 Ann =20 Ann Maruska Fairview-University Medical Center Mpls. MN 55454 amarusk1@fairview.org 612-273-9119 ________________________________________________________________________ This e-mail has been scanned for all viruses by Star Internet. The service is powered by MessageLabs. For more information on a proactive anti-virus service working around the clock, around the globe, visit: http://www.star.net.uk ________________________________________________________________________ ------_=_NextPart_001_01C37CF4.DADB2C8C Content-Type: text/html; charset="iso-8859-1" Content-Transfer-Encoding: quoted-printable
Serotec are able to supply a polyclonal antibody against = S100 that=20 is suitable for use on both cryostat and paraffin sections.  Please =3D do not=20 hesitate to contact me if you require further = information.
 
Mandy
 

Mandy Townsend MSc =3D
Technical Services Supervisor
=3D
Serotec Ltd =
22 Bankside
Station Approach
Kidlington
Oxfordshire
OX5 1JE=20

Tel: +44 1865 852736 =3D
Fax: +44 1865 852739
email: mandy@serotec.co.uk =3D
URL: www.serotec.com
Serotec-Your = first choice for=20 antibodies!

IMPORTANT NOTICE: This message = and any=20 attachments may be confidential.  If this has been sent to you in =3D error,=20 please contact the sender as soon as possible.

Serotec Ltd. Registered in =3D England=20 No.1604642
Registered = Office:=20 Boswell House, 1-5  Broad Street, Oxford, OX1 SAW. UK=20

-----Original Message-----
From: ANN MARUSKA=20 [mailto:amarusk1@FAIRVIEW.ORG]
Sent: Tuesday, September 16, =3D 2003 10:25=20 PM
To: histonet@pathology.swmed.EDU
Subject: = [Histonet] S100=20 frozen skin

Hi histonetters,
 
I have a researcher who is interested in doing an S100 on frozen =3D skin=20 samples for melanoma....is anyone out there doing this?  Do you = know of an=20 S100 that works on frozen tissue?
As always, thanks for your help.
 
Ann
 
Ann Maruska
Fairview-University Medical Center
Mpls. MN  =3D 55454
amarusk1@fairview.org
612-27= 3-9119

_________________________________________________________=3D _______________
This=20 e-mail has been scanned for all viruses by Star Internet. The
service =3D is=20 powered by MessageLabs. For more information on a = proactive
anti-virus=20 service working around the clock, around the globe, visit:
http://www.star.net.uk
___________= _____________________________________________________________
<=3D /HTML> ------_=_NextPart_001_01C37CF4.DADB2C8C-- --__--__-- Message: 8 From: "peptolab" To: Date: Wed, 17 Sep 2003 04:58:06 -0400 Subject: [Histonet] Yarrow Common yarrow, or Achillea millefolium, spreads rapidly from underground rhizomes - plants placed two feet apart will fill in within one year. The mat-like, dark green finely divided ferny foliaged plants will take over any ground available and likes sunny average soil not partcularly rich. It is relatively drought tolerant. It is not a good border plant but is very useful for sunny dry waste places where a flowering groundcover would be useful. This achillea and A. ptarmica are invasive while the other species such as A. filipendulina, grandifolia, and tomentosa are well behaved clump formers, often with attractive glaucous (blue gray) and fuzzy foliage. Achillea has been used as a toothache remedy in Europe (1440) and was mixed in ale instead of hops to increase inebriation. It was said to grow in churchyards as a a reproach to the dead who "might not have died had they taken their daily yarrow", and was used to heal wounds- Achilles (Achillea) supposedly used it to stauch the wound of his soldiers. I am unaware of any histological use for the herb, except perhaps to staunch the wounds of disposable blades or broken lurking coverslip fragments. Jeff Silverman- Plantsman Southside Hospital Bay Shore NY --__--__-- Message: 9 From: "Mezey Szilvia" To: histonet@lists.utsouthwestern.edu Date: Wed, 17 Sep 2003 13:53:58 +0200 Subject: [Histonet] BrdU and EM? Hello All, I'm trying to do BrdU staining (in paraformaldehyde fixed, free- floating sections) and preserve the ultrastructure of the tissue for EM at the same time. HCl works fine for denaturing DNA for BrdU- ICC but doesn't leave much of the tissue for EM. DNase would be more EM-friendly but doesn't penetrate the tissue enough. Does anybody know a method that could provide a fair enough compromise between BrdU and EM? Maybe by increasing the penetration of DNase? Best regards to you all, Szilvi Szilvia Mezey PhD student Semmelweis University Dept. of Anatomy, Histology and Embryology Tuzolto u. 58. Budapest, 1094, Hungary T.: +36-12156920/3687 F.: +36-12155158 E-mail: mezey@ana.sote.hu --__--__-- Message: 10 From: "Priscilla Delventhal" To: histonet@lists.utsouthwestern.edu Date: Wed, 17 Sep 2003 07:00:18 -0600 Subject: [Histonet] job openings in Dallas Hi Tom - I will be in the San Antonio area starting next week for about a month. I know Dallas is across the state but thought I'd tell you what my plans are. I am retiring from full time work this week and plan on taking a vacation for the rest of the month. After that I only want to do temp openings - a couple a year. I have 35+ years experience in histology, both supervisory and bench. I have my BA, and HT and HTL. Let me know if you could use some temporary help this Fall. My cell phone is 307-851-5595. I will be checking messages a couple of times a day for the rest of this week and then will have it on full time as I travel down to San Antonio. Hoping to hear from you. You make your lab sound wonderful. Priscilla Delventhal --__--__-- Message: 11 To: histonet@lists.utsouthwestern.edu From: Joyce Christopher Date: Wed, 17 Sep 2003 08:10:56 -0500 Subject: [Histonet] KnifeMaker Our Reichert-Jung Knifemaker II has developed a problem. It no longer has the strength to break the knives. In other words its spring needs replacement. Problem - the spring part is no longer available. Does anyone have a knifemaker they are no longer using or has one that is no longer working but does have a working spring? Joyce Christopher Bayer CropScience LP 17745 S. Metcalf Stilwell, KS 66085 913-433-5244 joyce.christopher@bayercropscience.com --__--__-- Message: 12 From: Amy Self To: "'histonet@lists.utsouthwestern.edu'" Date: Wed, 17 Sep 2003 09:36:14 -0400 Subject: [Histonet] receiving un-registered specimens Hi all Netters, I was wandering how everyone rec'd specimens in the histology lab? Are they registered into the hospital system before they are grossed in or are they brought straight to the histo lab, grossed and then registered. We have an on-going problem with specimens that are coming to the histology lab before they are getting registered in the hospital system. This causes a lot of confusion with everyone. Also does anyone have a specific policy for handling formalin spills that they could share with me? We have a general policy for lab spills but I don't think that it is detailed enough for the histology lab. Thanks for all the help that you all send out not only to me but to everyone.....Much appreciated. Amy Self Georgetown Hospital Systems 843-527-7179 (home) 843-520-7882 (fax) Note: The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. --__--__-- Message: 13 From: "Mezey Szilvia" To: "Charles W. Scouten, Ph.D." , histonet@lists.utsouthwestern.edu Date: Wed, 17 Sep 2003 15:44:09 +0200 Subject: RE: [Histonet] BrdU and EM? Animal tissue (domestic chicks), transcardially perfused. Szilvi Subject: RE: [Histonet] BrdU and EM? Date sent: Wed, 17 Sep 2003 08:03:03 -0500 From: "Charles W. Scouten, Ph.D." To: "Mezey Szilvia" > Animal tissue (perfused?) or human tissue? > > Charles W.=A0 Scouten, Ph.D. > myNeuroLab.com > 5918 Evergreen Blvd. > St. Louis, MO 63134 > Ph: 314 522 0300=A0 > FAX=A0 314 522 0377 > cwscouten@myneurolab.com > www.myneurolab.com > > > -----Original Message----- > From: Mezey Szilvia [mailto:mezey@ana.sote.hu] > Sent: Wednesday, September 17, 2003 6:54 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] BrdU and EM? > > Hello All, > > I'm trying to do BrdU staining (in paraformaldehyde fixed, free- > floating sections) and preserve the ultrastructure of the tissue for > EM at the same time. HCl works fine for denaturing DNA for BrdU- > ICC but doesn't leave much of the tissue for EM. DNase would be > more EM-friendly but doesn't penetrate the tissue enough. > > Does anybody know a method that could provide a fair enough > compromise between BrdU and EM? Maybe by increasing the > penetration of DNase? > > Best regards to you all, > > Szilvi > > > Szilvia Mezey > PhD student > Semmelweis University > Dept. of Anatomy, Histology and Embryology > Tuzolto u. 58. Budapest, 1094, Hungary > T.: +36-12156920/3687 > F.: +36-12155158 > E-mail: mezey@ana.sote.hu > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet Szilvia Mezey PhD student Semmelweis University Dept. of Anatomy, Histology and Embryology Tuzolto u. 58. Budapest, 1094, Hungary T.: +36-12156920/3687 F.: +36-12155158 E-mail: mezey@ana.sote.hu --__--__-- Message: 14 From: "Dawson, Glen" To: histonet@lists.utsouthwestern.edu Date: Wed, 17 Sep 2003 08:42:50 -0500 Subject: [Histonet] wage/vacancy survey All, Does anyone know the release date for the official 2002 ASCP wage/vacancy survey? I have seen the preliminary survey and I seem to remember someone mentioning that the complete survey would be released in September. Any info would be appreciated. Thanx, Glen Dawson --__--__-- Message: 15 From: DJStashev@aol.com Date: Wed, 17 Sep 2003 09:45:16 EDT To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Used JB-4 Microtome? I am trying to locate a used JB-4 Sorvall microtome to buy. Does anyone out in Histo-land have this type of microtome that they are no longer using??? Thanks, Jen Stashevsky I.U. Medical Center 317-576-0338 --__--__-- Message: 16 Date: Wed, 17 Sep 2003 08:09:27 -0600 To: Joyce Christopher , Histonet@lists.utsouthwestern.edu From: Gayle Callis Subject: Re: [Histonet] KnifeMaker See if you can get a local metalworking shop (we have a fellow who does scientific instruments) make one for you or out of some type of springlike material and get several backups. Hopefully, Reichert Jung (did you try contacting Leica Microsystems about the problem?) might have one in a drawer somewhere. Contact Don.Birgerson@leica-microsystems.com, he is very knowledgable about these instruments, and he has always been graciously helpful with plights like this. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) --__--__-- Message: 17 Date: Wed, 17 Sep 2003 10:28:59 -0400 From: "Fred Underwood" To: Subject: Re: [Histonet] questions regarding to a-Naphthyl Butyrate Esterase stain and a-Naphthyl Acetate stain(no I have used the kits put together by Sigma. The results were good. However, strict adherence to expiration dates, temperatures, times and solution prepatation is necessary. Otherwise the results can be variable and less than optimal. Fred >>> weiping Ren 09/16/03 01:13PM >>> Hi, Histonetters: I need to set up non specific esterase staining method on bone marrow smear samples. My questions are: 1. Are a-Naphthyl Butyrate Esterase stain and a-Naphthyl Acetate stain(non specific esterase) are the same stain? 2. If these are two different stains but have the same clinical diagnosis significance, which one is better regarding to the time and accuracy? 3. Are there any protocols for these stains? Where I can get these references? Thank you! Any suggestions and advice welcome. Weiping Wayne State University Detroit, MI --------------------------------- Do you Yahoo!? Yahoo! SiteBuilder - Free, easy-to-use web site design software --__--__-- Message: 18 Date: Wed, 17 Sep 2003 10:32:22 -0400 From: "Claye Clyatt" To: , Subject: Re: [Histonet] microwave Personally, I find that microwaving in general creates more problems than = they solve. We no longer use one for our laboratory procedures. Claye Claye Clyatt Chief Histotechnologist Department of Pathology=20 Room #BF119 Medical College of Georgia Augusta, Ga 30912 office (706) 721-3630 pager (706) 721-7243-1132 e-mail: cclyatt@mail.mcg.edu >>> "Gudrun Lang" 09/16/03 05:14PM >>> Hi Histonetters! I have a question about microwave oven in histolabs. We did introduce such = a household thing for IHC, but it was not very successfull. Now we heat =3D our tea with it. IHC works with heat and steam. Which equipment do you use for special staining? Are there specific = apparats or also the houshold ones? And has it become really common to =3D work with it? thanks for your answers. Gundi Lang general hospital Linz, Austria --__--__-- Message: 19 Date: Wed, 17 Sep 2003 10:34:37 -0400 From: "Claye Clyatt" To: , Subject: Re: [Histonet] Acetylcholonesterase Try Sigma. I've used their kits in the past and they work very nice and =3D are easy to use. Claye >>> 09/16/03 07:02PM >>> Anyone have an easy recipe (or refer me to a prepared kit that can be=20 purchased) for the acetylcholonesterase stain? Doctor is looking for =3D Hirschsprungs=20 disease. Any help is appreciated. Sincerely, Deb King, HT (ASCP) Sacramento, CA --__--__-- Message: 20 From: "Pamela Marcum" To: "Fred Underwood" , Date: Wed, 17 Sep 2003 10:38:48 -0400 Subject: RE: [Histonet] questions regarding to a-Naphthyl ButyrateEsterase stain and a-Naphthyl Acetate stain(no and filter paper requirements should be followed use exactly what they say or risk the reaction hydrolyzing in the filter. Pam Marcum > -----Original Message----- > From: histonet-admin@lists.utsouthwestern.edu > [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of Fred > Underwood > Sent: Wednesday, September 17, 2003 10:29 AM > To: histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] questions regarding to a-Naphthyl > ButyrateEsterase stain and a-Naphthyl Acetate stain(no > > > I have used the kits put together by Sigma. The results were good. > However, strict adherence to expiration dates, temperatures, times and > solution prepatation is necessary. Otherwise the results can be > variable and less than optimal. > > Fred > > >>> weiping Ren 09/16/03 01:13PM >>> > Hi, Histonetters: > I need to set up non specific esterase staining method on bone marrow > smear samples. > > My questions are: > 1. Are a-Naphthyl Butyrate Esterase stain and a-Naphthyl Acetate > stain(non specific esterase) are the same stain? > 2. If these are two different stains but have the same clinical > diagnosis significance, which one is better regarding to the time and > accuracy? > 3. Are there any protocols for these stains? Where I can get these > references? > > Thank you! > > Any suggestions and advice welcome. > > Weiping > Wayne State University > Detroit, MI > > > > --------------------------------- > Do you Yahoo!? > Yahoo! SiteBuilder - Free, easy-to-use web site design software > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > --__--__-- Message: 21 From: "Pamela Marcum" To: "Fred Underwood" , Date: Wed, 17 Sep 2003 10:38:48 -0400 Subject: RE: [Histonet] questions regarding to a-Naphthyl ButyrateEsterase stain and a-Naphthyl Acetate stain(no and filter paper requirements should be followed use exactly what they say or risk the reaction hydrolyzing in the filter. Pam Marcum > -----Original Message----- > From: histonet-admin@lists.utsouthwestern.edu > [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of Fred > Underwood > Sent: Wednesday, September 17, 2003 10:29 AM > To: histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] questions regarding to a-Naphthyl > ButyrateEsterase stain and a-Naphthyl Acetate stain(no > > > I have used the kits put together by Sigma. The results were good. > However, strict adherence to expiration dates, temperatures, times and > solution prepatation is necessary. Otherwise the results can be > variable and less than optimal. > > Fred > > >>> weiping Ren 09/16/03 01:13PM >>> > Hi, Histonetters: > I need to set up non specific esterase staining method on bone marrow > smear samples. > > My questions are: > 1. Are a-Naphthyl Butyrate Esterase stain and a-Naphthyl Acetate > stain(non specific esterase) are the same stain? > 2. If these are two different stains but have the same clinical > diagnosis significance, which one is better regarding to the time and > accuracy? > 3. Are there any protocols for these stains? Where I can get these > references? > > Thank you! > > Any suggestions and advice welcome. > > Weiping > Wayne State University > Detroit, MI > > > > --------------------------------- > Do you Yahoo!? > Yahoo! SiteBuilder - Free, easy-to-use web site design software > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > --__--__-- Message: 22 From: "Pamela Marcum" To: "Fred Underwood" , Date: Wed, 17 Sep 2003 10:38:48 -0400 Subject: RE: [Histonet] questions regarding to a-Naphthyl ButyrateEsterase stain and a-Naphthyl Acetate stain(no and filter paper requirements should be followed use exactly what they say or risk the reaction hydrolyzing in the filter. Pam Marcum > -----Original Message----- > From: histonet-admin@lists.utsouthwestern.edu > [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of Fred > Underwood > Sent: Wednesday, September 17, 2003 10:29 AM > To: histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] questions regarding to a-Naphthyl > ButyrateEsterase stain and a-Naphthyl Acetate stain(no > > > I have used the kits put together by Sigma. The results were good. > However, strict adherence to expiration dates, temperatures, times and > solution prepatation is necessary. Otherwise the results can be > variable and less than optimal. > > Fred > > >>> weiping Ren 09/16/03 01:13PM >>> > Hi, Histonetters: > I need to set up non specific esterase staining method on bone marrow > smear samples. > > My questions are: > 1. Are a-Naphthyl Butyrate Esterase stain and a-Naphthyl Acetate > stain(non specific esterase) are the same stain? > 2. If these are two different stains but have the same clinical > diagnosis significance, which one is better regarding to the time and > accuracy? > 3. Are there any protocols for these stains? Where I can get these > references? > > Thank you! > > Any suggestions and advice welcome. > > Weiping > Wayne State University > Detroit, MI > > > > --------------------------------- > Do you Yahoo!? > Yahoo! SiteBuilder - Free, easy-to-use web site design software > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > --__--__-- Message: 23 Date: Wed, 17 Sep 2003 11:26:33 -0400 From: "Vicki Gauch" To: , Subject: Re: [Histonet] receiving un-registered specimens Amy, We receive specimens that are already registered into the Hospital System and also ones that need to be registered. The first set are easily handled as they are just accessioned into the system and processed as normal. The second set are sent down the hall to registration to receive a medical record and account number and then we pick the reqs up and bring them back to our lab to be accessioned and processed. As for the formalin spill policy, we use Green Z on small spills and have a procedure for that....large spills we would call our Environmental Health and Safety Unit to come and clean up the spill and we have a procedure in place for that. Hope that helps, Vicki Gauch AMCH Pathology Albany, NY >>> Amy Self 9/17/2003 9:36:14 AM >>> Hi all Netters, I was wandering how everyone rec'd specimens in the histology lab? Are they registered into the hospital system before they are grossed in or are they brought straight to the histo lab, grossed and then registered. We have an on-going problem with specimens that are coming to the histology lab before they are getting registered in the hospital system. This causes a lot of confusion with everyone. Also does anyone have a specific policy for handling formalin spills that they could share with me? We have a general policy for lab spills but I don't think that it is detailed enough for the histology lab. Thanks for all the help that you all send out not only to me but to everyone.....Much appreciated. Amy Self Georgetown Hospital Systems 843-527-7179 (home) 843-520-7882 (fax) Note: The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --__--__-- Message: 24 From: "Morken, Tim - Labvision" To: 'Amy Self' , "'histonet@lists.utsouthwestern.edu'" Date: Wed, 17 Sep 2003 11:30:16 -0400 Subject: RE: [Histonet] receiving un-registered specimens I have never received a gross specimen in histology that was not from a registered patient. We did receive blocks and slides for referral cases that we had to put into the system. Tim Morken -----Original Message----- From: Amy Self [mailto:ASelf@gmhsc.com] Sent: Wednesday, September 17, 2003 6:36 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] receiving un-registered specimens Hi all Netters, I was wandering how everyone rec'd specimens in the histology lab? Are they registered into the hospital system before they are grossed in or are they brought straight to the histo lab, grossed and then registered. We have an on-going problem with specimens that are coming to the histology lab before they are getting registered in the hospital system. This causes a lot of confusion with everyone. Also does anyone have a specific policy for handling formalin spills that they could share with me? We have a general policy for lab spills but I don't think that it is detailed enough for the histology lab. Thanks for all the help that you all send out not only to me but to everyone.....Much appreciated. Amy Self Georgetown Hospital Systems 843-527-7179 (home) 843-520-7882 (fax) Note: The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --__--__-- Message: 25 From: "Pamela Marcum" To: "Fred Underwood" , Date: Wed, 17 Sep 2003 10:38:48 -0400 Subject: RE: [Histonet] questions regarding to a-Naphthyl ButyrateEsterase stain and a-Naphthyl Acetate stain(no and filter paper requirements should be followed use exactly what they say or risk the reaction hydrolyzing in the filter. Pam Marcum > -----Original Message----- > From: histonet-admin@lists.utsouthwestern.edu > [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of Fred > Underwood > Sent: Wednesday, September 17, 2003 10:29 AM > To: histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] questions regarding to a-Naphthyl > ButyrateEsterase stain and a-Naphthyl Acetate stain(no > > > I have used the kits put together by Sigma. The results were good. > However, strict adherence to expiration dates, temperatures, times and > solution prepatation is necessary. Otherwise the results can be > variable and less than optimal. > > Fred > > >>> weiping Ren 09/16/03 01:13PM >>> > Hi, Histonetters: > I need to set up non specific esterase staining method on bone marrow > smear samples. > > My questions are: > 1. Are a-Naphthyl Butyrate Esterase stain and a-Naphthyl Acetate > stain(non specific esterase) are the same stain? > 2. If these are two different stains but have the same clinical > diagnosis significance, which one is better regarding to the time and > accuracy? > 3. Are there any protocols for these stains? Where I can get these > references? > > Thank you! > > Any suggestions and advice welcome. > > Weiping > Wayne State University > Detroit, MI > > > > --------------------------------- > Do you Yahoo!? > Yahoo! SiteBuilder - Free, easy-to-use web site design software > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > --__--__-- Message: 26 Date: Wed, 17 Sep 2003 12:24:46 -0400 From: "Richard Cartun" To: Subject: [Histonet] Ret IHC We are do "Ret" immunoperoxidase staining on thryoid tissue using a rabbit polyclonal antibody from Santa Cruz (sc-167). Does anyone have experience with this antibody on formalin-fixed, paraffin-embedded tissue? Thank you. Richard Cartun --__--__-- Message: 27 From: "Michelle D. Moore" To: "Histonet" Date: Wed, 17 Sep 2003 10:30:47 -0600 Organization: The Memorial Hospital Subject: [Histonet] Underprocessed tissue This is a multi-part message in MIME format. ------=_NextPart_000_0042_01C37D06.C21779A0 Content-Type: text/plain; charset="iso-8859-1" Content-Transfer-Encoding: quoted-printable Hello fellow histonetters, hope everyone is having a good day so far! I =3D am in a bind and I know you can help me, I was embedding this morning =3D and having a very difficult time with my tissues, so I finally finished =3D embedding and when I started to cut, nothing and I mean nothing would =3D section UUGH. Well in the process I thought maybe it was some new = paraffin I was trying so I didn't get to excited until I couldn't cut =3D anything. Needless to say what I need from you wonderful people is help =3D on reprocessing my tissue I know I need to remove the paraffin that's in =3D the tissue and clear the xylene out but I have no procedure for doing =3D this and it has been years since I have had to do it. Any help you could =3D direct my way or any procedure that you would be willing to share would =3D be greatly appreciated!! THANK YOU in advance for saving my rear I do =3D appreciate it a lot. That's the beauty of the histonet people helping =3D people. Michelle D. Moore HT (ASCP) The Memorial Hospital Craig, CO ------=_NextPart_000_0042_01C37D06.C21779A0 Content-Type: text/html; charset="iso-8859-1" Content-Transfer-Encoding: quoted-printable
Hello fellow histonetters, hope = everyone is having=20 a good day so far! I am in a bind and I know you can help me, I was = embedding=20 this morning and having a very difficult time with my tissues, so I = finally=20 finished embedding and when I started to cut, nothing and I mean nothing =3D would=20 section UUGH. Well in the process I thought maybe it was some new = paraffin I was=20 trying so I didn't get to excited until I couldn't cut anything. = Needless to say=20 what I need from you wonderful people is help on reprocessing my tissue =3D I know I=20 need to remove the paraffin that's in the tissue and clear the xylene =3D out but I=20 have no procedure for doing this and it has been years since I have had =3D to do=20 it. Any help you could direct my way or any procedure that you would be =3D willing=20 to share would be greatly appreciated!! THANK YOU in advance for saving =3D my rear=20 I do appreciate it a lot. That's the beauty of the histonet people = helping=20 people.
 
Michelle D. Moore HT = (ASCP)
The Memorial Hospital
Craig, CO
------=_NextPart_000_0042_01C37D06.C21779A0-- --__--__-- Message: 28 Date: Wed, 17 Sep 2003 12:42:20 -0400 From: Andrew Fedanov To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Clot sections Hi Histonetters, Does anybody has an experience in preparation of clot sections? Thanks -- Andrew Fedanov Ph.D. Emory Vaccine Center at Yerkes 954 Gatewood Rd. NE Atlanta, GA 30329 Phone: 404-727-3043 Fax: 404-727-8199 --__--__-- Message: 29 Date: Wed, 17 Sep 2003 12:59:04 -0400 From: DDittus787@aol.com To: Rcartun@harthosp.org, histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Ret IHC Rich: we currently use c-ret , we do antigen retrieval in citrate, control slides must be fresh and incubate 32 minutes at 42 degrees Dana --__--__-- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet

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