RE: [Histonet] Osteoclast
You can do a TRAP stain on EDTA decaled bone. I haven't been able to get it to work on formic acid decaled bone. I also believe that there is an antibody for osteoclasts, but I have not tried it. Loralee McMahon, HTL (ASCP) Immunohistochemistry Supervisor Strong Memorial Hospital Department of Surgical Pathology (585) 275-7210 From: histonet-boun...@lists.utsouthwestern.edu [histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Lin Bustamante [lbustama...@cvm.tamu.edu] Sent: Tuesday, October 05, 2010 4:58 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Osteoclast We need to find a way to stain mainly Osteoclast. Any suggestions? Thank you very much. Lin Lin S. Bustamante, B.Sc.; HT(ASCP) Research Associate Histology Lab Supervisor Veterinary Integrative Bioscience Texas AM University College Station, TX 77843-4458 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Question about Oil Red O controls
Hello Histonetters, I am trying to find a procedure for using butter and egg yolks as controls for the Oil Red O stain (to show the fat). Does anyone have something they would be able to share with me? Thanks, Komal ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Osteoclast
Hi Loralee, I am having similar problems with Formic acid Decal'd sections for TRAP. After EDTA, are you doing a TRAP IHC or the enzyme assay from Sigma? As for molecular markers, Cathepsin K is pretty good for Osteoclasts but I do westerns in a research setting..has anyone had good success with any antibody for immunostaining? Best, Praveen Arany, Graduate Student, Harvard University, Cambridge MA On 10/6/2010 8:43 AM, McMahon, Loralee A wrote: You can do a TRAP stain on EDTA decaled bone. I haven't been able to get it to work on formic acid decaled bone. I also believe that there is an antibody for osteoclasts, but I have not tried it. Loralee McMahon, HTL (ASCP) Immunohistochemistry Supervisor Strong Memorial Hospital Department of Surgical Pathology (585) 275-7210 From: histonet-boun...@lists.utsouthwestern.edu [histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Lin Bustamante [lbustama...@cvm.tamu.edu] Sent: Tuesday, October 05, 2010 4:58 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Osteoclast We need to find a way to stain mainly Osteoclast. Any suggestions? Thank you very much. Lin Lin S. Bustamante, B.Sc.; HT(ASCP) Research Associate Histology Lab Supervisor Veterinary Integrative Bioscience Texas AM University College Station, TX 77843-4458 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Question about Oil Red O controls
Just use Mayo (not fat free, of course!) and smear it on the slide like you would a blood smear. It stains beautifully. Drew On Wed, Oct 6, 2010 at 09:15, Komal Gada kjg...@gmail.com wrote: Hello Histonetters, I am trying to find a procedure for using butter and egg yolks as controls for the Oil Red O stain (to show the fat). Does anyone have something they would be able to share with me? Thanks, Komal ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Osteoclast
We were never able to get the TRAP working with Formic Acid decal. Using a home brewed TRAP kit or the Sigma Kit. But Cell Marque has a TRAcP antibody (catalog 341M-95) that is used as a hairy cell marker. But it also stains macrophages and osteoclasts. Loralee McMahon, HTL (ASCP) Immunohistochemistry Supervisor Strong Memorial Hospital Department of Surgical Pathology (585) 275-7210 From: histonet-boun...@lists.utsouthwestern.edu [histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Praveen Arany [ar...@fas.harvard.edu] Sent: Wednesday, October 06, 2010 9:17 AM To: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Osteoclast Hi Loralee, I am having similar problems with Formic acid Decal'd sections for TRAP. After EDTA, are you doing a TRAP IHC or the enzyme assay from Sigma? As for molecular markers, Cathepsin K is pretty good for Osteoclasts but I do westerns in a research setting..has anyone had good success with any antibody for immunostaining? Best, Praveen Arany, Graduate Student, Harvard University, Cambridge MA On 10/6/2010 8:43 AM, McMahon, Loralee A wrote: You can do a TRAP stain on EDTA decaled bone. I haven't been able to get it to work on formic acid decaled bone. I also believe that there is an antibody for osteoclasts, but I have not tried it. Loralee McMahon, HTL (ASCP) Immunohistochemistry Supervisor Strong Memorial Hospital Department of Surgical Pathology (585) 275-7210 From: histonet-boun...@lists.utsouthwestern.edu [histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Lin Bustamante [lbustama...@cvm.tamu.edu] Sent: Tuesday, October 05, 2010 4:58 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Osteoclast We need to find a way to stain mainly Osteoclast. Any suggestions? Thank you very much. Lin Lin S. Bustamante, B.Sc.; HT(ASCP) Research Associate Histology Lab Supervisor Veterinary Integrative Bioscience Texas AM University College Station, TX 77843-4458 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] RE: embedding beads/tissue marking dyes
TBS has good dyes...you can mordant them (what Bouins does) in fo
rmalin-aceto-alcohol. This is less toxic then Bouins and works
well.= nbsp; Of course if you are doing anything with breast tissue,
you don't wan= t to leave it in there but a second or two because it
is not known if it ef= fects ER/PR results.
Happy Fixing!!
PS-W= e called F-A-A juicy juice because that's what it smells like
=)
Sarah = Goebel, B.A., HT (ASCP)
Histotechnician
= div
XBiot= ech USA Inc.
8201 East Riverside Dr. Bldg 4 Suite 100
Austin, Texas 78744
[DEL: (512)386-5107
:DEL]
Original Message
Subject: [Histonet] RE: embedding beads/tissue marking dyes
From: Maria Katleba [1]Maria.Ka= tl...@stjoe.org
Date: Tue, October 05, 2010 2:40 pm
To: Jacqueline Farnsworth [2]jacqueline.farnswo...@cls.ab.ca,
[3]histo...@lists.utsout= hwestern.edu [4]
Histonet@lists.utsouthwestern.edu
When you place dye on any tissue, make sure you pipette some bouins on
to t= he tissue... Apparently the bouins sets the stain...
afterwards dab the t= issue with paper towel to sop up the excess
dye/bouins.
Some dyes are better than others, but try the bouins first
Call me or email me directly, I can help you with details :)
Maria Katleba MS HT(ASCP)
Pathology Dept. Mgr
Queen of the Valley Medical Center
1000 Trancas Street
Napa CA 94558
(707) 252-4411 x3689 direct
(707) 226-4385 pager
(707) 294-9229 cell- anytime
-Original Message-
From: [5]histonet= -boun...@lists.utsouthwestern.edu
[[6]mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of
Jacqueline Farnsworth
Sent: Tuesday, October 05, 2010 2:14 PM
To: [7]histo...@lists.uts= outhwestern.edu
Subject: [Histonet] embedding beads/tissue marking dyes
Hi
We may be encountering some issues with our tissue marking dyes
'clogging' = up our processors. Ive heard of a system where a
coloured 'bead' is embed= ded right beside the tissue, and
subsequently cut onto the slide. This doe= s not mark the margins
obviously, but is used as a method to track like spe= cimens that are
grossed, embedded and subsequently cut in a row. (red, oran= ge,
green, blue.)
I explored the archives for embedding beads, but only found
reference to = a glass bead that is placed in the wax inside the
block, but not subsequent= ly cut.
PS: we are still troubleshooting our dyes (dilutions, brand, etc.),
but if = anyone has a brand of dye that they like and have no issues
with, I'd be th= rilled to get the information as well!
Thanks in advance,
Jacquie
Jacqueline Farnsworth
Anatomic Pathology, Tech III
Diagnostic Scientific Centre
Calgary Laboratory Services
Phone: 403-770-3588
Pager: 403-212-8223 X07630
P Please consider the environment before printing this email.
This message and any attached documents are only for the use of the
intende= d recipient(s), are confidential and may contain privileged
information. An= y unauthorized review, use, retransmission, or other
disclosure is strictly= prohibited. If you have received this message
in error, please notify the = sender immediately, and then delete the
original message. Thank you.
___
Histonet mailing list
[8]histo...@lists.utsouth= western.edu
[9]http:= //lists.utsouthwestern.edu/mailman/listinfo/histonet
Notice from St. Joseph Health System:
Please note that the information contained in this message may be
privilege= d and confidential and protected from disclosure.
___
Histonet mailing list
[10]histo...@lists.utsouth= western.edu
[11]http:= //lists.utsouthwestern.edu/mailman/listinfo/histonet
References
1. 3Dmailto:maria.katl...@stjoe.org;
2. 3Dmailto:jacqueline.farnswo...@cls.a 3.
3Dmailto:Histonet@lists.utsouthwestern.edu;
4. 3Dmailto:Histonet@lists.utsouthwestern.edu;
5. 3Dmailto:histonet-boun...@lists.utsouthwestern.edu;
6. 3Dmailto:histonet-boun...@l 7.
3Dmailto:Histonet@lists.utsouthwestern.edu;
8. 3Dmailto:Histonet@lists.utsouthwestern.edu;
9. 3Dhttp://lists.utsouthwestern.edu/mailman/listinfo/histonet;
10. 3Dmailto:Histonet@lists.utsouthwestern.edu;
11. 3Dhttp://lists.utsouthwestern.edu/mailman/listinfo/histonet;
___
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Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] AChE fiber staining protocoll?
I have had great success in fine fibers with the modification of Martucciello et al, Eur J Pediatr Surg 2001; 11: 300-304, but only in 15 micron intestine sections, so I do not know how it will transpose to brain studies and the thicker sections Ronnie Houston Anatomic Pathology Manager Nationwide Children's Hospital Columbus OH 43205 (614) 722 5450 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of szi...@bio.u-szeged.hu Sent: Wednesday, October 06, 2010 3:34 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] AChE fiber staining protocoll? Dear Histonet Members! We are going to investigate AChE fiber density changes in the cortex of transcardially perfused rat brain slices (30 micrometer, criostat sections, perfusing solution: 4% formaldehyde in PB ph 7.4). We need a protocoll to visualize FINE fiber structure to be able to count changes. We used modification of Hedreen and original Tago protocoll. Hedreen good results for general, but no fiber staining, Tago no sucsses. Could you please tell us some advise, protocolls functioning on teh above mentioned objects? Thank you all in advance... Csaba Szigeti This message was sent using IMP, the Internet Messaging Program. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet - Confidentiality Notice: The following mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. The recipient is responsible to maintain the confidentiality of this information and to use the information only for authorized purposes. If you are not the intended recipient (or authorized to receive information for the intended recipient), you are hereby notified that any review, use, disclosure, distribution, copying, printing, or action taken in reliance on the contents of this e-mail is strictly prohibited. If you have received this communication in error, please notify us immediately by reply e-mail and destroy all copies of the original message. Thank you. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] FW: goat anti-rabbit FITC
Hi, Could anyone please help us out? Does anyone use goat anti-rabbit FITC from Abcam? We would like to know the dilution used any other hints you have. We will be using it on the Dako autostainer to detect rabbit C4d in renal frozen sections. I am sending this on behalf or our IHC lab, please respond to: dwat...@hsc.mb.ca or to me I can pass it along. Thanks for any help you can provide. Sharon Allen HSC, Wpg, MB, Ca This email and/or any documents in this transmission is intended for the addressee(s) only and may contain legally privileged or confidential information. Any unauthorized use, disclosure, distribution, copying or dissemination is strictly prohibited. If you receive this transmission in error, please notify the sender immediately and return the original. Ce courriel et tout document dans cette transmission est destiné à la personne ou aux personnes à qui il est adressé. Il peut contenir des informations privilégiées ou confidentielles. Toute utilisation, divulgation, distribution, copie, ou diffusion non autorisée est strictement défendue. Si vous n'êtes pas le destinataire de ce message, veuillez en informer l'expéditeur immédiatement et lui remettre l'original. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Helicobacter pylori
Good Morning Histo land. I would like to know what others are using for a special stain for Helicobacter pylori not IHC. Do you use a kit? From where? Make up your own? Procedure. Thanks you. You have always been so helpful. Kathy Gorham H.T. GRH National Recognition Outstanding Rural Health Organization of 2009 awarded by NRHA Gold Standard Critical Access Hospital 2009 awarded by LarsonAllen LLP Leader in Innovative Excellence 2009 awarded by the OAHHS Financial Excellence Award 2010 awarded by the national Rural Health Research Policy Analysis Center Healthcare Achievement Award for Quality in Patient Care Delivery and Satisfaction 2010 awarded by Amerinet GRH Mission We will ensure access to high-quality, cost-effective health services in a safe, customer-friendly environment for all those in need of our services. GRH Confidentiality Notice This e-mail and any attached documents are for the intended recipient/s only and should be protected against viewing by unauthorized persons. The information herein may have been disclosed from records whose confidentiality is protected by Federal and State Law. Federal regulations prohibit further distribution or copying of this information without permission. If you received this e-mail transmission in error, please notify the sender immediately to arrange for return or destruction of this information. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Helicobacter pylori
We use an Acridine Orange stain. You will need a fluorescent scope for it, though. It's a very simple procedure. Depariffinize slides, air dry, apply AO solution, let stand for 7 minutes, rinse, dry, coverslip from xylene. -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu]on Behalf Of Kathy M. Gorham Sent: Wednesday, October 06, 2010 10:20 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Helicobacter pylori Good Morning Histo land. I would like to know what others are using for a special stain for Helicobacter pylori not IHC. Do you use a kit? From where? Make up your own? Procedure. Thanks you. You have always been so helpful. Kathy Gorham H.T. GRH National Recognition Outstanding Rural Health Organization of 2009 awarded by NRHA Gold Standard Critical Access Hospital 2009 awarded by LarsonAllen LLP Leader in Innovative Excellence 2009 awarded by the OAHHS Financial Excellence Award 2010 awarded by the national Rural Health Research Policy Analysis Center Healthcare Achievement Award for Quality in Patient Care Delivery and Satisfaction 2010 awarded by Amerinet GRH Mission We will ensure access to high-quality, cost-effective health services in a safe, customer-friendly environment for all those in need of our services. GRH Confidentiality Notice This e-mail and any attached documents are for the intended recipient/s only and should be protected against viewing by unauthorized persons. The information herein may have been disclosed from records whose confidentiality is protected by Federal and State Law. Federal regulations prohibit further distribution or copying of this information without permission. If you received this e-mail transmission in error, please notify the sender immediately to arrange for return or destruction of this information. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet CONFIDENTIALITY NOTICE This message and any included attachments are from Somerset Medical Center and are intended only for the addressee. The information contained in this message is confidential and may contain privileged, confidential, proprietary and/or trade secret information entitled to protection and/or exemption from disclosure under applicable law. Unauthorized forwarding, printing, copying, distribution, or use of such information is strictly prohibited and may be unlawful. If you are not the addressee, please promptly delete this message and notify the sender of the delivery error by e-mail or you may call Somerset Medical Center's computer Help Desk at 908-685-2200, ext. 4050. Be sure to visit Somerset Medical Center's Web site - www.somersetmedicalcenter.com - for the most up-to-date news, event listings, health information and more. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] AChE fiber staining protocoll?
IdĂŠzet (Houston, Ronald ronald.hous...@nationwidechildrens.org): Thank you for the answer. We will try this protocol and i will reply the results. Csaba Szigeti I have had great success in fine fibers with the modification of Martucciello et al, Eur J Pediatr Surg 2001; 11: 300-304, but only in 15 micron intestine sections, so I do not know how it will transpose to brain studies and the thicker sections Ronnie Houston Anatomic Pathology Manager Nationwide Children's Hospital Columbus OH 43205 (614) 722 5450 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of szi...@bio.u-szeged.hu Sent: Wednesday, October 06, 2010 3:34 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] AChE fiber staining protocoll? Dear Histonet Members! We are going to investigate AChE fiber density changes in the cortex of transcardially perfused rat brain slices (30 micrometer, criostat sections, perfusing solution: 4% formaldehyde in PB ph 7.4). We need a protocoll to visualize FINE fiber structure to be able to count changes. We used modification of Hedreen and original Tago protocoll. Hedreen good results for general, but no fiber staining, Tago no sucsses. Could you please tell us some advise, protocolls functioning on teh above mentioned objects? Thank you all in advance... Csaba Szigeti This message was sent using IMP, the Internet Messaging Program. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet - Confidentiality Notice: The following mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. The recipient is responsible to maintain the confidentiality of this information and to use the information only for authorized purposes. If you are not the intended recipient (or authorized to receive information for the intended recipient), you are hereby notified that any review, use, disclosure, distribution, copying, printing, or action taken in reliance on the contents of this e-mail is strictly prohibited. If you have received this communication in error, please notify us immediately by reply e-mail and destroy all copies of the original message. Thank you. This message was sent using IMP, the Internet Messaging Program. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: Helicobacter pylori
We use what we call a modified Genta. That is a modified Steiner if done manually. Because we have the Artisan and DAKO didn't have a Genta kit, I tried using the Warthin Starry and adding the Alcian blue and HE manually and it worked fine. So we have not renamed it a modified Warthin Starry! But the H. pylori are very visible as well as the mucin and the morphology. J Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 678-843-7376 - Phone 678-843-7831 - Fax -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Kathy M. Gorham Sent: Wednesday, October 06, 2010 10:20 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Helicobacter pylori Good Morning Histo land. I would like to know what others are using for a special stain for Helicobacter pylori not IHC. Do you use a kit? From where? Make up your own? Procedure. Thanks you. You have always been so helpful. Kathy Gorham H.T. GRH National Recognition Outstanding Rural Health Organization of 2009 awarded by NRHA Gold Standard Critical Access Hospital 2009 awarded by LarsonAllen LLP Leader in Innovative Excellence 2009 awarded by the OAHHS Financial Excellence Award 2010 awarded by the national Rural Health Research Policy Analysis Center Healthcare Achievement Award for Quality in Patient Care Delivery and Satisfaction 2010 awarded by Amerinet GRH Mission We will ensure access to high-quality, cost-effective health services in a safe, customer-friendly environment for all those in need of our services. GRH Confidentiality Notice This e-mail and any attached documents are for the intended recipient/s only and should be protected against viewing by unauthorized persons. The information herein may have been disclosed from records whose confidentiality is protected by Federal and State Law. Federal regulations prohibit further distribution or copying of this information without permission. If you received this e-mail transmission in error, please notify the sender immediately to arrange for return or destruction of this information. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice: This e-mail, including any attachments is the property of Catholic Health East and is intended for the sole use of the intended recipient(s). It may contain information that is privileged and confidential. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please delete this message, and reply to the sender regarding the error in a separate email. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] PAS STAIN
Hi We have been doing a PAS stain on the same control block and same reagent supplier for a long time. Yesterday when we ran the slides, they failed to stain We opened new bottles of Periodic Acid and Schiff's Reagent and recut fresh controls Still, we are unable to get any staining. Suggestions to help would be appreciated Diana ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Histokinette 2000
I have a Leica Histokinette 2000 that needs one beaker too be usable. Does anyone know where I can buy one of these? IMEB, inc does not have them. Jo Ann Biedermann Research Assistant University of Arkansas for Medical Sciences Reynolds Institute on Aging 629 Jack Stephens Drive Room 3173Mail Slot 807 Little Rock, AR 72205 Phone: 501-526-5803 FAX: 501-526-5830 jabiederm...@uams.edumailto:jabiederm...@uams.edu Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message.. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] PAS STAIN
Were the new bottles from the same lot? -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu]on Behalf Of Diana McCaig Sent: Wednesday, October 06, 2010 11:03 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] PAS STAIN Hi We have been doing a PAS stain on the same control block and same reagent supplier for a long time. Yesterday when we ran the slides, they failed to stain We opened new bottles of Periodic Acid and Schiff's Reagent and recut fresh controls Still, we are unable to get any staining. Suggestions to help would be appreciated Diana ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet CONFIDENTIALITY NOTICE This message and any included attachments are from Somerset Medical Center and are intended only for the addressee. The information contained in this message is confidential and may contain privileged, confidential, proprietary and/or trade secret information entitled to protection and/or exemption from disclosure under applicable law. Unauthorized forwarding, printing, copying, distribution, or use of such information is strictly prohibited and may be unlawful. If you are not the addressee, please promptly delete this message and notify the sender of the delivery error by e-mail or you may call Somerset Medical Center's computer Help Desk at 908-685-2200, ext. 4050. Be sure to visit Somerset Medical Center's Web site - www.somersetmedicalcenter.com - for the most up-to-date news, event listings, health information and more. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Query Special Stainers
Hi All, we have an old Ventana benchmark NexES 9.0 that we used to use for our IHC. We now have a different IHC platform that we use and would like to change the Ventana over to use for running our special stains. Does anyone use the Ventana for their specials and have any advice or comments about how it performs? Thanks! Joanne Clark, HT Histology Supervisor Pathology Consultants of New Mexico ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Helicobacter pylori
Hello, We use what is pretty much a diff quik stain, the stain is called QW and it is from poly scientific. The procedure is really easy. Deparaffinize, alcohols, methanol, water and then we do 15 slow dips in the QW2, 15 slow dips in the QW3, wash in water, 2 quick dips in alcohol and then dry in the oven, coverslip. Depending on your pathologist's preference you may need to adjust the dips in QW3 or the dips in the alcohol (which will decolorize slightly). Hope this helps! Brandi Higgins, BS, HT(ASCP) On Wed, Oct 6, 2010 at 10:19 AM, Kathy M. Gorham km...@grh.org wrote: Good Morning Histo land. I would like to know what others are using for a special stain for Helicobacter pylori not IHC. Do you use a kit? From where? Make up your own? Procedure. Thanks you. You have always been so helpful. Kathy Gorham H.T. GRH National Recognition Outstanding Rural Health Organization of 2009 awarded by NRHA Gold Standard Critical Access Hospital 2009 awarded by LarsonAllen LLP Leader in Innovative Excellence 2009 awarded by the OAHHS Financial Excellence Award 2010 awarded by the national Rural Health Research Policy Analysis Center Healthcare Achievement Award for Quality in Patient Care Delivery and Satisfaction 2010 awarded by Amerinet GRH Mission We will ensure access to high-quality, cost-effective health services in a safe, customer-friendly environment for all those in need of our services. GRH Confidentiality Notice This e-mail and any attached documents are for the intended recipient/s only and should be protected against viewing by unauthorized persons. The information herein may have been disclosed from records whose confidentiality is protected by Federal and State Law. Federal regulations prohibit further distribution or copying of this information without permission. If you received this e-mail transmission in error, please notify the sender immediately to arrange for return or destruction of this information. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
AW: [Histonet] PAS STAIN
You can test the activity of the Schiffs. Add a few drops of formalin to a small amount of Schiffs. There should be the typical pink stain. The pH should be about 2. If there are white flakes in the bottle, this could be a sign for a too high pH. Is your periodic acid ok? Gudrun -Ursprüngliche Nachricht- Von: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von Diana McCaig Gesendet: Mittwoch, 06. Oktober 2010 17:03 An: histonet@lists.utsouthwestern.edu Betreff: [Histonet] PAS STAIN Hi We have been doing a PAS stain on the same control block and same reagent supplier for a long time. Yesterday when we ran the slides, they failed to stain We opened new bottles of Periodic Acid and Schiff's Reagent and recut fresh controls Still, we are unable to get any staining. Suggestions to help would be appreciated Diana ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Where to buy Michels RE: [Histonet] DIF Transport Media
Dear Tony, Thank you for the fine reply on Michel's Transport Media. We had excellent results with human renal biopsies destined for immunofluorescence staining. I don't recall ever exceeding 48 hours in Michels as 72 hours was allowed per recommendation from the original and Elias's publications. The kidney morphology didn't suffer excessively from this transport media as seen on our HE stained frozen sections from Liquid Nitrogen cooled isopentane snap frozen needle biopsies. My renal pathologist always commented that the frozen section HE looked very much like his the FFPE HE section. This must vary from laboratory to laboratory and also what tissue was transported e.g. skin biopsies versus kidney biopsies. We always were very careful with good Michels buffer rinses. As for where to access Michels Transport Media and Michels buffer, we purchased these from Poly Scientific in much larger volumes at a cheaper price than Wampole(?)(Zeus). Aliquots were made and distributed to laboratories although that may not be ideal since busy laboratories may find this a bit labor intensive. I know of laboratories who make up Michels with great success. As for transporting tissue on saline soaked gauze, I can't stress the importance enough to NEVER let the tissue dry out, and snap freeze it as soon as it arrives in the laboratory. I would prefer receiving a tissue in cell culture fluid to ensure no drying. The reference is much appreciated. G'day and missed seeing you at NSH symposium/convention this year. Gayle M. Callis HTL/HT/MT(ASCP) -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Tony Henwood Sent: Tuesday, October 05, 2010 5:44 PM To: Della Speranza, Vinnie; kristen arvidson; histonet Subject: RE: [Histonet] DIF Transport Media Vinnie, I hope you are well. The following might be of use: Specimens for immunofluorescence are usually submitted to the laboratory in cell culture fluid (eg Hanks or RPMI) or on saline soaked gauze. For transport to other institutions, Michel's Transport Medium has often been used: Michel's Buffer 1M potassium citrate, pH 7.02.5 ml 0.1M magnesium sulphate 5.0 ml 0.1M N ethylmalemide5.0 ml Distilled H2O 87.5 ml * Mix well and store at 2 8oC. Exp. 1 year Michel's Transport Medium Michel's Buffer 100ml Ammonium sulphate 55gm Adjust pH to 7.0 7.4 with 1M KOH. Store at 2 8oC. Exp. 1 year Unfortunately Michel's Transport Medium has erroneously been called Michel's Fixative. None of the components of Michel's Transport Medium is a fixative. Ammonium sulphate precipitates antigen-antibody complexes in diseased skin and renal tissues. N ethylmalemide modifies free sulphydryl groups of cysteine residues in proteins (Fischer 2006). Michel's Transport Medium has been shown to be deleterious to morphology. Ultrastructurally, complete destruction of plasma membranes and intracytoplasmic organelles occurs after 48 hours storage. On the other hand, antigenicity is well preserved even after many days storage (Fischer 2006). Fischer (2006) Intern J Surg Pathol 14(1):108. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) 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-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Della Speranza, Vinnie Sent: Wednesday, 6 October 2010 6:10 AM To: 'kristen arvidson'; histonet Subject: RE: [Histonet] DIF Transport Media Zeus was a company that used to market michel's under their own label. I don't believe zeus still exists. Michel's is the name associated with the author of the original paper. I don't have the reference. This solution does not require refrigeration. Vinnie Della Speranza Manager for Anatomic Pathology Services Medical University of South Carolina 165 Ashley Avenue Suite 309 Charleston, South Carolina 29425 Tel: (843) 792-6353 Fax: (843) 792-8974 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of kristen arvidson Sent: Monday, October 04, 2010 2:30 PM To: histonet Subject: [Histonet] DIF Transport Media Is Michel Medium the same as Zeus? Do these need to be refrigerated? Thanks. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: Query Special Stainers
We have 2 Ventana special stainers and they are very reliable work horses. They run every day over half the day and put out very consistent results. Very user friendly too. Jan M Omaha -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Joanne Clark Sent: Wednesday, October 06, 2010 10:22 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Query Special Stainers Hi All, we have an old Ventana benchmark NexES 9.0 that we used to use for our IHC. We now have a different IHC platform that we use and would like to change the Ventana over to use for running our special stains. Does anyone use the Ventana for their specials and have any advice or comments about how it performs? Thanks! Joanne Clark, HT Histology Supervisor Pathology Consultants of New Mexico ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Sponsored by Catholic Health Initiatives and Immanuel, Alegent Health is faithful to the healing ministry of Jesus Christ, providing high quality care for the body, mind and spirit of every person. The information contained in this communication, including attachments, is confidential and private and intended only for the use of the addressees. Unauthorized use, disclosure, distribution or copying is strictly prohibited and may be unlawful. If you received this communication in error, please inform us of the erroneous delivery by return e-mail message from your computer. Additionally, although all attachments have been scanned at the source for viruses, the recipient should check any attachments for the presence of viruses before opening. Alegent Health accepts no liability for any damage caused by any virus transmitted by this e-mail. Thank you for your cooperation. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] PAS STAIN
I suspect bad Periodic acid. We never buy periodic acid already in solution and if that has been sitting around even in a kit, it may have gone bad. In a workshop taught by Charles Culling, an expert on PAS staining, he stressed making periodic acid fresh daily or each time the stain is done. It is not expensive, goes into solution readily, then toss the periodic acid to never be reused with exception of that one day. Also, you can test your Schiffs reagent by putting a few drops of Schiffs into 10 mls NBF, watch it turn a very bright pink red instantly. If it turns purplish, it is not good. It may be a bad kit, but you never said you were using a kit only new bottles of . Consequently we buy periodic acid in crystalline form, make up 1% Periodic acid, and buy the Schiffs Reagent from Fisher, never a kit. Sigma also sells good Schiffs, but Fisher Scientific aka Thermo has larger quantity for less price. We also store our Schiffs in the refrigerator rather than room temperature, a habit left over from days when we made this reagent up in house. Gayle M. Callis HTL/HT/MT(ASCP) -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Rathborne, Toni Sent: Wednesday, October 06, 2010 9:06 AM To: Diana McCaig; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] PAS STAIN Were the new bottles from the same lot? -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu]on Behalf Of Diana McCaig Sent: Wednesday, October 06, 2010 11:03 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] PAS STAIN Hi We have been doing a PAS stain on the same control block and same reagent supplier for a long time. Yesterday when we ran the slides, they failed to stain We opened new bottles of Periodic Acid and Schiff's Reagent and recut fresh controls Still, we are unable to get any staining. Suggestions to help would be appreciated Diana ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet CONFIDENTIALITY NOTICE This message and any included attachments are from Somerset Medical Center and are intended only for the addressee. The information contained in this message is confidential and may contain privileged, confidential, proprietary and/or trade secret information entitled to protection and/or exemption from disclosure under applicable law. Unauthorized forwarding, printing, copying, distribution, or use of such information is strictly prohibited and may be unlawful. If you are not the addressee, please promptly delete this message and notify the sender of the delivery error by e-mail or you may call Somerset Medical Center's computer Help Desk at 908-685-2200, ext. 4050. Be sure to visit Somerset Medical Center's Web site - www.somersetmedicalcenter.com - for the most up-to-date news, event listings, health information and more. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet __ Information from ESET Smart Security, version of virus signature database 5506 (20101005) __ The message was checked by ESET Smart Security. http://www.eset.com __ Information from ESET Smart Security, version of virus signature database 5506 (20101005) __ The message was checked by ESET Smart Security. http://www.eset.com __ Information from ESET Smart Security, version of virus signature database 5510 (20101006) __ The message was checked by ESET Smart Security. http://www.eset.com __ Information from ESET Smart Security, version of virus signature database 5510 (20101006) __ The message was checked by ESET Smart Security. http://www.eset.com ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: Query Special Stainers
We also have 2 Vantnan Special stainers and totally agree. Marcia M mason City Marcia Funk Histology Laboratory Mercy Medical Center North Iowa Mason City, IA, 50401 641-422-7907 Mahoney,Janice A janice.maho...@alegent.org 10/06/2010 10:49 AM We have 2 Ventana special stainers and they are very reliable work horses. They run every day over half the day and put out very consistent results. Very user friendly too. Jan M Omaha -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Joanne Clark Sent: Wednesday, October 06, 2010 10:22 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Query Special Stainers Hi All, we have an old Ventana benchmark NexES 9.0 that we used to use for our IHC. We now have a different IHC platform that we use and would like to change the Ventana over to use for running our special stains. Does anyone use the Ventana for their specials and have any advice or comments about how it performs? Thanks! Joanne Clark, HT Histology Supervisor Pathology Consultants of New Mexico ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Sponsored by Catholic Health Initiatives and Immanuel, Alegent Health is faithful to the healing ministry of Jesus Christ, providing high quality care for the body, mind and spirit of every person. The information contained in this communication, including attachments, is confidential and private and intended only for the use of the addressees. Unauthorized use, disclosure, distribution or copying is strictly prohibited and may be unlawful. If you received this communication in error, please inform us of the erroneous delivery by return e-mail message from your computer. Additionally, although all attachments have been scanned at the source for viruses, the recipient should check any attachments for the presence of viruses before opening. Alegent Health accepts no liability for any damage caused by any virus transmitted by this e-mail. Thank you for your cooperation. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] PAS staining
I have for some time had a problem with Schiff's reagent and PAS staining. - I have tested each new, unopened bottle of Schiff's reagent with formaldehyde and always the color development was immediate, but purple, definately not pink.This result has been quite consistant. - PAS staining for glycogen using the method in *Histotechnology a self Instructional text,* would fail to demonstrate any glycogen in autopsy liver specimens. I teach Histology at a community college and this problem has driven me crazy for a number of years. I have tried several brands of Schiff with the same results. Recently I obtained sheep tissues which were promptly refrigerated and fixed after death, and I had hoped these tissues would demonstrate glycogen. ( My thinking was that perhaps delay before autopsy was somehow diminishing the glycogen in the specimens that I had.or that perhaps long term NBF fixation had hampered staining.) Basement membranes were stained with the Schiff reagent as expected despite the purple color in the formaldehyde test. Hotchkiss Mcmanus with the same reagent also produced beautiful staining of fungus a lovely magenta color. A search of the web made me suspiciious when I noted that Schiff added to 37-40% formaldehyde should produce a pink or red color, however, A spot check of formalin using Schiff should produce a purple color. I sent an e-mail to Brian Hewlett to ask if he could make any recommendation. Brian was not surprised by the purple color devopment in testing, He suggested that a large number of available aldehyde groups would be expected to produce this color. He suggested that I increase my periodate oxidation to 20-30 minutes and my Schiff application to 30 minutes. This worked and I am extremely grateful!. Has anyone else had an experience like this? Janet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] PAS staining
Yes, I have experienced the same thing. Many people forget about the impact temperature has on some staining reactions. If you keep your Schiffs and Periodic acid in the refrigerator it may take longer to stain than it would if the reagents were at room temp. Many old procedures are written with the reagents at room temp (even the ones requiring refrigeration). This was one of those live and learn situations for me. Now temp and time are the first things I look at when a stain does not work on a known control. Jan M Omaha -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Janet Keeping Sent: Wednesday, October 06, 2010 11:43 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] PAS staining I have for some time had a problem with Schiff's reagent and PAS staining. - I have tested each new, unopened bottle of Schiff's reagent with formaldehyde and always the color development was immediate, but purple, definately not pink.This result has been quite consistant. - PAS staining for glycogen using the method in *Histotechnology a self Instructional text,* would fail to demonstrate any glycogen in autopsy liver specimens. I teach Histology at a community college and this problem has driven me crazy for a number of years. I have tried several brands of Schiff with the same results. Recently I obtained sheep tissues which were promptly refrigerated and fixed after death, and I had hoped these tissues would demonstrate glycogen. ( My thinking was that perhaps delay before autopsy was somehow diminishing the glycogen in the specimens that I had.or that perhaps long term NBF fixation had hampered staining.) Basement membranes were stained with the Schiff reagent as expected despite the purple color in the formaldehyde test. Hotchkiss Mcmanus with the same reagent also produced beautiful staining of fungus a lovely magenta color. A search of the web made me suspiciious when I noted that Schiff added to 37-40% formaldehyde should produce a pink or red color, however, A spot check of formalin using Schiff should produce a purple color. I sent an e-mail to Brian Hewlett to ask if he could make any recommendation. Brian was not surprised by the purple color devopment in testing, He suggested that a large number of available aldehyde groups would be expected to produce this color. He suggested that I increase my periodate oxidation to 20-30 minutes and my Schiff application to 30 minutes. This worked and I am extremely grateful!. Has anyone else had an experience like this? Janet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Sponsored by Catholic Health Initiatives and Immanuel, Alegent Health is faithful to the healing ministry of Jesus Christ, providing high quality care for the body, mind and spirit of every person. The information contained in this communication, including attachments, is confidential and private and intended only for the use of the addressees. Unauthorized use, disclosure, distribution or copying is strictly prohibited and may be unlawful. If you received this communication in error, please inform us of the erroneous delivery by return e-mail message from your computer. Additionally, although all attachments have been scanned at the source for viruses, the recipient should check any attachments for the presence of viruses before opening. Alegent Health accepts no liability for any damage caused by any virus transmitted by this e-mail. Thank you for your cooperation. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] PAS staining
Although I have allowed Schiff's and Periodic acid to room temperature, temperature may indeed be a factor. The ventilation system within this lab is extremely effficient and takes in a great deal of outside air. I am in Newfoundland, and lower outside temperatures are more common than we would like. :( Especially during the winter of course.) I just turned on the ventilation and the temperature dropped 2 degrees C in about five minutes. I will watch my room temperature more closely. Janet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] PAS staining
I was taught to avoid aqueous fixatives when trying to retain glycogen, soluble in NBF. Carnoys, Gendres fluid or acid alcohol formalin are three recommendations (Sheehan and Hrapchak Theory and Practice of Histotechnology) followed by starting processing in 95 to 100% alcohols. Alcoholic formalin should also work. It could very well be your long term storage in NBF has removed the glycogen, although basement membranes or fungus would not be affected. This was very apparent in a study done here where they wanted to see glycogen storage in mouse livers fixed in NBF for over a week and routinely processed starting in 70% alcohol. The glycogen, for all purposes, was removed even in experimental animals who had large quantity of glycogen in the cells (faintly stained but not what expected). Certainly increasing time in periodic acid and Schiffs can help. Also, one can increase the percentage of periodic acid from 0.5% to 1%, a hint Culling gave, as long as this is freshly made. However, we never use periodic acid for fungus staining, only chromic acid since periodic acid oxidation can give false negative results with Schiffs reagent. This is published in J Histotechnology by Carson and Fredenburgh. Interesting, but I still get a bright red pink color with Neutral buffered formalin test. I have never used concentrated 37% to 40% stock formaldehyde, only neutral buffered formalin (fixative) which would have fewer aldehyde groups available. Outdated, bad Schiffs always has the obvious purplish color with NBF. One thing we never allow is return used Schiffs back into stock Schiffs. Stock stain solutions are never contaminated with depleted, used solutions. We date when the Schiffs was used, and not reused within the week, this is discarded. This was particularly important with human renal biopsy work with the renal pathologist recommending disposing of used Schiffs. Our biopsy service did not handle a large number of biopsies in a year and this was a way to ensure consistent PAS staining by using only Schiffs. Successful PAS staining of basement membrane on 1 to 2 um zinc formalin or FFPE sections were never a problem. Gayle M. Callis HTL/HT/MT(ASCP) -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Janet Keeping Sent: Wednesday, October 06, 2010 10:43 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] PAS staining I have for some time had a problem with Schiff's reagent and PAS staining. - I have tested each new, unopened bottle of Schiff's reagent with formaldehyde and always the color development was immediate, but purple, definately not pink.This result has been quite consistant. - PAS staining for glycogen using the method in *Histotechnology a self Instructional text,* would fail to demonstrate any glycogen in autopsy liver specimens. I teach Histology at a community college and this problem has driven me crazy for a number of years. I have tried several brands of Schiff with the same results. Recently I obtained sheep tissues which were promptly refrigerated and fixed after death, and I had hoped these tissues would demonstrate glycogen. ( My thinking was that perhaps delay before autopsy was somehow diminishing the glycogen in the specimens that I had.or that perhaps long term NBF fixation had hampered staining.) Basement membranes were stained with the Schiff reagent as expected despite the purple color in the formaldehyde test. Hotchkiss Mcmanus with the same reagent also produced beautiful staining of fungus a lovely magenta color. A search of the web made me suspiciious when I noted that Schiff added to 37-40% formaldehyde should produce a pink or red color, however, A spot check of formalin using Schiff should produce a purple color. I sent an e-mail to Brian Hewlett to ask if he could make any recommendation. Brian was not surprised by the purple color devopment in testing, He suggested that a large number of available aldehyde groups would be expected to produce this color. He suggested that I increase my periodate oxidation to 20-30 minutes and my Schiff application to 30 minutes. This worked and I am extremely grateful!. Has anyone else had an experience like this? Janet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet __ Information from ESET Smart Security, version of virus signature database 5510 (20101006) __ The message was checked by ESET Smart Security. http://www.eset.com __ Information from ESET Smart Security, version of virus signature database 5510 (20101006) __ The message was checked by ESET Smart Security. http://www.eset.com __ Information from ESET Smart Security, version of virus signature database 5510 (20101006) __ The message was checked by ESET Smart
RE: [Histonet] PAS staining
Jan is correct, and unless specified in a staining protocol, room temperature is used. -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Mahoney,Janice A Sent: Wednesday, October 06, 2010 11:20 AM To: 'Janet Keeping'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] PAS staining Yes, I have experienced the same thing. Many people forget about the impact temperature has on some staining reactions. If you keep your Schiffs and Periodic acid in the refrigerator it may take longer to stain than it would if the reagents were at room temp. Many old procedures are written with the reagents at room temp (even the ones requiring refrigeration). This was one of those live and learn situations for me. Now temp and time are the first things I look at when a stain does not work on a known control. Jan M Omaha -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Janet Keeping Sent: Wednesday, October 06, 2010 11:43 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] PAS staining I have for some time had a problem with Schiff's reagent and PAS staining. - I have tested each new, unopened bottle of Schiff's reagent with formaldehyde and always the color development was immediate, but purple, definately not pink.This result has been quite consistant. - PAS staining for glycogen using the method in *Histotechnology a self Instructional text,* would fail to demonstrate any glycogen in autopsy liver specimens. I teach Histology at a community college and this problem has driven me crazy for a number of years. I have tried several brands of Schiff with the same results. Recently I obtained sheep tissues which were promptly refrigerated and fixed after death, and I had hoped these tissues would demonstrate glycogen. ( My thinking was that perhaps delay before autopsy was somehow diminishing the glycogen in the specimens that I had.or that perhaps long term NBF fixation had hampered staining.) Basement membranes were stained with the Schiff reagent as expected despite the purple color in the formaldehyde test. Hotchkiss Mcmanus with the same reagent also produced beautiful staining of fungus a lovely magenta color. A search of the web made me suspiciious when I noted that Schiff added to 37-40% formaldehyde should produce a pink or red color, however, A spot check of formalin using Schiff should produce a purple color. I sent an e-mail to Brian Hewlett to ask if he could make any recommendation. Brian was not surprised by the purple color devopment in testing, He suggested that a large number of available aldehyde groups would be expected to produce this color. He suggested that I increase my periodate oxidation to 20-30 minutes and my Schiff application to 30 minutes. This worked and I am extremely grateful!. Has anyone else had an experience like this? Janet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Sponsored by Catholic Health Initiatives and Immanuel, Alegent Health is faithful to the healing ministry of Jesus Christ, providing high quality care for the body, mind and spirit of every person. The information contained in this communication, including attachments, is confidential and private and intended only for the use of the addressees. Unauthorized use, disclosure, distribution or copying is strictly prohibited and may be unlawful. If you received this communication in error, please inform us of the erroneous delivery by return e-mail message from your computer. Additionally, although all attachments have been scanned at the source for viruses, the recipient should check any attachments for the presence of viruses before opening. Alegent Health accepts no liability for any damage caused by any virus transmitted by this e-mail. Thank you for your cooperation. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet __ Information from ESET Smart Security, version of virus signature database 5510 (20101006) __ The message was checked by ESET Smart Security. http://www.eset.com __ Information from ESET Smart Security, version of virus signature database 5510 (20101006) __ The message was checked by ESET Smart Security. http://www.eset.com __ Information from ESET Smart Security, version of virus signature database 5510 (20101006) __ The message was checked by ESET Smart Security. http://www.eset.com ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: RE: [Histonet] PAS staining
/mailman/listinfo/histonet __ Information from ESET Smart Security, version of virus signature database 5510 (20101006) __ The message was checked by ESET Smart Security. http://www.eset.com __ Information from ESET Smart Security, version of virus signature database 5510 (20101006) __ The message was checked by ESET Smart Security. http://www.eset.com __ Information from ESET Smart Security, version of virus signature database 5510 (20101006) __ The message was checked by ESET Smart Security. http://www.eset.com ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] PAS staining
Security, version of virus signature database 5510 (20101006) __ The message was checked by ESET Smart Security. http://www.eset.com __ Information from ESET Smart Security, version of virus signature database 5510 (20101006) __ The message was checked by ESET Smart Security. http://www.eset.com __ Information from ESET Smart Security, version of virus signature database 5510 (20101006) __ The message was checked by ESET Smart Security. http://www.eset.com __ Information from ESET Smart Security, version of virus signature database 5510 (20101006) __ The message was checked by ESET Smart Security. http://www.eset.com ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: RE: [Histonet] PAS staining
/mailman/listinfo/histonet __ Information from ESET Smart Security, version of virus signature database 5510 (20101006) __ The message was checked by ESET Smart Security. http://www.eset.com __ Information from ESET Smart Security, version of virus signature database 5510 (20101006) __ The message was checked by ESET Smart Security. http://www.eset.com __ Information from ESET Smart Security, version of virus signature database 5510 (20101006) __ The message was checked by ESET Smart Security. http://www.eset.com ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] PAS staining
I still make my own Schiff's reagent. It is easy to make and I've never had a problem with it. Keeps well in refrigerator. I do make the periodic acid fresh each time. If anyone wants the recipe, let me know. LuAnn On 10/6/2010 11:43 AM, Janet Keeping wrote: I have for some time had a problem with Schiff's reagent and PAS staining. - I have tested each new, unopened bottle of Schiff's reagent with formaldehyde and always the color development was immediate, but purple, definately not pink.This result has been quite consistant. - PAS staining for glycogen using the method in *Histotechnology a self Instructional text,* would fail to demonstrate any glycogen in autopsy liver specimens. I teach Histology at a community college and this problem has driven me crazy for a number of years. I have tried several brands of Schiff with the same results. Recently I obtained sheep tissues which were promptly refrigerated and fixed after death, and I had hoped these tissues would demonstrate glycogen. ( My thinking was that perhaps delay before autopsy was somehow diminishing the glycogen in the specimens that I had.or that perhaps long term NBF fixation had hampered staining.) Basement membranes were stained with the Schiff reagent as expected despite the purple color in the formaldehyde test. Hotchkiss Mcmanus with the same reagent also produced beautiful staining of fungus a lovely magenta color. A search of the web made me suspiciious when I noted that Schiff added to 37-40% formaldehyde should produce a pink or red color, however, A spot check of formalin using Schiff should produce a purple color. I sent an e-mail to Brian Hewlett to ask if he could make any recommendation. Brian was not surprised by the purple color devopment in testing, He suggested that a large number of available aldehyde groups would be expected to produce this color. He suggested that I increase my periodate oxidation to 20-30 minutes and my Schiff application to 30 minutes. This worked and I am extremely grateful!. Has anyone else had an experience like this? Janet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Question about Oil Red O controls
Komal, I don't know what kind of lab you are in, I'm in a core facility and I do histology on research projects. When I get an ORO this is what I do for a control: I get a piece of tissue like mouse kidney with some fat attached or maybe some muscle with fat and have it snap frozen. I have found that the frozen blocks stay good for a long time at -80ºC and so do the frozen sections on slides. I always cut a bunch of slides and store them and take one out when I have the stain ordered. Andi Andrea Grantham, HT (ASCP) Senior Research Specialist University of Arizona Cell Biology and Anatomy Histology Service Laboratory P.O.Box 245044 Tucson, AZ 85724 algra...@email.arizona.edu Tel: 520.626.4415 Fax: 520.626.2097 happy slicing and dicing and may all your stains work perfectly - Paula Sicurello P Please consider the environment before printing this email. On Oct 6, 2010, at 6:15 AM, Komal Gada wrote: Hello Histonetters, I am trying to find a procedure for using butter and egg yolks as controls for the Oil Red O stain (to show the fat). Does anyone have something they would be able to share with me? Thanks, Komal ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] PAS STAIN
I make my Schiff's from scratch and store it in the refrigerator. I use a minimum amount of charcoal and I heat it in the oven at over 100 C the night before to be sure it is nice and dry. I make periodic acid fresh that day. I make bisulfite rinses fresh that day. Aqueous formalin for 48 hours at room temp. is OK for rat and mouse liver glycogen, I don't know about other species. Slices of liver should be thin. When in doubt formalin+alcohol+acetic acid is an excellent fixative. Geoff Diana McCaig wrote: Hi We have been doing a PAS stain on the same control block and same reagent supplier for a long time. Yesterday when we ran the slides, they failed to stain We opened new bottles of Periodic Acid and Schiff's Reagent and recut fresh controls Still, we are unable to get any staining. Suggestions to help would be appreciated Diana ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- -- ** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583 mcaul...@umdnj.edu ** ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] PAS STAIN
I know there were a ton of posts about this, but I deleted them o= n
accident...Did you test your Schiff's? Put a drop of 10% formalin i n
it...if it turns pink, it's good...if not, throw it out =)
Sarah Goebel= , B.A., HT (ASCP)
Histotechnician
XBiotech US= A Inc.
8201 East Riverside Dr. Bldg 4 Suite 100
A= ustin, Texas 78744
(512)386-5107
Original Message
Subject: Re: [Histonet] PAS STAIN
From: Geoff McAuliffe [1]mcaul...@um= dnj.edu
Date: Wed, October 06, 2010 1:28 pm
To: Diana McCaig [2]dmcc...@ckha.on.= ca, Histonet ((E-mail))
[3]histo...@lists.uts= outhwestern.edu
I make my Schiff's from scratch and store it in the refrigerator.
I use a minimum amount of charcoal and I heat it in the oven at over
100 C the night before to be sure it is nice and dry.
I make periodic acid fresh that day.
I make bisulfite rinses fresh that day.
Aqueous formalin for 48 hours at room temp. is OK for rat and mouse
liver glycogen, I don't know about other species. Slices of liver
should be thin.
When in doubt formalin+alcohol+acetic acid is an excellent fixative.
Geoff
Diana McCaig wrote:
Hi
We have been doing a PAS stain on the same control block and same
reagent supplier for a long time.
Yesterday when we ran the slides, they failed to stain
We opened new bottles of Periodic Acid and Schiff's Reagent and
recut= br fresh controls
Still, we are unable to get any staining.
Suggestions to help would be appreciated
Diana
___
Histonet mailing list
[4]histo...@lists.ut= southwestern.edu
[5]= http://lists.utsouthwestern.edu/mailman/listinfo/histonet
--
--
**
Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583
[6]mcaul...@umdnj.edu
**
___
Histonet mailing list
[7]histo...@lists.utsouth= western.edu
[8]http:= //lists.utsouthwestern.edu/mailman/listinfo/histonet
References
1. 3Dmailto:mcaul...@umdnj.edu;
2. 3Dmailto:dmcc...@ckha.on.ca;
3. 3Dmailto:histonet@lists.utsouthwestern.edu;
4. 3Dmailto:Histonet@lists.utsouthwestern.edu;
5. 3Dhttp://lists.utsouthwestern.edu/mailman/listinfo/histonet;
6. 3Dmailto:mcaul...@umdnj.edu;
7. 3Dmailto:Histonet@lists.utsouthwestern.edu;
8. 3Dhttp://lists.utsouthwestern.edu/mailman/listinfo/histonet;
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[Histonet] IL1a
Has anybody ever worked with IL1-alpha for IHC? Sarah Goebel, B.A., HT (ASCP) Histotechnician XBiotech USA Inc. 8201 East Riverside Dr. Bldg 4 Suite 100 Austin, Texas 78744 (512)386-5107 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] PAS staining
Janet, The major problem I have encounted with the PAS stain is breakdown of the Schiff's reagent (white precipitate). Replacement of the Schiff's reagent usually (?always) solves this. I am not surprised that glycogen in autopsy tissues is difficult to demonstrate. Post-mortem seems to decrease the glycogen levels. I prefer to use bowel and kidney (mucin basement membranes) to initially check the solutions. Interestingly we include glycogen-rich liver in our PAS control block and if you audit the PAS controls from when the Schiff's bottle is opened until it is empty, or when staining decreases, you will notice a gradual loss of glycogen staining. Decrease in PAS staining of Fungi and basement membranes is often not as apparent. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) 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-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Janet Keeping Sent: Thursday, 7 October 2010 3:43 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] PAS staining I have for some time had a problem with Schiff's reagent and PAS staining. - I have tested each new, unopened bottle of Schiff's reagent with formaldehyde and always the color development was immediate, but purple, definately not pink.This result has been quite consistant. - PAS staining for glycogen using the method in *Histotechnology a self Instructional text,* would fail to demonstrate any glycogen in autopsy liver specimens. I teach Histology at a community college and this problem has driven me crazy for a number of years. I have tried several brands of Schiff with the same results. Recently I obtained sheep tissues which were promptly refrigerated and fixed after death, and I had hoped these tissues would demonstrate glycogen. ( My thinking was that perhaps delay before autopsy was somehow diminishing the glycogen in the specimens that I had.or that perhaps long term NBF fixation had hampered staining.) Basement membranes were stained with the Schiff reagent as expected despite the purple color in the formaldehyde test. Hotchkiss Mcmanus with the same reagent also produced beautiful staining of fungus a lovely magenta color. A search of the web made me suspiciious when I noted that Schiff added to 37-40% formaldehyde should produce a pink or red color, however, A spot check of formalin using Schiff should produce a purple color. I sent an e-mail to Brian Hewlett to ask if he could make any recommendation. Brian was not surprised by the purple color devopment in testing, He suggested that a large number of available aldehyde groups would be expected to produce this color. He suggested that I increase my periodate oxidation to 20-30 minutes and my Schiff application to 30 minutes. This worked and I am extremely grateful!. Has anyone else had an experience like this? Janet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet * This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This note also confirms that this email message has been virus scanned and although no computer viruses were detected, The Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. * ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] HMGB
Hi, Sorry about taking so long to get back to you. The antibody I'm using is from abcam, cat# ab18256. I really think the 1:100 or 1:200 is the best dilution so far. Amos On Mon, Oct 4, 2010 at 1:01 PM, histonet-requ...@lists.utsouthwestern.eduwrote: Message: 1 Date: Mon, 4 Oct 2010 08:21:15 -0500 From: Fabrice GANKAM gan...@googlemail.com Subject: Re: [Histonet] HMGB and RAGE; TLR2, TLR4 antibodies To: Amos Brooks amosbro...@gmail.com Cc: histonet@lists.utsouthwestern.edu Message-ID: aanlkti=dfhur43vxj6ypjvs8dfnpcaq9b88kzbrcz...@mail.gmail.com Content-Type: text/plain; charset=ISO-8859-1 thanks Amos which one did you used ? you have catalog number ? ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Question about Oil Red O controls
Believe it or not, mayonnaise makes a great control for Oil Red O. Steve -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Andrea Grantham Sent: Wednesday, October 06, 2010 3:42 PM Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Question about Oil Red O controls Komal, I don't know what kind of lab you are in, I'm in a core facility and I do histology on research projects. When I get an ORO this is what I do for a control: I get a piece of tissue like mouse kidney with some fat attached or maybe some muscle with fat and have it snap frozen. I have found that the frozen blocks stay good for a long time at -80ºC and so do the frozen sections on slides. I always cut a bunch of slides and store them and take one out when I have the stain ordered. Andi Andrea Grantham, HT (ASCP) Senior Research Specialist University of Arizona Cell Biology and Anatomy Histology Service Laboratory P.O.Box 245044 Tucson, AZ 85724 algra...@email.arizona.edu Tel: 520.626.4415 Fax: 520.626.2097 happy slicing and dicing and may all your stains work perfectly - Paula Sicurello P Please consider the environment before printing this email. On Oct 6, 2010, at 6:15 AM, Komal Gada wrote: Hello Histonetters, I am trying to find a procedure for using butter and egg yolks as controls for the Oil Red O stain (to show the fat). Does anyone have something they would be able to share with me? Thanks, Komal ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
