[Histonet] need recommendations on working alpha synulcein IHC for human tissue

[Maria Mejia]

I'm looking for a working alpha synuclein IHC antibody to be used on human 
tissue.  I work on human
brain stem free-floating 60um sections.  I would greatly appreciate any 
suggestions & recommendations
from anyone.

Regards

Maria Mejia
Histology Supervisor
UCSF
Department of Neurology
SF, CA
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[Histonet] RE: IHC proficiency testing

[Sebree Linda A]

Hi Kari,

I'd suggest taking advantage of CAP proficiency testing surveys so as to 
compare your lab's results to others.

Linda A. Sebree

From: histonet-boun...@lists.utsouthwestern.edu 
[histonet-boun...@lists.utsouthwestern.edu] on behalf of Delray Beach Pathology 
Kari Simeone [ksime...@leavittmgt.com]
Sent: Monday, June 23, 2014 2:24 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] IHC proficiency testing

Can anyone share their CLIA proficiency protocols/programs for automated IHC 
staining? Other than the obvious antibody validation, positive and negative 
controls, QA and QC performed, should we enroll in an actual program for our 
IHC? We currently use two Leica Bonds and about 70 antibodies. No Her2 or ER/PR 
or FISH.

Thanks in advance!



K. Simeone

ksime...@leavittmgt.com





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[Histonet] RE: Slides for Thermo Slidemate

[Walter Benton]

Lecia makes a Thermal charged slide that we just tried and it works great. Best 
print I've seen to date and no smearing.
Walter Benton HT(ASCP)QIHC
Histology Supervisor
Chesapeake Urology Associates
806 Landmark Drive, Suite 127
Glen Burnie, MD 21061
443-471-5850 (Direct)
410-768-5961 (Lab)
410-768-5965 (Fax)
Chesapeakeurology.com

Voted a Best Place to Work by
Baltimore and Modern Healthcare
Magazines.

From: histonet-boun...@lists.utsouthwestern.edu 
[histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Laurie Colbert 
[lcolb...@pathmdlabs.com]
Sent: Monday, June 23, 2014 3:37 PM
To: Histonet Post (histonet@lists.utsouthwestern.edu)
Subject: [Histonet] Slides for Thermo Slidemate

What slides are others using (other than Thermo's Superfrost Plus slides) that 
won't jam in the Slidemate and that will produce print that does not smear?

Laurie Colbert, HT (ASCP)
Histology Supervisor
PATH MD
8158 Beverly Blvd.
Los Angeles, CA  90048
(323) 648-3214 direct
(424) 245-7284 main lab

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[Histonet] SV 40

[Helene Degan]

Hi
Wondering where other laboratories performing the SV40 anitbody for IHC are 
obtaining control tissue from, is it purchased from a company if so which one? 
We are currently performing the SV 40  IHC on Ventana Ultra/ XT and we are not 
getting the appropriate  staining, any help would be greatly appreciated.
 
 
Thanks 
 
Helene 
Assistant Supervisor of Histology
Upstate Medical University
 
 
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[Histonet] Slides for Thermo Slidemate

[Laurie Colbert]

What slides are others using (other than Thermo's Superfrost Plus slides) that 
won't jam in the Slidemate and that will produce print that does not smear?

Laurie Colbert, HT (ASCP)
Histology Supervisor
PATH MD
8158 Beverly Blvd.
Los Angeles, CA  90048
(323) 648-3214 direct
(424) 245-7284 main lab

The information contained in this transmission may contain privileged and 
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above. If you are not the intended recipient, you are hereby notified that any 
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strictly prohibited. If you are not the intended recipient, please contact the 
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[Histonet] IHC proficiency testing

[Delray Beach Pathology Kari Simeone]

Can anyone share their CLIA proficiency protocols/programs for automated IHC 
staining? Other than the obvious antibody validation, positive and negative 
controls, QA and QC performed, should we enroll in an actual program for our 
IHC? We currently use two Leica Bonds and about 70 antibodies. No Her2 or ER/PR 
or FISH.

Thanks in advance!



K. Simeone

ksime...@leavittmgt.com





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the sender immediately by return e-mail and delete the original message.


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[Histonet] Part time/on call position available - Atlanta, GA

[Cooper, Deborah S]

Hi,

We are looking for a person who can come in part-time/on call to cut paraffin 
sections only. The person does not need to be a certified histotech, just 
someone who either has cutting experience or is willing to learn and has 
excellent attention to detail. We would need the person to come in a day or two 
after being called (on a sporadic basis) and would average about 4 to 8 hours a 
week. Position is well suited for a retiree.

Thank you,
Deborah


Deborah Cooper
Research Specialist
Center for Neurodegenerative Disease
Emory University
615 Michael St
WRB Rm 515
Atlanta, GA 30322
404-712-8353



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[Histonet] Eosin leaching

[Debra Siena]

Hi Histonet,


Eosin bleaching is usually due to the alcohols that are used after the eosin, 
if you are in a humid area, the alcohols can actually absorb water from the 
humidity in the air. This may mean a change of how often you rotate or change 
the alcohols, possibly more often than you did during the winter months when 
usually less humid.  Also if you use a graded alcohol after the eosin, maybe 
worth looking into going straight to 100%, I usually always recommend at least 
3 changes 100% alcohol for 1 minute and 3 changes of xylene for 1 minute.  If 
the last alcohol in your staining line is pink, it means that there is eosin 
and that means that there is water.  Eosin loves water and loves to come out of 
solution into the one containing the water.  I have even seen pink xylene in 
some staining lines due to water in xylene, however, not enough that it has 
turned cloudy yet but still have a bit of water.  Xylene will tolerate about 
.5% water before becoming too milky or cloudy to use.  Also when you rotate 
your solutions make sure that you dry the containers well before  refilling and 
by graduating the levels of the alcohols and xylenes to make sure that your 
last container completely covers the slides. Lastly, adding a 95% alcohol 
before your eosin, if you don't have one may help to keep excess water out of 
your eosin which will cause the eosin to break down faster, again eosin loves 
water.  Just spouting off the top of my head but hope something helps out.

Debbie Siena
800.442.3573 ext. 229 | www.statlab.com

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[Histonet] RE: Eosin Leaching

[Gregoire, Rhonda (MAFRD)]

Hi Adrienne,

We see eosin leaching in our summer months when it is very humid in our lab. 
The humidity is >70% for most of the summer.  We change out all of our alcohols 
daily to try and prevent it but we sometimes see it even with doing that. 

Rhonda Gregoire, MLT
Supervisor, Clinical Pathology
Veterinary Diagnostic Services 
Manitoba Agriculture, Food and Rural Development 
545 University Crescent 
Winnipeg, MB 
R3T 5S6 
  
phone 204-945-7641 
fax 204-945-7646 
email rhonda.grego...@gov.mb.ca    

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of 
histonet-requ...@lists.utsouthwestern.edu
Sent: June-23-14 12:02 PM
To: histonet@lists.utsouthwestern.edu
Subject: Histonet Digest, Vol 127, Issue 28

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Today's Topics:

   1. RE: Eosin Leaching (Tony Reilly)
   2. Histology Benchmarking Survey (Lesley Bechtold)


--

Message: 1
Date: Mon, 23 Jun 2014 09:22:28 +1000
From: Tony Reilly 
Subject: RE: [Histonet] Eosin Leaching
To: "Rathborne, Toni" ,   "Sanders, Jeanine
(CDC/OID/NCEZID)" , "'AdrienneAnderson'"
, "'histonet@lists.utsouthwestern.edu'"

Message-ID:



Content-Type: text/plain; charset="iso-8859-1"

Hi Adrienne

Also make sure the wash after your bluing agent is higher than the blueing 
solution or alkalinity of the solution will leach out your eosin.

Regards
Tony


Tony Reilly B.App.Sc,  M.Sc
Chief Scientist
Anatomical Pathology
Pathology Queensland PAH
_
Health Services Support Agency| Department of Health

Building 15, Level 1,?
199 Ipswich Road?
WOOLLOONGABBA? Queensland 4102
Ph: 07 3176 2412
Mob: 0402139411
Fax: 07 3176 2930
Email: tony.reil...@health.qld.gov.au | www.health.qld.gov.au ? ? ?





-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Rathborne, Toni
Sent: Friday, 20 June 2014 10:11 PM
To: Sanders, Jeanine (CDC/OID/NCEZID); 'Adrienne Anderson'; 
'histo...@listsutsouthwestern.edu'
Subject: RE: [Histonet] Eosin Leaching

Is your alcohol level as high as or higher than the level of eosin in the 
containers? If there is residual eosin left at the upper portion of the slide, 
that could be causing the problem.

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Sanders, 
Jeanine (CDC/OID/NCEZID)
Sent: Thursday, June 19, 2014 5:26 PM
To: 'Adrienne Anderson'; 'histonet@lists.utsouthwestern.edu'
Subject: RE: [Histonet] Eosin Leaching

Have you changed your coverslipping mountant?

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Adrienne 
Anderson
Sent: Thursday, June 19, 2014 5:24 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Eosin Leaching

Hello all,

We're having a problem with our eosin bleeding out of the sections. I've read 
that this can be caused by not dehydrating adequately after eosin, but we're 
still having an issue when using fresh alcohols. Does anyone have any other 
ideas as to the cause of this problem?

Thanks in advance for your help!

Adrienne



Adrienne Anderson, BS, HTL(ASCP)
Histotechnologist
Phylogeny, Inc. 
1476 Manning Pkwy, Powell, Ohio 43605
Phone: (614) 846-6161 
Fax:  (877) 591-1815

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[Histonet] ISH preparation

[Jennifer.Johnson]

Hi Folks!
I was hoping to get a little input about preparing samples for ISH.
We currently RNAse-away our tools at the time of collection, toss the tissues 
into formalin, process normally and then cut them using RNAse away cleaned 
microtomes/forceps on a bath of DEPC treated water (in a cleaned water bath).

How do other labs prepare paraffin embedded samples?

Do you use special reagents - formalin?  Do you clean the processor and use 
treated reagents there too?  Do you wash the slides in DEPC treated water?

I am trying to figure out how far we need to go here.  We are a high throughput 
lab and we deal with all kinds of tissues and I don't want to have to take down 
a processor and keep it RNAse free if I don't have to!   I will run the 
experiments and test different methods, but I was hoping some of you could 
share your wisdom and give me a place to start.
Thanks,
JJ

Jennifer Johnson
Staff Scientist
Genzyme Corp.
Department of Pathology
5 Mountain Road
Framingham, MA 01701-9322
Phn - 508-271-3610
Fax - 508-872-9080


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[Histonet] RE: Eosin Leaching

[Marcum, Pamela A]

Have you changed your reagent alcohol or has the company you buy from changed 
where it purchases the reagent alcohol?  Reagent alcohols with higher volumes 
of isopropanol than  5% can cause the eosin to leach out.  It will leave a 
green powder in the bottom of you staining dish.  Methanol and Isopropanol 
should be 5% each with 90% ethanol for the safest blend and what most of us old 
folks always thought reagent alcohol would stay.  We were wrong and different 
companied can change vendors or sources and get something they were not 
expecting too.   If you look up reagent alcohol or SDA reagent alcohol you find 
it has many formulas and the only real requirement by the ATF is to make the 
alcohol "non-drinkable or poison to humans".  I have seen formulas with 5 to 8% 
methanol and isopropanol with as low as 87% ethanol.  The percentage will 
depend on price as they have no idea at the manufacturing level what we use ti 
for or why.

Pam Marcum
UAMS

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Amos, Lori (VDH)
Sent: Monday, June 23, 2014 12:29 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] RE: Eosin Leaching

I too have been experiencing Eosin leaching. I have tried all different 
options: Adding acetic acid to the pre-made Eosin, adding time to the alcohols 
and xylenes that follow, even added a running water wash step after the Eosin ( 
to remove excess). Before this, I have had no problems and had made no changes 
to my daily activities. I rotate solutions daily. Aside from changing the 
company I buy my Eosin from, I am at a loss.
Thanks in advance for your help.
Lori

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of 
histonet-requ...@lists.utsouthwestern.edu
Sent: Monday, June 23, 2014 1:08 PM
To: histonet@lists.utsouthwestern.edu
Subject: Histonet Digest, Vol 127, Issue 28

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When replying, please edit your Subject line so it is more specific than "Re: 
Contents of Histonet digest..."


Today's Topics:

   1. RE: Eosin Leaching (Tony Reilly)
   2. Histology Benchmarking Survey (Lesley Bechtold)


--

Message: 1
Date: Mon, 23 Jun 2014 09:22:28 +1000
From: Tony Reilly 
Subject: RE: [Histonet] Eosin Leaching
To: "Rathborne, Toni" ,   "Sanders, Jeanine
(CDC/OID/NCEZID)" , "'AdrienneAnderson'"
, "'histonet@lists.utsouthwestern.edu'"

Message-ID:



Content-Type: text/plain; charset="iso-8859-1"

Hi Adrienne

Also make sure the wash after your bluing agent is higher than the blueing 
solution or alkalinity of the solution will leach out your eosin.

Regards
Tony


Tony Reilly B.App.Sc,  M.Sc
Chief Scientist
Anatomical Pathology
Pathology Queensland PAH
_
Health Services Support Agency| Department of Health

Building 15, Level 1,?
199 Ipswich Road?
WOOLLOONGABBA? Queensland 4102
Ph: 07 3176 2412
Mob: 0402139411
Fax: 07 3176 2930
Email: tony.reil...@health.qld.gov.au | www.health.qld.gov.au ? ? ?





-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Rathborne, Toni
Sent: Friday, 20 June 2014 10:11 PM
To: Sanders, Jeanine (CDC/OID/NCEZID); 'Adrienne Anderson'; 
'histo...@listsutsouthwestern.edu'
Subject: RE: [Histonet] Eosin Leaching

Is your alcohol level as high as or higher than the level of eosin in the 
containers? If there is residual eosin left at the upper portion of the slide, 
that could be causing the problem.

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Sanders, 
Jeanine (CDC/OID/NCEZID)
Sent: Thursday, June 19, 2014 5:26 PM
To: 'Adrienne Anderson'; 'histonet@lists.utsouthwestern.edu'
Subject: RE: [Histonet] Eosin Leaching

Have you changed your coverslipping mountant?

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Adrienne 
Anderson
Sent: Thursday, June 19, 2014 5:24 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Eosin Leaching

Hello all,

We're having a problem with our eosin bleeding out of the sections. I've read 
that this can be caused by not dehydrating adequately after eosin, but we're 
still having an issue when using fresh alcohol

[Histonet] RE: Eosin Leaching

[Amos, Lori (VDH)]

I too have been experiencing Eosin leaching. I have tried all different 
options: Adding acetic acid to the pre-made Eosin, adding time to the alcohols 
and xylenes that follow, even added a running water wash step after the Eosin ( 
to remove excess). Before this, I have had no problems and had made no changes 
to my daily activities. I rotate solutions daily. Aside from changing the 
company I buy my Eosin from, I am at a loss.
Thanks in advance for your help.
Lori

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of 
histonet-requ...@lists.utsouthwestern.edu
Sent: Monday, June 23, 2014 1:08 PM
To: histonet@lists.utsouthwestern.edu
Subject: Histonet Digest, Vol 127, Issue 28

Send Histonet mailing list submissions to
histonet@lists.utsouthwestern.edu

To subscribe or unsubscribe via the World Wide Web, visit
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Contents of Histonet digest..."


Today's Topics:

   1. RE: Eosin Leaching (Tony Reilly)
   2. Histology Benchmarking Survey (Lesley Bechtold)


--

Message: 1
Date: Mon, 23 Jun 2014 09:22:28 +1000
From: Tony Reilly 
Subject: RE: [Histonet] Eosin Leaching
To: "Rathborne, Toni" ,   "Sanders, Jeanine
(CDC/OID/NCEZID)" , "'AdrienneAnderson'"
, "'histonet@lists.utsouthwestern.edu'"

Message-ID:



Content-Type: text/plain; charset="iso-8859-1"

Hi Adrienne

Also make sure the wash after your bluing agent is higher than the blueing 
solution or alkalinity of the solution will leach out your eosin.

Regards
Tony


Tony Reilly B.App.Sc,  M.Sc
Chief Scientist
Anatomical Pathology
Pathology Queensland PAH
_
Health Services Support Agency| Department of Health

Building 15, Level 1,?
199 Ipswich Road?
WOOLLOONGABBA? Queensland 4102
Ph: 07 3176 2412
Mob: 0402139411
Fax: 07 3176 2930
Email: tony.reil...@health.qld.gov.au | www.health.qld.gov.au ? ? ?





-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Rathborne, Toni
Sent: Friday, 20 June 2014 10:11 PM
To: Sanders, Jeanine (CDC/OID/NCEZID); 'Adrienne Anderson'; 
'histo...@listsutsouthwestern.edu'
Subject: RE: [Histonet] Eosin Leaching

Is your alcohol level as high as or higher than the level of eosin in the 
containers? If there is residual eosin left at the upper portion of the slide, 
that could be causing the problem.

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Sanders, 
Jeanine (CDC/OID/NCEZID)
Sent: Thursday, June 19, 2014 5:26 PM
To: 'Adrienne Anderson'; 'histonet@lists.utsouthwestern.edu'
Subject: RE: [Histonet] Eosin Leaching

Have you changed your coverslipping mountant?

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Adrienne 
Anderson
Sent: Thursday, June 19, 2014 5:24 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Eosin Leaching

Hello all,

We're having a problem with our eosin bleeding out of the sections. I've read 
that this can be caused by not dehydrating adequately after eosin, but we're 
still having an issue when using fresh alcohols. Does anyone have any other 
ideas as to the cause of this problem?

Thanks in advance for your help!

Adrienne



Adrienne Anderson, BS, HTL(ASCP)
Histotechnologist
Phylogeny, Inc. 
1476 Manning Pkwy, Powell, Ohio 43605
Phone: (614) 846-6161 
Fax:  (877) 591-1815

This message, including any attachments, is confidential and may be privileged 
or may contain health information protected by state and federal law.
Information and opinions expressed in this message and/or attachments are those 
of the author and are not necessarily those of the company. 
If you are not the intended recipient, please notify the sender and delete this 
message from your system. 
Any use of this information by individuals other than the intended recipient is 
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[Histonet] Histology Benchmarking Survey

[Lesley Bechtold]

Dear Histonetters,

We are seeking information from our peers in the Histology world.  To better 
understand what other Histology Labs offer, we have created a short survey that 
we hope you will fill out. The link to the survey is below.

Information is a useful tool when preparing budgets, hiring staff, buying 
equipment or making the decision to add new processes to your lab.  Anyone who 
fills out the survey will receive a copy of the de-identified results.

We thank you in advance for participating.

If surveys annoy you, please delete this email.

Thank you.

Lesley


Lesley S. Bechtold
Senior Manager, Histopathology Sciences
The Jackson Laboratory
600 Main St.
Bar Harbor, ME 04609
207-288-6322 (phone)
207-288-6325 (fax)


https://www.surveymonkey.com/s/HistoSurvey2014


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is intended solely for the addressee(s). If you believe you received this email 
by mistake, please notify the sender by return email as soon as possible.
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