[Histonet] short temp but home for Christmas

[Cheryl]

Hi All-
 
Looking for a happy person and a good cutter, preferrably the same person!  
 
A short day shift temp, M-F from ASAP through the Friday before Christmas.  
Fill up the bank account and be home for the holidays.  Experience with small 
biopsies required.  Full travel including per diems, hotel, flights and car 
rental (or reimbursement for using your own vehicle).  
 
For more info, call my cell and leave a message--I'll call you back across the 
afternoon.  It's one of the nicest little labs in the South!
 
281.883.7704


Cheryl Kerry, HT(ASCP) 
Full Staff Inc. 
Staffing the AP Lab by helping one GREAT Tech at a time.  
281.852.9457 Office
800.756.3309 Phone & Fax 
ad...@fullstaff.org 

Sign up for the FREE newsletter AP News--updates, tricks of the trade and 
current issues for Anatomic Pathology Clinical Labs. Send a 'subscribe' request 
to apn...@fullstaff.org. Please include your name and specialty in the body of 
the email.
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[Histonet] Job Opening

[Martin, Gary]

We are a small pathology in the Sacramento area (Placerville) looking
for a certified histotech.  This is a full time position that offers
competitive compensation and full benefits.  You can forward your
information to Gary Martin at gmar...@marshallmedical.org. 

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Re: [Histonet] chemical disposal

[Steven Mello]

Separate and chemical hazardous waste!

Steven Mello,HT(ASCP)

Sent from my iPad

On Nov 28, 2012, at 4:48 PM, Rene J Buesa  wrote:

> You are right and your "general manager" is wrong (as they usually are when 
> dealing with histology issues!).
> René J.
> 
> From: Cynthia Pyse 
> To: histonet@lists.utsouthwestern.edu 
> Sent: Wednesday, November 28, 2012 3:44 PM
> Subject: [Histonet] chemical disposal
> 
> Quick question for Histoland. I am having a debate about DAB disposal. Our
> general manager ( non lab background) insists that the liquid DAB can go
> into a biological hazardous waste. I disagree, it is a chemical and needs to
> be disposed in the chemical hazardous waste. What is everyone else doing to
> dispose of DAB. We are located in NY, I do have those regs. Thanks in
> advance for any and all help.
> 
> Cindy
> 
> 
> 
> Cindy Pyse, CLT, HT (ASCP)
> 
> Laboratory Manager
> 
> X-Cell Laboratories
> 
> 20 Northpointe Parkway Suite 100
> 
> Amherst, NY 14228
> 
> 716-250-9235 etx. 232
> 
> e-mail cp...@x-celllab.com
> 
> 
> 
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RE: [Histonet] Validation of a new tissue processor

[Morken, Timothy]

I don't think it is necessary to test every single stain or antibody you do to 
validate a new tissue processor. Most of the time the only thing changing is 
the machine, not the chemicals you use. The point of validation is to prove 
that the system gives you the same results as the old instrument. Using a 
representative sample of types of stains and antibodies will give you 
confidence that it works fine and save a huge amount of money and time. 

Think about it from this perspective: We get paraffin slides and blocks from 
consult cases from all over the place, processed on many different machines 
with infinitely variable time and chemical protocols. Yet virtually all the 
cases work fine in our immuno staining system. It is rare to find cases that do 
not. That alone tells you that massive validation is not really necessary.

The point of a validation procedure is to plan one that you feel will give you 
confidence that the new instrument gives you the same results as the old one. 
If you do insist on testing "all" your antibodies (we have over 200 here) then 
tissue arrays or multiple tissue blocks will save a lot of time and money. 

Of course, if you go to a new processor that uses totally different  types of 
chemicals and methods then validation of every antibody may be necessary. Even 
then, however, representative markers of various types may suffice. 

Tim Morken
Department of Pathology
UC San Francisco Medical Center


-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Helen Fedor
Sent: Wednesday, November 28, 2012 1:51 PM
To: 'Rene J Buesa'; Howe, Helena; 'histonet@lists.utsouthwestern.edu'
Subject: RE: [Histonet] Validation of a new tissue processor

You will also need to validate all of the IHC stains and special stains that 
you do on these tissues. You can take samples from each tissue type and run 
them in parallel on both machines. Then test all of your special stains and 
antibodies that you have in your inventory.

:)
Helen

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa
Sent: Wednesday, November 28, 2012 4:47 PM
To: Howe, Helena; 'histonet@lists.utsouthwestern.edu'
Subject: Re: [Histonet] Validation of a new tissue processor

Using at least 25 different types of tissues, prepare parallel almost identical 
pairs of slices for each tissue and process them simultaneously in your new 
tissue processor and in the old one.
Prepare H&E sections and give them to the pathologists concealing the used 
tissue processor. The pathologists should decide if there are differences 
between the pairs of sections.
The pathologist's opinion should be registered in the QC file and that will be 
your validation.
René J.

From: "Howe, Helena" 
To: "'histonet@lists.utsouthwestern.edu'"  
Sent: Wednesday, November 28, 2012 2:57 PM
Subject: [Histonet] Validation of a new tissue processor

Does anyone know what the CAP is requiring to validate a new tissue processor?
Thank You
Helena Howe
ProMedica Laboratories
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[Histonet] microarray on OCT blocks

[Reynolds,Donna M]

I have an investigator wanting to make microarrays from some frozen OCT blocks. 
I seem to remember reading a way to do this. Can someone please point me in the 
right direction?

Thanks
Donna Reynolds, HT (ASCP) Chief IHC Core Lab
Dept. Cancer Biology,
Univ. Tex. M.D. Anderson Cancer Center
Houston, Texas
713-792-8106

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RE: [Histonet] Validation of a new tissue processor

[Helen Fedor]

You will also need to validate all of the IHC stains and special stains that 
you do on these tissues. You can take samples from each tissue type and run 
them in parallel on both machines. Then test all of your special stains and 
antibodies that you have in your inventory.

:)
Helen

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa
Sent: Wednesday, November 28, 2012 4:47 PM
To: Howe, Helena; 'histonet@lists.utsouthwestern.edu'
Subject: Re: [Histonet] Validation of a new tissue processor

Using at least 25 different types of tissues, prepare parallel almost identical 
pairs of slices for each tissue and process them simultaneously in your new 
tissue processor and in the old one.
Prepare H&E sections and give them to the pathologists concealing the used 
tissue processor. The pathologists should decide if there are differences 
between the pairs of sections.
The pathologist's opinion should be registered in the QC file and that will be 
your validation.
René J.

From: "Howe, Helena" 
To: "'histonet@lists.utsouthwestern.edu'"  
Sent: Wednesday, November 28, 2012 2:57 PM
Subject: [Histonet] Validation of a new tissue processor

Does anyone know what the CAP is requiring to validate a new tissue processor?
Thank You
Helena Howe
ProMedica Laboratories
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _

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Re: [Histonet] chemical disposal

[Rene J Buesa]

You are right and your "general manager" is wrong (as they usually are when 
dealing with histology issues!).
René J.

From: Cynthia Pyse 
To: histonet@lists.utsouthwestern.edu 
Sent: Wednesday, November 28, 2012 3:44 PM
Subject: [Histonet] chemical disposal

Quick question for Histoland. I am having a debate about DAB disposal. Our
general manager ( non lab background) insists that the liquid DAB can go
into a biological hazardous waste. I disagree, it is a chemical and needs to
be disposed in the chemical hazardous waste. What is everyone else doing to
dispose of DAB. We are located in NY, I do have those regs. Thanks in
advance for any and all help.

Cindy



Cindy Pyse, CLT, HT (ASCP)

Laboratory Manager

X-Cell Laboratories

20 Northpointe Parkway Suite 100

Amherst, NY 14228

716-250-9235 etx. 232

e-mail cp...@x-celllab.com



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Re: [Histonet] Validation of a new tissue processor

[Rene J Buesa]

Using at least 25 different types of tissues, prepare parallel almost identical 
pairs of slices for each tissue and process them simultaneously in your new 
tissue processor and in the old one.
Prepare H&E sections and give them to the pathologists concealing the used 
tissue processor. The pathologists should decide if there are differences 
between the pairs of sections.
The pathologist's opinion should be registered in the QC file and that will be 
your validation.
René J.

From: "Howe, Helena" 
To: "'histonet@lists.utsouthwestern.edu'"  
Sent: Wednesday, November 28, 2012 2:57 PM
Subject: [Histonet] Validation of a new tissue processor

Does anyone know what the CAP is requiring to validate a new tissue processor?
Thank You
Helena Howe
ProMedica Laboratories
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _

EMAIL CONFIDENTIALITY NOTICE 
This Email message, and any attachments, may contain confidential patient 
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of this information is prohibited from disclosing this information to any other 
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Re: [Histonet] chemical disposal

[Will Chappell]

Chemical hazardous waste. 

Sent from my iPhone

On Nov 28, 2012, at 12:44 PM, "Cynthia Pyse"  wrote:

> Quick question for Histoland. I am having a debate about DAB disposal. Our
> general manager ( non lab background) insists that the liquid DAB can go
> into a biological hazardous waste. I disagree, it is a chemical and needs to
> be disposed in the chemical hazardous waste. What is everyone else doing to
> dispose of DAB. We are located in NY, I do have those regs. Thanks in
> advance for any and all help.
> 
> Cindy
> 
> 
> 
> Cindy Pyse, CLT, HT (ASCP)
> 
> Laboratory Manager
> 
> X-Cell Laboratories
> 
> 20 Northpointe Parkway Suite 100
> 
> Amherst, NY 14228
> 
> 716-250-9235 etx. 232
> 
> e-mail cp...@x-celllab.com
> 
> 
> 
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[Histonet] California HT Position NEED TO FILL ASAP

[Hale, Meredith]

Great full time  opportunity  for a Histotechnician in Yuba City! North Valley 
GI   is looking for certified HT or HTL  to join their  laboratory . HT 
candidate must meet the following criteria:

* Meet CLIA Grossing Requirements : CFR  493.1489,  
http://wwwn.cdc.gov/clia/regs/toc.aspx/ ,prior experience grossing GI specimens

* HT/HTL ASCP Certified , Bachelor's degree or Associates is required 
.Supervisory experience is preferred.
Duties include:

* Grossing

* Embedding

* Microtomy

* Staining

* Ability to be flexible and take on additional duties' as needed


These positions offer a competitive rate of $35.00 an hour and flexible hours.  
Interested applicants should contact Meredith Hale; phone 214-596-2219 or 
through email mh...@miracals.com


Meredith Hale HT  (ASCP)cm
Operations Liaision Director and Education Coordinator

Miraca Life Sciences
6655 North MacArthur Blvd.
Irving , Texas 75039
Office: 214-596-2219
Cell: 469-648-8253
Fax: 1-866-688-3280
mh...@miracals.com>

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RE: [Histonet] chemical disposal

[Weems, Joyce K.]

It should be separate.

Joyce Weems
Pathology Manager
678-843-7376 Phone
678-843-7831 Fax
joyce.we...@emoryhealthcare.org



www.saintjosephsatlanta.org
5665 Peachtree Dunwoody Road
Atlanta, GA 30342

This e-mail, including any attachments is the property of Saint Joseph's 
Hospital 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.


-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Cynthia Pyse
Sent: Wednesday, November 28, 2012 3:45 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] chemical disposal

Quick question for Histoland. I am having a debate about DAB disposal. Our 
general manager ( non lab background) insists that the liquid DAB can go into a 
biological hazardous waste. I disagree, it is a chemical and needs to be 
disposed in the chemical hazardous waste. What is everyone else doing to 
dispose of DAB. We are located in NY, I do have those regs. Thanks in advance 
for any and all help.

Cindy



Cindy Pyse, CLT, HT (ASCP)

Laboratory Manager

X-Cell Laboratories

20 Northpointe Parkway Suite 100

Amherst, NY 14228

716-250-9235 etx. 232

e-mail cp...@x-celllab.com



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[Histonet] chemical disposal

[Cynthia Pyse]

Quick question for Histoland. I am having a debate about DAB disposal. Our
general manager ( non lab background) insists that the liquid DAB can go
into a biological hazardous waste. I disagree, it is a chemical and needs to
be disposed in the chemical hazardous waste. What is everyone else doing to
dispose of DAB. We are located in NY, I do have those regs. Thanks in
advance for any and all help.

Cindy

 

Cindy Pyse, CLT, HT (ASCP)

Laboratory Manager

X-Cell Laboratories

20 Northpointe Parkway Suite 100

Amherst, NY 14228

716-250-9235 etx. 232

e-mail cp...@x-celllab.com

 

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[Histonet] Multiple Histology Mgmt/Supv Openings in NY, NJ, CT

[Brian-Prometheus]

Hope you had a great Thanksgiving!

 

I am currently working on multiple openings in the tri-state area:

 

IHC Supervisor- Suffern NY

 

Lab Operations Manager (over cyto,histo, pre/post technical) Westchester Cty
NY

 

Histology Supervisor w Hospital Hartford CT

 

Histology Supervisor reference lab northern NJ

 

 

 

 

If you might know anyone currently looking would greatly appreciate it!

 

Thanks

 

 

 

 

Brian Feldman
Principal
Prometheus Healthcare
Office 301-693-9057
Fax 301-368-2478
br...@prometheushealthcare.com
www.prometheushealthcare.com  
*** Stay up to date on the newest positions and healthcare trends nationwide
on Twitter!***
 http://twitter.com/PrometheusBlog

 

 

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[Histonet] Validation of a new tissue processor

[Howe, Helena]

Does anyone know what the CAP is requiring to validate a new tissue processor?
Thank You
Helena Howe
ProMedica Laboratories
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _

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RE: [Histonet] Uneven Staining - Update

[Elizabeth Cameron]

This morning I decided to pH our DI water, which came out higher than it 
should.  I then did an experiment where I cut a kidney and floated it on three 
water baths - one a bit basic, one a bit acidic, and one neutral.  I cut two 
sections for each waterbath - one was picked up right away, and the other sat 
for 10 minutes (we occasionally need to let the kidneys sit to remove wrinkles, 
although not generally this long).  There was no difference between the 
sections that were picked up immediately and those that sat in the neutral 
waterbath.  The sections from the acidic waterbath did not stain well either 
way.  The sections from the basic water bath stained fine if they were picked 
up immediately, but did not stain well after sitting for 10 minutes, so I think 
we’ve figured out the problem.

Thanks for all the great suggestions!  It's so nice having access to so many 
knowledgeable histotechs when these strange issues arise!

-Liz

-Original Message-
From: Cristi Rigazio [mailto:cls71...@gmail.com] 
Sent: Tuesday, November 27, 2012 4:01 PM
To: Rene J Buesa
Cc: Elizabeth Cameron; histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] Uneven Staining

As most other bases have been covered, do you leave space between the slides in 
the rack?  Once in a blue moon we had stains from the same rack that differed 
in intensity, after much troubleshooting we started putting space between the 
slides and we haven't had the problem since.  Just an idea.
Cristi

Sent from my iPhone

On Nov 27, 2012, at 12:24 PM, Rene J Buesa  wrote:

> This is quite a puzzling situation.
> You have ruled out dewaxing (that was my first idea of cause) but I do not 
> think that soaking will be the problem because the sections are fixed and 
> infiltrated with paraffin, so there is no possibility of "diluting" in water.
> Thick/thin sections in a series is a very rare occurrence so I am also at a 
> loss with this problem. Weak staining solutions could cause it but all 
> sections in a series will be equally weak.
> Just in case, since you have covered the dewaxing, issue do the following: do 
> not leave the sections too long in the water bath and dry them in the oven as 
> soon as they are drained.
> René J.
> 
> From: Elizabeth Cameron 
> To: "histonet@lists.utsouthwestern.edu" 
> 
> Sent: Tuesday, November 27, 2012 1:35 PM
> Subject: [Histonet] Uneven Staining
> 
> About a year ago, I posted about issues with staining mouse kidney 
> sections (see below for original post).  We realized that part of the problem 
> was the sections drying during staining, which has been resolved, but we are 
> still having some issues with some slides staining well and others being very 
> pale.  This time it seems to be consistent across the slide, so I know it is 
> not a drying issue.  They are serial sections, so they are all cut together 
> and stained together, and they are from the same animal.  I don't believe 
> deparaffinization is an issue - we have tried longer times with fresh 
> solutions and it doesn't make a difference.  We have also tried re-staining 
> the slides after letting them sit in xylene overnight, and there is no 
> difference in the staining.  I am now thinking that it may be related to how 
> long the sections soak or how long they sit on the waterbath.  They are NBF 
> fixed, and there is a fine line between soaking well for hydration and 
> oversoaking, resulting in swelling.  Has anyone experienced anything like 
> this?  Any other ideas?  I am at a loss, and I'd really like for this to work!
> Thanks,
> Liz
> 
> We are doing a Hale's colloidal iron stain (no counterstain) on serial 
> sections of mouse kidneys.  We are staining an entire kidney at a time (about 
> 90-150 slides), and after many successful runs, we are now finding some 
> slides in each batch with very uneven staining.  Half of a section will stain 
> as it should, and the rest of the section is very pale.  It seems to be in a 
> similar area from one section to the next, but not exactly the same area.  It 
> does not look like a deparaffinization issue.  There may be two or three 
> slides in a row like this, or just one section on a slide, followed by 30-40 
> that look fine.  The sections are 6 microns. Any ideas on why this might be 
> happening?  Thanks!
> 
> 
> 
> The information in this email, including attachments, may be confidential and 
> 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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The information in this email, including attachments, may be confidential and 
is intended sole

[Histonet] Deadly Bladex system

[Davide Costanzo]

Does anyone out there use the Bladex system for applying blades to their
handles?

This unit was forced upon us at one place, required by the OSHA inspectors
and specifically told to use Bladex to improve safety in the workplace. I
have found that this system not only does not improve safety, but is
actually a very unsafe unit. Using Bladex, I have found, the risk of injury
is higher than any other means by which we have ever placed a blade on a
handle. And it is incredibly expensive to boot.

Any thoughts on this unit? Does anyone actually like it, or find it safe?


-- 
*David Costanzo, MHS, PA (ASCP)*
Project Manager
*Blufrog Path Lab Solutions*
9401 Wilshire Blvd. Ste 650
Beverly Hills, CA 90212
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[Histonet] Re: eosin in the processor

[Bob Richmond]

Supposedly eosin used to color small specimens, or put in the
processor, fluoresces enough in tissue sections to interfere with
fluorescent stains.

I've seen safranin O (from the microbiology lab) used to mark small
surgical specimens like GI biopsies. I don't know whether you can put
it in the processor or not. Supposedly safranin O isn't fluorescent,
though I haven't verified that.

Bob Richmond
Samurai Pathoogist
Maryville TN

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[Histonet] FW: Peloris Rapid Tissue Processor

[DeMint, Valerie S]

 

 



From: DeMint, Valerie S 
Sent: Tuesday, November 27, 2012 9:29 AM
To: 'histo...@lists.utsouthwester.edu'
Subject: FW: Peloris Rapid Tissue Processor

 

 

 



From: DeMint, Valerie S 
Sent: Monday, November 26, 2012 3:47 PM
To: 'histo...@lists.utsouthwester.edu'
Subject: Peloris Rapid Tissue Processor

Valerie S. DeMint, HT (ASCP)

Waukesha Memorial Hospital

Waukesha, WI 53188

 

I am wondering if anyone has experienced any intermittent, unexplained
processing problems with their Peloris rapid tissue processor.  I
thought our laboratory has had all the correct measures in place to
provide us with consistent, good tissue processing result but we
continue to have failed runs. Leica technical support cannot locate any
problems with our instrument.  Here's what we are "using and doing":
Careful attention is made so that all specimens are fully fixed prior to
starting a processing run. We use ethanol for dehydration. Bottles 3, 4
and 5 are dedicated to graded alcohols at the default concentrations of
70%, 80% and 90%.  We are a xylene lab. We are using the recommended
xylene protocols and following the tissue size recommendations for each
protocol.  We use the Activeflow cassettes that are designed to work
best with a rapid tissue processor.  We do not use anything like biopsy
pads that would affect carryover or flow of reagents.

Most recently, we had a "bad" run of medium size tissues that were over
processed.  The only reagent change prior to this run was the 70%
alcohol had been changed.  Following this bad processing run I checked
the alcohol concentration with a hydrometer and it read correctly.
There were no error codes on the instrument to indicate a problem.  

Any ideas or suggestions will be greatly appreciated.

 

 



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[Histonet] Quick Question...

[Pam Barker]

Hi Histonetters!!
I hope you are having a great day.  I have a quick question.  Is ASCP
certification required in order to work in the state of Michigan?

Happy Holidays !!

Thank You!
  
 
Pam Barker
President/Senior Recruiting Specialist-Histology
RELIA Solutions
Specialists in Allied Healthcare Recruiting
5703 Red Bug Lake Road #330
Winter Springs, FL 32708-4969
Phone: (407)657-2027
Cell: (407)353-5070
FAX: (407)678-2788
E-mail: rel...@earthlink.net 
www.facebook.com  /PamBarkerRELIA
www.linkedin.com/in/reliasolutions
www.twitter.com/pamatrelia 


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Re: [Histonet] Any reference labs offering CD52 (CAMPATH) on human FFPE specimens?

[Rob Day]

You might want to try www.phylogenyinc.com.


On Nov 28, 2012, at 10:54 AM, Sebree Linda A  wrote:

> Looking for a reference lab offering the above.
> 
> Thanks,
> 
> Linda A. Sebree
> 
> University of Wisconsin Hospital & Clinics
> IHC/ISH Laboratory
> 600 Highland Ave.
> Madison, WI 53792
> 
> (608)265-6596
> FAX: (608)262-7174
> 
> 
> ___
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[Histonet] Any reference labs offering CD52 (CAMPATH) on human FFPE specimens?

[Sebree Linda A]

Looking for a reference lab offering the above.

Thanks,

Linda A. Sebree

University of Wisconsin Hospital & Clinics
IHC/ISH Laboratory
600 Highland Ave.
Madison, WI 53792

(608)265-6596
FAX: (608)262-7174


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[Histonet] eosin on processor

[Steve McClain]

Eosin on processor can be done with dirty eosin waste saved from the stainer;
adding eosin powder sounds like a mess.

Especially helpful for orienting small samples with a sidedness, skin GI GU 
oral, cornea
No common down sides for routine HE, PAS, Trichrome, or IHC even with Red 
chromagens.
We leave it off of our research processors, just in case
Eosin conflicts with a few obscure
extraordinary stains with pale staining, e.g., Acridine Orange
or some fluorochromes (fluoroscein),
however, given variety of fluorochromes available, use another for your signal
and for those others eosin provides a roadmap to navigate the section.

Eosin can be made to stain several fungal structures.

Steve
Steve A. McClain, MD
McClain Labs, LLC 45 Manor Road, Smithtown, NY 11787 631 361 4000

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RE: [Histonet] eosin in the processor

[Davis, Cassie]

Hi Kim, 

try 10cc of the prediluted Eosin (you use in H & E staining) in your last 100% 
before your Xylene on the processor. By putting it in the last absolute it will 
not wash out. Your first Xylene will end up a little pink but your eyes will 
not strain as much trying to fine the tiny tissue fragments. (great for 
thread-like prostate biopsies too) We had been using it for years & our IHC 
seems fine. Make sure to keep your solutions for the clean cycles maintenanced 
on the automatic processors to avoid clogging. The only reason we stopped was 
the folks we purchased the new processor from told us using it would invalidate 
the warranty on the new processor.(I really miss it)

Cassandra Davis
cda...@che-east.org
302-575-8095


From: histonet-boun...@lists.utsouthwestern.edu 
[histonet-boun...@lists.utsouthwestern.edu] on behalf of Kim Merriam 
[kmerriam2...@yahoo.com]
Sent: Wednesday, November 28, 2012 7:54 AM
To: Histonet
Subject: [Histonet] eosin in the processor
Hi Everyone,
Years ago, my lab used to put eosin in the processor to lightly tint the 
smaller mouse tissues.  I can't remember which station we put it in (I think it 
was the 2nd 100% ethanol).  Also, back then my lab didn't do any IHC; will the 
eosin affect any IHC that might be done (I am guessing no, but I want to be 
sure).
Thanks,
Kim
Kim Merriam, MA, HT(ASCP)QIHC
Cambridge, MA
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Re: [Histonet] Waste drum labeling

[Rene J Buesa]

With different products the toxicity level will vary. Write in the diamond the 
highest toxic level even if that is not the most abundant.
By doing so you cover all inside the drum regarding their toxicity. I do not 
see the need for the emergency phone but if they told you to so so, do it.
René J.

From: Amber McKenzie 
To: 
Cc: "histonet@lists.utsouthwestern.edu"  
Sent: Wednesday, November 28, 2012 1:16 AM
Subject: [Histonet] Waste drum labeling

For EPA standards, how do you label your waste drums?  I have one 55 lb drum 
that we only pour ventana waste in.  What numbers do you write in the diamond 
to cover all the kits that you stain with?  2 reps from EPA came by my lab 
yesterday and said i needed an emergency phone list by our phone??  Does anyone 
have the checklist for EPA that small labs should follow?


From: histonet-boun...@lists.utsouthwestern.edu 
[histonet-boun...@lists.utsouthwestern.edu] on behalf of Cristi Rigazio 
[cls71...@gmail.com]
Sent: Tuesday, November 27, 2012 3:01 PM
To: Rene J Buesa
Cc: histonet@lists.utsouthwestern.edu; Elizabeth Cameron
Subject: Re: [Histonet] Uneven Staining

As most other bases have been covered, do you leave space between the slides in 
the rack?  Once in a blue moon we had stains from the same rack that differed 
in intensity, after much troubleshooting we started putting space between the 
slides and we haven't had the problem since.  Just an idea.
Cristi

Sent from my iPhone

On Nov 27, 2012, at 12:24 PM, Rene J Buesa  wrote:

> This is quite a puzzling situation.
> You have ruled out dewaxing (that was my first idea of cause) but I do not 
> think that soaking will be the problem because the sections are fixed and 
> infiltrated with paraffin, so there is no possibility of "diluting" in water.
> Thick/thin sections in a series is a very rare occurrence so I am also at a 
> loss with this problem. Weak staining solutions could cause it but all 
> sections in a series will be equally weak.
> Just in case, since you have covered the dewaxing, issue do the following: do 
> not leave the sections too long in the water bath and dry them in the oven as 
> soon as they are drained.
> René J.
>
> From: Elizabeth Cameron 
> To: "histonet@lists.utsouthwestern.edu" 
> Sent: Tuesday, November 27, 2012 1:35 PM
> Subject: [Histonet] Uneven Staining
>
> About a year ago, I posted about issues with staining mouse kidney sections 
> (see below for original post).  We realized that part of the problem was the 
> sections drying during staining, which has been resolved, but we are still 
> having some issues with some slides staining well and others being very 
> pale.  This time it seems to be consistent across the slide, so I know it is 
> not a drying issue.  They are serial sections, so they are all cut together 
> and stained together, and they are from the same animal.  I don't believe 
> deparaffinization is an issue - we have tried longer times with fresh 
> solutions and it doesn't make a difference.  We have also tried re-staining 
> the slides after letting them sit in xylene overnight, and there is no 
> difference in the staining.  I am now thinking that it may be related to how 
> long the sections soak or how long they sit on the waterbath.  They are NBF 
> fixed, and there is a fine line between soaking well for
> hydration and oversoaking, resulting in swelling.  Has anyone experienced 
> anything like this?  Any other ideas?  I am at a loss, and I'd really like 
> for this to work!
> Thanks,
> Liz
>
> We are doing a Hale's colloidal iron stain (no counterstain) on serial 
> sections of mouse kidneys.  We are staining an entire kidney at a time (about 
> 90-150 slides), and after many successful runs, we are now finding some 
> slides in each batch with very uneven staining.  Half of a section will stain 
> as it should, and the rest of the section is very pale.  It seems to be in a 
> similar area from one section to the next, but not exactly the same area.  It 
> does not look like a deparaffinization issue.  There may be two or three 
> slides in a row like this, or just one section on a slide, followed by 30-40 
> that look fine.  The sections are 6 microns. Any ideas on why this might be 
> happening?  Thanks!
>
>
>
> The information in this email, including attachments, may be confidential and 
> 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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[Histonet] Histologist need in Austin TX.doc

[Esparza, Sandra E.]

Histologist needed in Austin TX


We have a full time day  position opened for a HT or HTL (ASCP) registered 
histologist; requires 3 years experience and strong embedding and cutting 
skills. Experience in muscle enzymes, EM and IHC preferred.   If you are 
interested in learning new techniques and working in a team environment, in a  
# 1 Trauma Children's hospital with state of the art equipment; in beautiful 
progressive Austin Texas please call Sandra Esparza BS, HT, QIHC at 
512-324- x87062  or fill out an  application at seton.org  (#040492 Dell 
Children's).
 Please note; we are not able to work with recruiters at this time.


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RE: [Histonet] eosin in the processor

[Monfils, Paul]

Eosin won't interfere with binding of antibodies, but eosin is
fluorescent.  You might want to consider that.



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RE: [Histonet] eosin in the processor

[gayle callis]

The only immunostaining any residual eosin in tissue might affect is if you
do immunofluorescence.  Eosin does fluoresce.   Others can address any
effect on IHC.  

Gayle Callis
HTL/HT/MT(ASCP)
Bozeman  MT 

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Kim Merriam
Sent: Wednesday, November 28, 2012 5:55 AM
To: Histonet
Subject: [Histonet] eosin in the processor

Hi Everyone,

Years ago, my lab used to put eosin in the processor to lightly tint the
smaller mouse tissues.  I can't remember which station we put it in (I think
it was the 2nd 100% ethanol).  Also, back then my lab didn't do any IHC;
will the eosin affect any IHC that might be done (I am guessing no, but I
want to be sure).

Thanks,
Kim
 
Kim Merriam, MA, HT(ASCP)QIHC
Cambridge, MA
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[Histonet] MSI antibodies

[lynch kenia]

Need help please..does anyone have a colon block, or would be will ing to 
cut me a few slides of colon, that has stained positive (which is actually no 
staining) for the MSI antibodies; MLH1,PMS2, MSH6, MSH2 ?
 
Thanks!
 
Kenia Lynch, M.T./H.T.
Histology/Molecular Supervisor
Caldwell Memorial Hospital
321 Mulberry Street SW
Lenoir, NC  28645
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[Histonet] Protocol for preparing Cell Blocks

[Kelly Colpitts]

Hi Histo World,
We are currently looking at how we process cell blocks and were wondering if 
anyone had a procedure they would be willing to share with us.  Currently, we 
spin the fluids down in formalin and put the entire button in a biowrap and 
process that.  This leaves our cell blocks very bloody and not easy to embed.  
I have processed cytology fluids in past jobs where I used cytolyte but I do 
not have a copy of that protocol.  If you would be so kind as to forward me 
what works for you, that would be great!  My email address is 
kelly.colpi...@nationwidechildrens.org
 
Thanks,
Kelly 
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RE: [Histonet] eosin in the processor

[Lynette Pavelich]

Of our 3- 100% ETOH on our processor, we put about 75mL of LIQUID eosin in our 
"dirtiest" 100%. Our purchased eosin is made with 95% alcohol, and it works 
very well, not affecting the processing.

We used to use about 1/4 tsp of Eosin Y powder in our "dirtiest" 100%, but as 
it needs some water to dissolve, it caused trouble in the valves of the 
processor (oops). We tried using it in our 95%, but it got too washed out by 
the end of processing. 

No ill effects on our IHC.

Lynette



From: histonet-boun...@lists.utsouthwestern.edu 
[histonet-boun...@lists.utsouthwestern.edu] on behalf of Kim Merriam 
[kmerriam2...@yahoo.com]
Sent: Wednesday, November 28, 2012 7:54 AM
To: Histonet
Subject: [Histonet] eosin in the processor

Hi Everyone,

Years ago, my lab used to put eosin in the processor to lightly tint the 
smaller mouse tissues.  I can't remember which station we put it in (I think it 
was the 2nd 100% ethanol).  Also, back then my lab didn't do any IHC; will the 
eosin affect any IHC that might be done (I am guessing no, but I want to be 
sure).

Thanks,
Kim

Kim Merriam, MA, HT(ASCP)QIHC
Cambridge, MA
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[Histonet] Workflow Consulting Opportunity

[Matt Ward]

Good morning,





We have had a new position open with a global leader in histology. Please
contact me directly at m...@personifysearch.com to learn more.





Manager, Workflow Consulting –East



The Company:



Our client is a leading developer and producer of innovative high-tech
precision optics systems for the analysis of microstructures.  As one of
the market leaders in each of the fields of Cancer Diagnostics, Anatomical
Pathology, Imaging Systems, Specimen Preparation and Medical Equipment.

Comprising nine manufacturing facilities in seven countries, sales and
service companies in 20 countries and an international network of dealers,
the company is represented in over 100 countries.



The Opportunity:



The company currently has an opening for an Manager, Workflow Consulting in
Cancer Diagnostics. The person who fills this position can live anywhere in
the Eastern US.  All applicants must not be adverse to travel, as this is a
position that may require you to travel when necessary.



Base: Commensurate with Experience + Bonus



Primary Responsibilities:



The primary responsibility of this role will be to achieve Company sales
and profitability goals by offering a value-added service to end-user that
provides workflow consulting services and lean principles to optimize
customer laboratories performance.  Drive change in anatomic pathology
laboratories utilizing Lean principles, information management, and
hardware/software solutions.  The solution for such a change, efficiency
gains and waste elimination is the Company product offering.



Additional Responsibilities:



- Achieve monthly, quarterly, annual unit and revenue goals for the
Division.  Track KPI to measure revenue generated through Lean Consulting
Services



- Analyze new and existing customer laboratory organizations and workflow
practices and recommend short and long-term improvements



- Utilize Company Business System tools to credibly recommend changes to
lab practices to eliminate waste, reduce cost and improve quality and
turnaround times



- Analyze and report market trends and innovative competitive activities
for Lean Services



- Working in conjunction with local sales representatives, Area Sales
Managers and Directors of Sales plan and schedule face-face account calls
to current and potential end-users.  Train Sales force on basic lean
principles for them to help market lean services and be able to follow-up
with customers



- In conjunction with Sales and Marketing, identify and develop key
accounts in the territory.  In conjunction with Director of Corporate
Accounts, manage national accounts within territory requiring corporate
coordination to enable closure and compliance of contracts



- Prepare monthly territory status reports on lean activities to Management
Maintain and report monthly on Workflow opportunities and projects



- Manage operating expenses within assigned budget



- Maintain technical, product, applications and sales skill knowledge.

Maintain current knowledge of competition and market through study of
competitive marketing information, competitive literature, and field
surveillance or competition



- Prospect for all product opportunities.  Follow-up on all sales leads
with status review immediately upon receipt



- Participate in sales meetings and national trade shows as appropriate and
authorized



- Conduct Lunch and Learns, workshops, seminars or focus groups at local
technical society meetings as appropriate and authorized



- Promote the Company as the pathology market leader in quality and
innovation





Education and Experience Required:







- BA/BS in Life Sciences or equivalent required



- MBA preferred but not required



- 2-5 years Histology/Pathology laboratory experience in clinical ,
research or industrial setting desirable but not required



- Understanding of pathology marketplace with strong technical acumen



- 2-5 years knowledge of Company Business Systems, Six Sigma, or Lean
Principles required selling experience or consumables



- Outstanding problem solving skills. Can manage multiple layers of
personnel within customer site 1-3 years Histology laboratory experience in
clinical, research or industrial setting desirable but not required



- 1-3 years of product management or sales experience in a related
discipline preferred but not required







Regards,





Matt Ward

*Account Executive*

*Personify*

5020 Weston Parkway Suite 315

Cary NC 27513

(Tel) 800.875.6188 direct ext 103

(Fax) 919.460.0642

 www.personifysearch.com
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[Histonet] eosin in the processor

[Kim Merriam]

Hi Everyone,

Years ago, my lab used to put eosin in the processor to lightly tint the 
smaller mouse tissues.  I can't remember which station we put it in (I think it 
was the 2nd 100% ethanol).  Also, back then my lab didn't do any IHC; will the 
eosin affect any IHC that might be done (I am guessing no, but I want to be 
sure).

Thanks,
Kim
 
Kim Merriam, MA, HT(ASCP)QIHC
Cambridge, MA
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RE: [Histonet] Waste drum labeling

[Cynthia Pyse]

I would also be interested in the answers to this.
Cindy 

Cindy Pyse, CLT, HT (ASCP)
Laboratory Manager
X-Cell Laboratories
20 Northpointe Parkway Suite 100
Amherst, NY 14228
716-250-9235 etx. 232
e-mail cp...@x-celllab.com

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Amber
McKenzie
Sent: Wednesday, November 28, 2012 1:16 AM
Cc: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Waste drum labeling

For EPA standards, how do you label your waste drums?  I have one 55 lb drum
that we only pour ventana waste in.  What numbers do you write in the
diamond to cover all the kits that you stain with?  2 reps from EPA came by
my lab yesterday and said i needed an emergency phone list by our phone??
Does anyone have the checklist for EPA that small labs should follow?


From: histonet-boun...@lists.utsouthwestern.edu
[histonet-boun...@lists.utsouthwestern.edu] on behalf of Cristi Rigazio
[cls71...@gmail.com]
Sent: Tuesday, November 27, 2012 3:01 PM
To: Rene J Buesa
Cc: histonet@lists.utsouthwestern.edu; Elizabeth Cameron
Subject: Re: [Histonet] Uneven Staining

As most other bases have been covered, do you leave space between the slides
in the rack?  Once in a blue moon we had stains from the same rack that
differed in intensity, after much troubleshooting we started putting space
between the slides and we haven't had the problem since.  Just an idea.
Cristi

Sent from my iPhone

On Nov 27, 2012, at 12:24 PM, Rene J Buesa  wrote:

> This is quite a puzzling situation.
> You have ruled out dewaxing (that was my first idea of cause) but I do not
think that soaking will be the problem because the sections are fixed and
infiltrated with paraffin, so there is no possibility of "diluting" in
water.
> Thick/thin sections in a series is a very rare occurrence so I am also at
a loss with this problem. Weak staining solutions could cause it but all
sections in a series will be equally weak.
> Just in case, since you have covered the dewaxing, issue do the following:
do not leave the sections too long in the water bath and dry them in the
oven as soon as they are drained.
> René J.
>
> From: Elizabeth Cameron 
> To: "histonet@lists.utsouthwestern.edu" 
> 
> Sent: Tuesday, November 27, 2012 1:35 PM
> Subject: [Histonet] Uneven Staining
>
> About a year ago, I posted about issues with staining mouse kidney 
> sections (see below for original post).  We realized that part of the
problem was the sections drying during staining, which has been resolved,
but we are still having some issues with some slides staining well and
others being very pale.  This time it seems to be consistent across the
slide, so I know it is not a drying issue.  They are serial sections, so
they are all cut together and stained together, and they are from the same
animal.  I don't believe deparaffinization is an issue - we have tried
longer times with fresh solutions and it doesn't make a difference.  We have
also tried re-staining the slides after letting them sit in xylene
overnight, and there is no difference in the staining.  I am now thinking
that it may be related to how long the sections soak or how long they sit on
the waterbath.  They are NBF fixed, and there is a fine line between soaking
well for hydration and oversoaking, resulting in swelling.  Has anyone
experienced anything like this?  Any other ideas?  I am at a loss, and I'd
really like for this to work!
> Thanks,
> Liz
>
> We are doing a Hale's colloidal iron stain (no counterstain) on serial
sections of mouse kidneys.  We are staining an entire kidney at a time
(about 90-150 slides), and after many successful runs, we are now finding
some slides in each batch with very uneven staining.  Half of a section will
stain as it should, and the rest of the section is very pale.  It seems to
be in a similar area from one section to the next, but not exactly the same
area.  It does not look like a deparaffinization issue.  There may be two or
three slides in a row like this, or just one section on a slide, followed by
30-40 that look fine.  The sections are 6 microns. Any ideas on why this
might be happening?  Thanks!
>
>
>
> The information in this email, including attachments, may be confidential
and 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.
> ___
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
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[Histonet] RE: Uneven Staining

[Lynette Pavelich]

I have had this problem on a GI recut!! The first time through, the staining 
was perfect. The pathologist requested recuts, and I was cutting surgicals, so 
I put it in the back of the line up on my already melting ice bath to cut 
later. The recut was terrible. Nuclear staining was very weak! The block soaked 
up too much water. BUT!! The ribbon got progressively better on the slide!! 
Now, this lead to another discovery. In lovely processing.especially with a 
small 2mm GI biopsy, it shouldn't matter how long it sat on the wet ice, 
right??! Now I'm thinking it is a processing issue, NOT my wet ice, or how long 
I let it soak..right?!! In lovely processing land (i.e. research!!), if the 
tissue was processed well, and wax was infiltrated properly, it wouldn't soak 
up water. Don't you think THAT is the problem, histo friends??

At first, I thought.ohit's the dewaxing, and then I thought it was 
something not tight or too tight on the microtome, but you said everyone has 
the same problem. Are they all the same type of microtomes, if not, then I'm 
leaning towards processing issuessimilar to my GI issue.

Lynette


From: histonet-boun...@lists.utsouthwestern.edu 
[histonet-boun...@lists.utsouthwestern.edu] on behalf of Debbie Faichney 
[d.a.faich...@stir.ac.uk]
Sent: Wednesday, November 28, 2012 5:31 AM
To: Elizabeth Cameron; Rittman, Barry R; histonet@lists.utsouthwestern.edu
Subject: [Histonet] RE: Uneven Staining

Liz,

Could it be due to the presence of residual wash water or previous solution 
diluting the stain to give patchiness?  On the same thread, insufficient 
agitation of the slides in the solutions?  You do not say if staining is by 
hand or automated as this may be a contributor.
Whilst I have no experience of the staining method in use, I stain by hand and 
therefore able to ensure the previous solution is well drained prior to placing 
in the next.  The slides are also dipped in and out( once or twice) to ensure 
the previous solution is washed off as well as allowing the stain to be evenly 
distributed  amongst the slides.

I hope you are able to resolve this frustrating issue.

Kind regards

Debbie

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Elizabeth 
Cameron
Sent: 27 November 2012 19:13
To: Rittman, Barry R; histonet@lists.utsouthwestern.edu
Subject: [Histonet] RE: Uneven Staining

Barry,
Thanks for the suggestion.  We are generally only able to get about 10-15 
sections in a ribbon due to the nature of the tissue, and the pale staining 
last for 30+ sections at times.  We are also cutting on multiple microtomes 
(one microtome/kidney, but MANY kidneys!), so all microtomes would have to be 
affected.
No ideas on this one, so I appreciate any far-reaching thoughts!
Thanks,
Liz

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Rittman, Barry R
Sent: Tuesday, November 27, 2012 1:46 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] RE: Uneven Staining

Elizabeth
as the sections are evenly stained but vary in their staining from slide to 
slide, one suggestion is that section thickness varies. I know that this is 
reaching but if you section a ribbon and then place this on the water bath and 
then cut the next portion of ribbon then perhaps the microtome has a problem 
with the pause between cutting.
If this is the case try cutting a long ribbon and then mounting the separate 
slides.
Again, know this is reaching but difficult to see what it could be.
Barry


From: histonet-boun...@lists.utsouthwestern.edu 
[histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Elizabeth Cameron 
[elizabeth.came...@jax.org]
Sent: Tuesday, November 27, 2012 12:35 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Uneven Staining

About a year ago, I posted about issues with staining mouse kidney sections 
(see below for original post).  We realized that part of the problem was the 
sections drying during staining, which has been resolved, but we are still 
having some issues with some slides staining well and others being very pale.  
This time it seems to be consistent across the slide, so I know it is not a 
drying issue.  They are serial sections, so they are all cut together and 
stained together, and they are from the same animal.  I don't believe 
deparaffinization is an issue - we have tried longer times with fresh solutions 
and it doesn't make a difference.  We have also tried re-staining the slides 
after letting them sit in xylene overnight, and there is no difference in the 
staining.  I am now thinking that it may be related to how long the sections 
soak or how long they sit on the waterbath.  They are NBF fixed, and there is a 
fine line between soaking well for hydration and oversoaking, resulting in 
swelling.  Has anyon

[Histonet] RE: Uneven Staining

[Debbie Faichney]

Liz,

Could it be due to the presence of residual wash water or previous solution 
diluting the stain to give patchiness?  On the same thread, insufficient 
agitation of the slides in the solutions?  You do not say if staining is by 
hand or automated as this may be a contributor.
Whilst I have no experience of the staining method in use, I stain by hand and 
therefore able to ensure the previous solution is well drained prior to placing 
in the next.  The slides are also dipped in and out( once or twice) to ensure 
the previous solution is washed off as well as allowing the stain to be evenly 
distributed  amongst the slides.

I hope you are able to resolve this frustrating issue.

Kind regards

Debbie 

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Elizabeth 
Cameron
Sent: 27 November 2012 19:13
To: Rittman, Barry R; histonet@lists.utsouthwestern.edu
Subject: [Histonet] RE: Uneven Staining

Barry,
Thanks for the suggestion.  We are generally only able to get about 10-15 
sections in a ribbon due to the nature of the tissue, and the pale staining 
last for 30+ sections at times.  We are also cutting on multiple microtomes 
(one microtome/kidney, but MANY kidneys!), so all microtomes would have to be 
affected.
No ideas on this one, so I appreciate any far-reaching thoughts!
Thanks,
Liz

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Rittman, Barry R
Sent: Tuesday, November 27, 2012 1:46 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] RE: Uneven Staining

Elizabeth
as the sections are evenly stained but vary in their staining from slide to 
slide, one suggestion is that section thickness varies. I know that this is 
reaching but if you section a ribbon and then place this on the water bath and 
then cut the next portion of ribbon then perhaps the microtome has a problem 
with the pause between cutting.
If this is the case try cutting a long ribbon and then mounting the separate 
slides.
Again, know this is reaching but difficult to see what it could be.
Barry


From: histonet-boun...@lists.utsouthwestern.edu 
[histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Elizabeth Cameron 
[elizabeth.came...@jax.org]
Sent: Tuesday, November 27, 2012 12:35 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Uneven Staining

About a year ago, I posted about issues with staining mouse kidney sections 
(see below for original post).  We realized that part of the problem was the 
sections drying during staining, which has been resolved, but we are still 
having some issues with some slides staining well and others being very pale.  
This time it seems to be consistent across the slide, so I know it is not a 
drying issue.  They are serial sections, so they are all cut together and 
stained together, and they are from the same animal.  I don't believe 
deparaffinization is an issue - we have tried longer times with fresh solutions 
and it doesn't make a difference.  We have also tried re-staining the slides 
after letting them sit in xylene overnight, and there is no difference in the 
staining.  I am now thinking that it may be related to how long the sections 
soak or how long they sit on the waterbath.  They are NBF fixed, and there is a 
fine line between soaking well for hydration and oversoaking, resulting in 
swelling.  Has anyone experienced anything like this?  Any other ideas?  I am 
at a loss, and I'd really like for this to work!
Thanks,
Liz

We are doing a Hale's colloidal iron stain (no counterstain) on serial sections 
of mouse kidneys.  We are staining an entire kidney at a time (about 90-150 
slides), and after many successful runs, we are now finding some slides in each 
batch with very uneven staining.  Half of a section will stain as it should, 
and the rest of the section is very pale.  It seems to be in a similar area 
from one section to the next, but not exactly the same area.  It does not look 
like a deparaffinization issue.  There may be two or three slides in a row like 
this, or just one section on a slide, followed by 30-40 that look fine.  The 
sections are 6 microns. Any ideas on why this might be happening?  Thanks!
>



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