[Histonet] RE: H&E staining question

[Tony Henwood (SCHN)]

You could try 10 minutes treatment with periodic acid prior to H&E staining 
(Luna's technique I think)

Regards 
Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) 
Laboratory Manager & Senior Scientist 
Tel: 612 9845 3306 
Fax: 612 9845 3318 
the children's hospital at westmead
Cnr Hawkesbury Road and Hainsworth Street, Westmead
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA 


-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Connolly, Brett 
M
Sent: Tuesday, 2 July 2013 11:08 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] H&E staining question

Histonetters -

We had some rat tissues that were removed at necropsy and immediately fixed in 
formalin for 48 hrs.  Since the tissues could not be processed after the 48 hr 
fixation they were transferred to 70% ETOH for about a week, then processed and 
stained with H&E.


The H&E staining looks faded/washed out when compared to previous rat tissue 
that was fixed for 48 hr and then processed and stained without the extended 
time in 70% ETOH. The reason for limiting the fixation to 48 hr in the more 
recent study was that we were doing some IHC stains and wanted to keep the same 
fixation time as the previous study.

I have extra unstained slides left over - any suggestions on how to get some 
better, crisper H&E results?  Has anyone observed the washed out H&E appearance 
from tissues stored in 70% ETOH.

Thanks,
Brett

Brett M. Connolly, Ph.D.
Principal Scientist, Imaging Dept.
Merck & Co., Inc.
PO Box 4, WP-44K
West Point, PA 19486
brett_conno...@merck.com
T- 215-652-2501
F- 215-993-6803






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[Histonet] RE: Immunohistochemistry TAT

[Tony Henwood (SCHN)]

Yes, I agree with most.

But remember a major part of dewaxing sections is to heat the sections to 
remove as much wax as possible. 
This allows the xylene or hydrocarbon dewaxing to be as efficient as it can.
Remember to be wary of drying temperature, too high and some antigens (eg S100, 
5D3, CMV) will be adversely affected (see Henwood, A., (2005) "Effect of Slide 
Drying at 80°C on Immunohistochemistry" J Histotechnol 28(1):45-46).

We have noticed that staining for S100 was weaker on sections that had not been 
heated for as long as our routine time (30minutes 64oC).

One wonders whether heated detergent dewaxing might be more efficient at 
removing wax (see Henwood, A. F., Prasad, L., & Bourke, V. M. (2013). "The 
application of heated detergent dewaxing and rehydration to techniques for the 
demonstration of fungi: a comparison to routine xylene-alcohol dewaxing" J 
Histotechnol 36(2):45-50)

It is also possible that different waxes (especially in older, "cured" blocks) 
have different de-waxing requirements.

Something to consider!


Regards 
Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) 
Laboratory Manager & Senior Scientist 
Tel: 612 9845 3306 
Fax: 612 9845 3318 
the children's hospital at westmead
Cnr Hawkesbury Road and Hainsworth Street, Westmead
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA 

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Sebree Linda A
Sent: Tuesday, 2 July 2013 10:42 PM
To: 'Karlisch, Patricia'; 'histonet@lists.utsouthwestern.edu'
Subject: [Histonet] RE: Immunohistochemistry TAT

Pat,

Right off the bat I can see 2 things that could speed things up for you.  We 
have Ultras and only dry our slides for 10 minutes at 60 d C.  If tissues don't 
stay on well, try some "Stay On" in your water bath; we're experimenting with 
that now.  Also, we spend way too much time ourselves searching for blocks.   
See if there is a way to have them organized immediately after sectioning the 
H&Es so they are quicker to find...we are struggling with this.

My 2 cents; good luck.

Linda A. Sebree

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

(608)265-6596
FAX: (608)262-7174
-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Karlisch, 
Patricia
Sent: Monday, July 01, 2013 4:36 PM
To: 'histonet@lists.utsouthwestern.edu'
Subject: [Histonet] Immunohistochemistry TAT

Histonetters,
I am looking for some help in getting IHC slides out in an 8 hour period of 
time.  Here is some information.

* We have a busy Immunohistochemistry lab  and the volume of requests 
keep increasing.  We offer approximately 150 different  IHC stains ( including 
ISH and Dual ISH for Her 2).  Generally, there are request for  >150- 200  IHC 
tests twice a day, not including negative and positive (same slide) controls.

* The techs pull an AM log at 6am and another at 1pm.  The 1pm log will 
be handled while waiting for the IHC's to be completed from the 6am run  and 
these will always be run overnight.

* The 6am log requires that one tech start cutting each block for 
multiple IHC stains.  (A search for blocks and the necessary control slide 
occurs by the same person).   The slides are kept in a 60 degree oven for about 
60 minutes.  Most of these slides will be ready by 2-3:30pm.
   Here is my question.  If we allowed requests for IHC stains to wait until  
9AM  how could we still get slides out by 3:30pm-4pm? In other words we would 
not have access to an LIS log until 9am so that the pathologists could order 
from the morning biopsies.  What is the best way to do this  with 3 Ventana 
Ultra's  and one Benchmark XT. The blocks ( usually 40-50) need to be cut; 
sit in an oven and then get labeled and placed on the Ventana's.   All longer 
stains such as ISH would be run overnight.
   The following  tasks makes the 3:30- 4pm 
deadline difficult:

* Searching for the blocks

* Cutting the blocks (40-50) onto control slides + negative control

* One hour in the oven to dry  (Can we use 30minutes?)

* Attaching labels to 150 slides and setting up instruments

* One microtome

* 2 techs/ some of the time
 Can you share your workflow with me if you do a similar volume of slides 
within an 8 hours period.  How would you staff the two IHC techs to gain the 
most efficiency.  As you know the fuller the instrument the longer the staining 
process.

   Thank you,
  Pat Karlisch, Supervisor
pkarli...@hmc.psu.edu
Tel   717-531-6072
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[Histonet] markers in specific species

[pruegg]

Can anyone recommend a good antibody made in rat that works in mouse tissue
that is a nuclear marker and another one that is a membrane marker?

 

Can anyone recommend a good antibody made in goat that works in mouse and
rat tissue that is a nuclear marker and another one that is a membrane
marker?

 

 

Patsy Ruegg, HT(ASCP)QIHC
Ruegg IHC Consulting

40864 E. Arkansas Ave

Bennett, CO 80102

H 303-644-4538

C 720-281-5406

  prueg...@hotmail.com 

  rueggihcconsultin...@outlook.com

 

 

 

 


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you are NOT the intended recipient please contact the sender and dispose of
this e-mail as soon as possible.

 

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[Histonet] RE: Histonet Digest, Vol 116, Issue 1

[Jennifer Garrigan]

Re: H&E Staining

We do not have a high volume of rat tissues and thus stain by hand. Generally 
we have increased the time in hematoxylin and eosin slightly because the tissue 
is so dense, even if cut at the same thickness as mouse tissue. We do long term 
storage in 70% EtOH and have not had any problems with staining after that.

Hope this may help

Jennifer

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of 
histonet-requ...@lists.utsouthwestern.edu
Sent: Tuesday, July 02, 2013 1:06 PM
To: histonet@lists.utsouthwestern.edu
Subject: Histonet Digest, Vol 116, Issue 1

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Contents of Histonet digest..."


Today's Topics:

   1. renin IHC (Amy Lee)
   2. specimen tracking systems (Stacy McLaughlin)
   3. Immunohistochemistry TAT (Karlisch, Patricia)
   4. DAB Testing (Lori Harris)
   5. If going to holiday/vacation, or to unsubscribe (Lee & Peggy Wenk)
   6. AW: [Histonet] Immunohistochemistry TAT (Gudrun Lang)
   7. RE: Immunohistochemistry TAT (Sebree Linda A)
   8. H&E staining question (Connolly, Brett M)
   9. RE: H&E staining question - some clarification (Connolly, Brett M)
  10. Re: H&E staining question (Colleen Forster)
  11. RE: H&E staining question (Bea DeBrosse-Serra)
  12. IHC Elastin stain (Sebree Linda A)
  13. RE: IHC Elastin stain (Houston, Ronald)
  14. RE: DAB Testing (Cindy Pyse)


--

Message: 1
Date: Mon, 1 Jul 2013 11:05:50 -0700 (PDT)
From: Amy Lee 
Subject: [Histonet] renin IHC
To: histonet 
Message-ID:
<1372701950.60879.yahoomail...@web126006.mail.ne1.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1

Hello,
I am looking for a renin antibody for IHC on FFPE rodent tissue. I searched 
around and couldn't?decide which one I should try.?Could anybody recommend one?
?
Thanks in advance.
?
Amy

--

Message: 2
Date: Mon, 1 Jul 2013 18:38:30 +
From: Stacy McLaughlin 
Subject: [Histonet] specimen tracking systems
To: "histonet@lists.utsouthwestern.edu"

Message-ID:


Content-Type: text/plain; charset="us-ascii"

Hello Histoland,
Wondering how many of you use specimen tracking systems (bar-coded) and what 
type of system you use.
Pros-cons of each system?
Thanks,
Stacy


--

Message: 3
Date: Mon, 1 Jul 2013 21:35:32 +
From: "Karlisch, Patricia" 
Subject: [Histonet] Immunohistochemistry TAT
To: "'histonet@lists.utsouthwestern.edu'"

Message-ID:

Content-Type: text/plain; charset="us-ascii"

Histonetters,
I am looking for some help in getting IHC slides out in an 8 hour period of 
time.  Here is some information.

* We have a busy Immunohistochemistry lab  and the volume of requests 
keep increasing.  We offer approximately 150 different  IHC stains ( including 
ISH and Dual ISH for Her 2).  Generally, there are request for  >150- 200  IHC 
tests twice a day, not including negative and positive (same slide) controls.

* The techs pull an AM log at 6am and another at 1pm.  The 1pm log will 
be handled while waiting for the IHC's to be completed from the 6am run  and 
these will always be run overnight.

* The 6am log requires that one tech start cutting each block for 
multiple IHC stains.  (A search for blocks and the necessary control slide 
occurs by the same person).   The slides are kept in a 60 degree oven for about 
60 minutes.  Most of these slides will be ready by 2-3:30pm.
   Here is my question.  If we allowed requests for IHC stains to wait until  
9AM  how could we still get slides out by 3:30pm-4pm? In other words we would 
not have access to an LIS log until 9am so that the pathologists could order 
from the morning biopsies.  What is the best way to do this  with 3 Ventana 
Ultra's  and one Benchmark XT. The blocks ( usually 40-50) need to be cut; 
sit in an oven and then get labeled and placed on the Ventana's.   All longer 
stains such as ISH would be run overnight.
   The following  tasks makes the 3:30- 4pm 
deadline difficult:

* Searching for the blocks

* Cutting the blocks (40-50) onto control slides + negative control

* One hour in the oven to dry  (Can we use 30minutes?)

* Attaching labels to 150 slides and setting up instruments

* One microtome

* 2 techs/ some of the time
 Can you share your workflow

[Histonet] Re: H&E staining question

[Teri Johnson]

Hi Brett,

I agree with the others, the storage in 70% ETOH is likely not the cause of the 
washed out appearance of the staining.

*   Did the lab that did the H&E staining run a daily control? How did that 
look? Perhaps the tap water they are using went a little funny? Perhaps the 
depar xylene needs refreshing? The hematoxylin is used up? If all the other 
H&Es done that day look good, then look further.

*   Look at the fixative, it is possible to get bad batches of that. It is 
also possible to cram a lot of tissue into a small space resulting in 
inadequate fixation. Or perhaps it was bloody and not changed to fresh 
fixative? Was one sample fixed at room temperature and the other fixed at 4 
degrees C? Was the person doing the necropsy the same among animal batches?

*   Two things might help get better hematoxylin staining - I recall a 
journal article about using antigen retrieval pretreatment to improve H&E 
staining on samples that had been stored in fixative for a prolonged period of 
time. That might help.  Years ago,  I also noticed that the hematoxylin 
counterstained PAS slides looked better than our H&Es, so we put a bucket of 
0.5% periodic acid as an oxidation step before hematoxylin. It needs to be 
rinsed well so there is no periodic acid carryover (that'll kill your 
hematoxylin!), but that might improve your staining.

If you get this figured out, please let us know the cause and fix.

Teri Johnson
Manager, Histology
GNF - San Diego, CA
858-332-4752


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RE: [Histonet] DAB Testing

[Cindy Pyse]

I would be interested in this information as well.
Cindy

Cindy Pyse CLT, HT(ASCP)
Laboratory Manager
X-Cell Laboratories of WNY
20 Northpointe Parkway Ste 100
Amherst, NY 14228
716-250-9235 Ext. 232
cp...@x-celllab.com



-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Lori Harris
Sent: Monday, July 01, 2013 5:45 PM
To: 'histonet@lists.utsouthwestern.edu'
Subject: [Histonet] DAB Testing

Hello All!

Can someone recommend a company that does DAB waste testing? I would like to
get our waste tested from our Ventana Ultra. Thanks.

Lori A. Harris, HT (ASCP)
541-768-6078
Histology Section Lead
GSRMC Pathology Lab
3600 NW Samaritan Drive
Corvallis, OR 97330



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[Histonet] RE: IHC Elastin stain

[Houston, Ronald]

Linda,
we do it here


Ronnie Houston, MS HT(ASCP)QIHC
Anatomic Pathology Manager
ChildLab, a Division of Nationwide Children's Hospital
www.childlab.com

700 Children's Drive
Columbus, OH 43205
(P) 614-722-5450
(F) 614-722-2899
ronald.hous...@nationwidechildrens.org
www.NationwideChildrens.org

"One person with passion is better than forty people merely interested."
~ E.M. Forster



-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Sebree Linda A
Sent: Tuesday, July 02, 2013 12:00 PM
To: Histonet (Histonet@lists.utsouthwestern.edu)
Subject: [Histonet] IHC Elastin stain

Do any of you do this?  Do any referral labs offer this?

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] IHC Elastin stain

[Sebree Linda A]

Do any of you do this?  Do any referral labs offer this?

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] RE: H&E staining question

[Bea DeBrosse-Serra]

Hi Brett, 

That the H&E staining looks faded after storage in 70% seems very unlikely. We 
do this with rat and mouse tissues all the time. Did the other lab use a 
different Hematoxylin, Eosin, or used a different staining protocol on the 
autostainer?

Bea

Beatrice DeBrosse-Serra HT(ASCP)QIHC
Isis Pharmaceuticals
Antisense Drug Discovery
2855 Gazelle Ct.
Carlsbad, CA 92010
760-603-2371



-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Connolly, Brett 
M
Sent: Tuesday, July 02, 2013 6:08 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] H&E staining question

Histonetters -

We had some rat tissues that were removed at necropsy and immediately fixed in 
formalin for 48 hrs.  Since the tissues could not be processed after the 48 hr 
fixation they were transferred to 70% ETOH for about a week, then processed and 
stained with H&E.


The H&E staining looks faded/washed out when compared to previous rat tissue 
that was fixed for 48 hr and then processed and stained without the extended 
time in 70% ETOH. The reason for limiting the fixation to 48 hr in the more 
recent study was that we were doing some IHC stains and wanted to keep the same 
fixation time as the previous study.

I have extra unstained slides left over - any suggestions on how to get some 
better, crisper H&E results?  Has anyone observed the washed out H&E appearance 
from tissues stored in 70% ETOH.

Thanks,
Brett

Brett M. Connolly, Ph.D.
Principal Scientist, Imaging Dept.
Merck & Co., Inc.
PO Box 4, WP-44K
West Point, PA 19486
brett_conno...@merck.com
T- 215-652-2501
F- 215-993-6803






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http://www.merck.com/contact/contacts.html) that may be confidential, 
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Re: [Histonet] H&E staining question

[Colleen Forster]

I am interested in the answer as I use this same technique on my rodent 
samples and have not had this issue.





On 7/2/2013 8:07 AM, Connolly, Brett M wro

Histonetters -

We had some rat tissues that were removed at necropsy and immediately fixed in 
formalin for 48 hrs.  Since the tissues could not be processed after the 48 hr 
fixation they were transferred to 70% ETOH for about a week, then processed and 
stained with H&E.

The H&E staining looks faded/washed out when compared to previous rat tissue 
that was fixed for 48 hr and then processed and stained without the extended time 
in 70% ETOH. The reason for limiting the fixation to 48 hr in the more recent study 
was that we were doing some IHC stains and wanted to keep the same fixation time as 
the previous study.

I have extra unstained slides left over - any suggestions on how to get some better, 
crisper H&E results?  Has anyone observed the washed out H&E appearance from 
tissues stored in 70% ETOH.

Thanks,
Brett

Brett M. Connolly, Ph.D.
Principal Scientist, Imaging Dept.
Merck & Co., Inc.
PO Box 4, WP-44K
West Point, PA 19486
brett_conno...@merck.com
T- 215-652-2501
F- 215-993-6803






Notice:  This e-mail message, together with any attachments, contains
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proprietary copyrighted and/or legally privileged. It is intended solely
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[Histonet] RE: H&E staining question - some clarification

[Connolly, Brett M]

I forgot to mention that the H&E was performed in another lab at our 
facility...most likely on an autostainer.  We just did the IHC studies and I 
have always used 70% ETOH for tissue storage ... never heard of this staining 
phenomenon happening before.

Brett

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Connolly, Brett 
M
Sent: Tuesday, July 02, 2013 9:08 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] H&E staining question

Histonetters -

We had some rat tissues that were removed at necropsy and immediately fixed in 
formalin for 48 hrs.  Since the tissues could not be processed after the 48 hr 
fixation they were transferred to 70% ETOH for about a week, then processed and 
stained with H&E.


The H&E staining looks faded/washed out when compared to previous rat tissue 
that was fixed for 48 hr and then processed and stained without the extended 
time in 70% ETOH. The reason for limiting the fixation to 48 hr in the more 
recent study was that we were doing some IHC stains and wanted to keep the same 
fixation time as the previous study.

I have extra unstained slides left over - any suggestions on how to get some 
better, crisper H&E results?  Has anyone observed the washed out H&E appearance 
from tissues stored in 70% ETOH.

Thanks,
Brett

Brett M. Connolly, Ph.D.
Principal Scientist, Imaging Dept.
Merck & Co., Inc.
PO Box 4, WP-44K
West Point, PA 19486
brett_conno...@merck.com
T- 215-652-2501
F- 215-993-6803






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[Histonet] H&E staining question

[Connolly, Brett M]

Histonetters -

We had some rat tissues that were removed at necropsy and immediately fixed in 
formalin for 48 hrs.  Since the tissues could not be processed after the 48 hr 
fixation they were transferred to 70% ETOH for about a week, then processed and 
stained with H&E.

The H&E staining looks faded/washed out when compared to previous rat tissue 
that was fixed for 48 hr and then processed and stained without the extended 
time in 70% ETOH. The reason for limiting the fixation to 48 hr in the more 
recent study was that we were doing some IHC stains and wanted to keep the same 
fixation time as the previous study.

I have extra unstained slides left over - any suggestions on how to get some 
better, crisper H&E results?  Has anyone observed the washed out H&E appearance 
from tissues stored in 70% ETOH.

Thanks,
Brett

Brett M. Connolly, Ph.D.
Principal Scientist, Imaging Dept.
Merck & Co., Inc.
PO Box 4, WP-44K
West Point, PA 19486
brett_conno...@merck.com
T- 215-652-2501
F- 215-993-6803






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[Histonet] RE: Immunohistochemistry TAT

[Sebree Linda A]

Pat,

Right off the bat I can see 2 things that could speed things up for you.  We 
have Ultras and only dry our slides for 10 minutes at 60 d C.  If tissues don't 
stay on well, try some "Stay On" in your water bath; we're experimenting with 
that now.  Also, we spend way too much time ourselves searching for blocks.   
See if there is a way to have them organized immediately after sectioning the 
H&Es so they are quicker to find...we are struggling with this.

My 2 cents; good luck.

Linda A. Sebree

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

(608)265-6596
FAX: (608)262-7174
-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Karlisch, 
Patricia
Sent: Monday, July 01, 2013 4:36 PM
To: 'histonet@lists.utsouthwestern.edu'
Subject: [Histonet] Immunohistochemistry TAT

Histonetters,
I am looking for some help in getting IHC slides out in an 8 hour period of 
time.  Here is some information.

* We have a busy Immunohistochemistry lab  and the volume of requests 
keep increasing.  We offer approximately 150 different  IHC stains ( including 
ISH and Dual ISH for Her 2).  Generally, there are request for  >150- 200  IHC 
tests twice a day, not including negative and positive (same slide) controls.

* The techs pull an AM log at 6am and another at 1pm.  The 1pm log will 
be handled while waiting for the IHC's to be completed from the 6am run  and 
these will always be run overnight.

* The 6am log requires that one tech start cutting each block for 
multiple IHC stains.  (A search for blocks and the necessary control slide 
occurs by the same person).   The slides are kept in a 60 degree oven for about 
60 minutes.  Most of these slides will be ready by 2-3:30pm.
   Here is my question.  If we allowed requests for IHC stains to wait until  
9AM  how could we still get slides out by 3:30pm-4pm? In other words we would 
not have access to an LIS log until 9am so that the pathologists could order 
from the morning biopsies.  What is the best way to do this  with 3 Ventana 
Ultra's  and one Benchmark XT. The blocks ( usually 40-50) need to be cut; 
sit in an oven and then get labeled and placed on the Ventana's.   All longer 
stains such as ISH would be run overnight.
   The following  tasks makes the 3:30- 4pm 
deadline difficult:

* Searching for the blocks

* Cutting the blocks (40-50) onto control slides + negative control

* One hour in the oven to dry  (Can we use 30minutes?)

* Attaching labels to 150 slides and setting up instruments

* One microtome

* 2 techs/ some of the time
 Can you share your workflow with me if you do a similar volume of slides 
within an 8 hours period.  How would you staff the two IHC techs to gain the 
most efficiency.  As you know the fuller the instrument the longer the staining 
process.

   Thank you,
  Pat Karlisch, Supervisor
pkarli...@hmc.psu.edu
Tel   717-531-6072
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AW: [Histonet] Immunohistochemistry TAT

[Gudrun Lang]

Dear Pat,
here some inputs. If usefull, you have to decide for yourself.

Ad controls: We use for most of our antibodies an "universal-control". This
is put together with tiny pieces of normal tonsil, appendix, skin, liver and
pancreas. If you are interested, look at the website of NordiQC.
Some labs prefer to cut the controls in advance. This can also save time,
but the controls shouldn't get older than a week.
Ad speed: we cut the urgent cases in advance besides the HE with 6 slides
per block. These are needle biopsies from breast, liver, kidney,  etc.;
the pathologists only have to order a panel.
Ad drying: we often skip the drying in the oven at all. But it is important
to get rid of the water under the sections. (Besides: drying at high temp
for longer period can alter stainability.)
Ad LIS: we also have no LIS, but a light-version of it. We use a
WORD-document as form. The pathologists can mark the wished antibodies and
print the sheet directly onto our printer in the lab.
Ad organisation: isn't it possible with 4 instruments to keep the same
antibodies on each instrument? For example: one machine is the
breast-machine, one for lymphomas, etc. If this is possible, you can add a
"machine-code" on the labels and only look at this code while filling the
machines.

We work only with one Ultra. First technician comes at 6.30 and starts the
first run until 7.00-7.30. Second run is started at 10.30-11.00 and the run
over night is started around 15.00. The most slides have to been cut at
afternoon. And if there are more than 30, they go into the morning run.
This gives for the urgent biopsies an IHC-TAT about 5-6 hours (from HE
delivering to IHC delivering). Most other cases are ready over night (HE
delivering till 12.30 and IHC-ordering-deadline at 14.30).

Hope this helps
Gudrun Lang

Ltd. BMA
Histolabor Akh Linz, Austria

-Ursprüngliche Nachricht-
Von: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von Karlisch,
Patricia
Gesendet: Montag, 01. Juli 2013 23:36
An: 'histonet@lists.utsouthwestern.edu'
Betreff: [Histonet] Immunohistochemistry TAT

Histonetters,
I am looking for some help in getting IHC slides out in an 8 hour period
of time.  Here is some information.

* We have a busy Immunohistochemistry lab  and the volume of
requests keep increasing.  We offer approximately 150 different  IHC stains
( including ISH and Dual ISH for Her 2).  Generally, there are request for
>150- 200  IHC tests twice a day, not including negative and positive (same
slide) controls.

* The techs pull an AM log at 6am and another at 1pm.  The 1pm log
will be handled while waiting for the IHC's to be completed from the 6am run
and these will always be run overnight.

* The 6am log requires that one tech start cutting each block for
multiple IHC stains.  (A search for blocks and the necessary control slide
occurs by the same person).   The slides are kept in a 60 degree oven for
about 60 minutes.  Most of these slides will be ready by 2-3:30pm.
   Here is my question.  If we allowed requests for IHC stains to wait until
9AM  how could we still get slides out by 3:30pm-4pm? In other words we
would not have access to an LIS log until 9am so that the pathologists could
order from the morning biopsies.  What is the best way to do this  with 3
Ventana Ultra's  and one Benchmark XT. The blocks ( usually 40-50) need
to be cut; sit in an oven and then get labeled and placed on the Ventana's.
All longer stains such as ISH would be run overnight.
   The following  tasks makes the 3:30- 4pm
deadline difficult:

* Searching for the blocks

* Cutting the blocks (40-50) onto control slides + negative control

* One hour in the oven to dry  (Can we use 30minutes?)

* Attaching labels to 150 slides and setting up instruments

* One microtome

* 2 techs/ some of the time
 Can you share your workflow with me if you do a similar volume of
slides within an 8 hours period.  How would you staff the two IHC techs to
gain the most efficiency.  As you know the fuller the instrument the longer
the staining process.

   Thank you,
  Pat Karlisch, Supervisor
pkarli...@hmc.psu.edu
Tel   717-531-6072
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[Histonet] If going to holiday/vacation, or to unsubscribe

[Lee & Peggy Wenk]

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Thank you.

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