Re: [Histonet] Coverslipper

[Margaryan, Naira via Histonet]

Happy Thursday, 

We would like to sell our slightly used but in excellent condition Dako 
Coverslipper which was purchased in 2010 (purchase price was $ 25K). The 
instrument can handle up to 600 slides per hour making it one of the fastest on 
the market. In addition to the flexibility of the Coverslipper it is easy and 
straightforward to operate and cleaning and maintenance is simple to do. It is 
small enough to fit into fume cabinets, easy to move around and accepts a 
variety of commercial mounting media.

Picture by request.

Will take any offer,
Naira


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] X-gal staining

[Margaryan, Naira via Histonet]

Hello histonetters,

I have request from a scientist to perform X-gal staining on the formalin fixed 
paraffin-embedded kidney.
I was wondering if there is any possibility to do this staining on FFPE tissue 
as well as if there any possibility to perform a peroxidase DAB  staining for 
the X-gal? If so, may I ask you for a detailed protocol to perform this 
staining.
Any input and explanation why it is impossible is appreciated.


Thanks in advance,
Naira

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] job search in Chicago area

[Margaryan, Naira via Histonet]

Hello histonetters,

Please share this email with your colleagues.
I am in a job searching process in Chicago area to be close to my family as my 
appointment in West Virginia University Research Corporation, Morgantown, WV is 
coming to an end since my long-standing supervisor is closing her research 
laboratory.

I have had the privilege of establishing a Research Histology Core in Stanley 
Manne Children's Research Institute, Chicago, IL as well as a Research 
Histology Service in West Virginia University Research Corporation, Morgantown, 
WV, where I have provided histology services to scientists in various 
disciplines to support their research projects. I have also trained students 
enrolled in health science programs in the areas of routine histology and 
animal work.

My experience in cancer research with background in histology and in veterinary 
medicine has given me a unique opportunity to translate my knowledge from basic 
science to clinically relevant applications for diagnostic, prognostic and 
therapeutic development as they relate to cancer. My contributions to various 
research projects have included in vitro and in vivo work of predictive cancer 
biomarkers. I have been involved in testing novel humanized targeted 
antibody-based therapy in the treatment of human cancer in xenograft mouse 
models.

I am confident that my skills and experience would allow me to be a great fit 
for the research laboratory where I could provide excellent support for a wide 
range of scientific projects. I am a fast learner, a hard worker and a strong 
team player.
Furthermore, I am able to transfer a DAKO autostainer and an H station as 
part of my relocation.

My Resume and Curriculum Vitae are available by request and provide the more 
comprehensive list of my qualifications and accomplishments. I am very 
interested in meeting to discuss how I can best contribute to the work in 
research laboratory.

Sincerely,
Naira Margaryan

Naira V. Margaryan, D.V.M., Ph.D.
Senior Research Scientist
West Virginia University
Robert C. Byrd Health Sciences Center
Department of Biochemistry
PO Box 9142
200 Erma Byrd Biomedical Research Bldg.
108 Biomedical Road
Morgantown, WV 26506
Tel: 304-293-2213
naira.margar...@hsc.wvu.edu<mailto:naira.margar...@hsc.wvu.edu>

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Re: [Histonet] Tissue scrolling

[Margaryan, Naira via Histonet]

I do the same way! Good luck!
Naira


From: Colleen Forster 
Sent: Sunday, September 8, 2019 12:43 PM
To: John Garratt 
Cc: Erin McCarthy ; histonet-request 

Subject: Re: [Histonet] Tissue scrolling

We cut alot if these. This is the protocol:

1. The microtome, blade and water bath are cleaned well. We follow
withRNAse Away wipe down as an added precaution.

2. We use a coplin jar of 100% ethanol for forceps, pick or whatever other
tool you might  use that comes into contact with and dip and clean in
between each sample. I actually have 2 sets and put them into the alcohol
and switch out each time.

3 I use the RNAse free tubes to put the curls into.

4. You must wear gloves.

5. If they don't pick the samples up right away they are placed in a -20
freezer for storage.

They have good results.

Respectfully,

Colleen Forster HT(ASCP)
U of MN


On Sun, Sep 8, 2019, 10:36 AM John Garratt via Histonet <
histonet@lists.utsouthwestern.edu> wrote:

> The best I can offer is to advise you to wipe the knife holder area down
> with a cleaning solvent and use a fresh blade. Clean forceps with a solvent
> and place the scroll (50 microns) directly into a labelled 1.5mL plastic
> vial. I would be interested on other thoughts on this.
>
> John
>
>
>
> www.ciqc.ca
>
>
>
>
> ‐‐‐ Original Message ‐‐‐
> On Friday, September 6, 2019 3:05 PM, Erin McCarthy via Histonet <
> histonet@lists.utsouthwestern.edu> wrote:
>
> > Hello Histonetters,
> >
> > I am curious if any of you perform scrolling of paraffin embedded tissue
> > for molecular studies? If so, what precautions do you take to minimize
> > contamination? I am asking specifically about at the cutting station as
> > that is my area of expertise. But we are looking to pilot this for
> samples
> > that in the past have had large quantities of recuts.
> >
> > Thank you for any information you can provide!
> >
> >
> --
> >
> > Erin McCarthy, HT (ASCP)
> > Histology Supervisor
> >
> > Tempus Labs
> > 600 W. Chicago Ave.
> > Chicago IL 60654
> > Cell: (708)269-8610
> >
> >
> --
> >
> > This email and any attachments may contain privileged and confidential
> > information and/or protected health information (PHI) that is protected
> by
> > federal and state privacy laws.  It is intended solely for the use of
> > Tempus Labs and the recipient(s) named above.  Nothing contained in this
> > communication and any attachments thereto is intended to waive any
> > privileges or rights of confidentiality.  If you are not the recipient,
> or
> > the employee or agent responsible for delivering this message to the
> > intended recipient, you are hereby notified that any review,
> dissemination,
> > distribution, printing or copying of this email message and/or any
> > attachments is strictly prohibited. * If you have received this
> > transmission in error, please notify us immediately at (855)-442-8305
> > and permanently delete this email and any attachments*.
> >
> > Histonet mailing list
> > Histonet@lists.utsouthwestern.edu
> > http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
> ___
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Re: [Histonet] IHC mouse optomization

[Margaryan, Naira via Histonet]

Biocare Medical has pure ready to use secondary.. 

Naira 

-Original Message-
From: Blanca Lopez  
Sent: Wednesday, June 12, 2019 10:57 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] IHC mouse optomization

Morning,
I need a good advice in optimizing Mouse antibodies for research purposes. I 
use Dako/Agilent products. COX6B2 from Sigma is giving us hard times. Dako kit 
HRP has a mixture of mouse and rabbit so we think that might be the reason 
become negative.
Is there any tips for optimizing antibodies special for mouse tissue? Or if you 
can share your best procedures in IHC mouse stains. I can have a different ways 
and products to try. Any suggestion or opinions count. If you have any website 
that I can learn more about IHC for research mainly done in  Xenograft.
Thank you everybody:)

Blanca Lopez HT (ASCP)cm
Senior Histotechnologist
UT Southwestern Medical Center
Harold C. Simmons Comprehensive Cancer Center UTSTR Biorepository Tissue Lab
6000 Harry Hines Blvd NB5.102
Dallas, Texas 75390
214-648-7598
blanca.lo...@utsouthwestern.edu





UT Southwestern


Medical Center



The future of medicine, today.



___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Re: [Histonet] Coverslipper

[Margaryan, Naira via Histonet]

Happy Thursday, 

We would like to sell our slightly used but in excellent condition Dako 
Coverslipper which was purchased in 2010 just for $10K (purchase price was $ 
25K). The instrument can handle up to 600 slides per hour making it one of the 
fastest on the market. In addition to the flexibility of the Coverslipper it is 
easy and straightforward to operate and cleaning and maintenance is simple to 
do. It is small enough to fit into fume cabinets, easy to move around and 
accepts a variety of commercial mounting media.

Will take any offer,
Naira


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Re: [Histonet] Coverslipper

[Margaryan, Naira via Histonet]

Happy Friday, 

We would like to sell our slightly used but in excellent condition Dako 
Coverslipper which was purchased in 2010 $10K (purchase price was $ 25K). The 
instrument can handle up to 600 slides per hour making it one of the fastest on 
the market. In addition to the flexibility of the Coverslipper it is easy and 
straightforward to operate and cleaning and maintenance is simple to do. It is 
small enough to fit into fume cabinets, easy to move around and accepts a 
variety of commercial mounting media.

Will take any offer,
Naira


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Re: [Histonet] Coverslipper

[Margaryan, Naira via Histonet]

Happy Thursday, 

We would like to sell our slightly used but in excellent condition Dako 
Coverslipper which was purchased in 2010 $10K (purchase price was $ 25K). The 
instrument can handle up to 600 slides per hour making it one of the fastest on 
the market. In addition to the flexibility of the Coverslipper it is easy and 
straightforward to operate and cleaning and maintenance is simple to do. It is 
small enough to fit into fume cabinets, easy to move around and accepts a 
variety of commercial mounting media.

Will take any offer,
Naira


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] Polyclonal Rabbit Anti-Cytokeratin

[Margaryan, Naira via Histonet]

Hello,

I was asked to do IHC using the Polyclonal Rabbit Anti-Cytokeratin, Wide 
Spectrum Screening, Polyclonal, Unconjugated, Ig fraction, (Z062201-2).

Ca anyone help me with AR to use and concentration to start.

Thanks in advance,
Naira

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Re: [Histonet] Coverslipper

[Margaryan, Naira via Histonet]

Happy Thursday, 

We would like to sell our slightly used but in excellent condition Dako 
Coverslipper which was purchased in 2010 $10K (purchase price was $ 25K). The 
instrument can handle up to 600 slides per hour making it one of the fastest on 
the market. In addition to the flexibility of the Coverslipper it is easy and 
straightforward to operate and cleaning and maintenance is simple to do. It is 
small enough to fit into fume cabinets, easy to move around and accepts a 
variety of commercial mounting media.

Picture will send by request.

Will take any offer,
Naira


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] coverslipper CR10030

[Margaryan, Naira via Histonet]

Dear histinetters:

We can offer our slightly used DAKO coverslipper CR10030 for purchase.
Let us know if you are interested.

Thanks,
Naira


Ranked nationally in all 10 pediatric specialties by U.S. News & World Report  
(LCHOC Ver 1.0)


This message contains confidential information and is intended only for the 
individual named. If you are not the named addressee you should not 
disseminate, distribute or copy this e-mail. Please notify the sender 
immediately by e-mail if you have received this e-mail by mistake and delete 
this e-mail from your system. E-mail transmission cannot be guaranteed to be 
secure or error-free as information could be intercepted, corrupted, lost, 
destroyed, arrive late or incomplete, or contain viruses. The sender therefore 
does not accept liability for any errors or omissions in the contents of this 
message, which arise as a result of e-mail transmission. If verification is 
required please request a hard-copy version.  (LCHOC VER 1.0)

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] PD-L1 22c3 for IHC (paraffin)

[Margaryan, Naira via Histonet]

Dear Friends,

Can any of you share with me a full detailed protocol (time and dilutions) 
after deparaffinization for the PD-L1 22c3 for IHC (paraffin) to use on DAKO 
(not Link 48) autostainer?

http://www.agilent.com/en-us/products/pharmdx/pd-l1-ihc-22c3-pharmdx/pd-l1-ihc-22c3-pharmdx-for-autostainer-link-48-sk00621-5

Thanks in advance and have a nice weekend,
Naira


Ranked nationally in all 10 pediatric specialties by U.S. News & World Report  
(LCHOC Ver 1.0)


This message contains confidential information and is intended only for the 
individual named. If you are not the named addressee you should not 
disseminate, distribute or copy this e-mail. Please notify the sender 
immediately by e-mail if you have received this e-mail by mistake and delete 
this e-mail from your system. E-mail transmission cannot be guaranteed to be 
secure or error-free as information could be intercepted, corrupted, lost, 
destroyed, arrive late or incomplete, or contain viruses. The sender therefore 
does not accept liability for any errors or omissions in the contents of this 
message, which arise as a result of e-mail transmission. If verification is 
required please request a hard-copy version.  (LCHOC VER 1.0)

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] PD-L1 and TGFb for FFPE human tissue

[Margaryan, Naira via Histonet]

Good morning everyone,


I was wondering if anyone can give me an advise regarding the best Abs for the 
PD-L1 and TGFb for FFPE human tissue (except DAKO Abs, please).

Thanks in advance,
Naira



Ranked nationally in all 10 pediatric specialties by U.S. News & World Report  
(LCHOC Ver 1.0)


This message contains confidential information and is intended only for the 
individual named. If you are not the named addressee you should not 
disseminate, distribute or copy this e-mail. Please notify the sender 
immediately by e-mail if you have received this e-mail by mistake and delete 
this e-mail from your system. E-mail transmission cannot be guaranteed to be 
secure or error-free as information could be intercepted, corrupted, lost, 
destroyed, arrive late or incomplete, or contain viruses. The sender therefore 
does not accept liability for any errors or omissions in the contents of this 
message, which arise as a result of e-mail transmission. If verification is 
required please request a hard-copy version.  (LCHOC VER 1.0)

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] Immunoperoxidase Protocol on Cytospin

[Margaryan, Naira via Histonet]

Dear histonetters:

I never done ICC.

A scientist brought me a slide with Cytospined cells on for IHC.

I am going to fix with 10%NBF (it is only what I have now) then wash with TBST.

May I skip AR step?
May I use my usual IHC protocol and reagents which I usually use for FFPE 
tissue: blocks with H2O2, avidin/biotin/PB with TBST wash buffer

Thanks in advance,
Naira


Ranked nationally in all 10 pediatric specialties by U.S. News & World Report  
(LCHOC Ver 1.0)


This message contains confidential information and is intended only for the 
individual named. If you are not the named addressee you should not 
disseminate, distribute or copy this e-mail. Please notify the sender 
immediately by e-mail if you have received this e-mail by mistake and delete 
this e-mail from your system. E-mail transmission cannot be guaranteed to be 
secure or error-free as information could be intercepted, corrupted, lost, 
destroyed, arrive late or incomplete, or contain viruses. The sender therefore 
does not accept liability for any errors or omissions in the contents of this 
message, which arise as a result of e-mail transmission. If verification is 
required please request a hard-copy version.  (LCHOC VER 1.0)

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Re: [Histonet] Recycled reagents in VIP processor

[Margaryan, Naira via Histonet]

Absolutely agreed! 

Naira 

-Original Message-
From: Frazier, John [mailto:john.fraz...@roche.com] 
Sent: Tuesday, February 28, 2017 2:31 PM
To: Jay Lundgren <jaylundg...@gmail.com>
Cc: Margaryan, Naira <nmargar...@luriechildrens.org>; Gareth Davis 
<garethdavisy...@gmail.com>; histonet@lists.utsouthwestern.edu; Margaryan, 
Naira <naira.margar...@hsc.wvu.edu>
Subject: Re: [Histonet] Recycled reagents in VIP processor

As a six sigma consultant to histology laboratories, it has been my experience 
that recycling xylene and alcohol overall is not a cost saver. When you factor 
in both capital dollars and operational dollars, the savings is neutral. In 
addition to the neutral cost in recycling, you have to concern yourself with 
the quality of your in product on a daily basis. The last piece in the equation 
is the handling of Xylene with multiple touch points in between.  A movement 
within the histology world has begun with handling xylene (hazard
waste) as little as possible and/or reducing or eliminating its use where 
possible.

Sent from my iPhone

> On Feb 27, 2017, at 6:25 PM, Jay Lundgren <jaylundg...@gmail.com> wrote:
>
> When people say they are saving money by recycling reagents I always 
> wonder if they are including tech time (to run the still) in their 
> calculations.
>
> Sincerely,
>
> Jay A. Lundgren, M.S., HTL (ASCP)
>
> On Wed, Feb 22, 2017 at 2:32 PM, Margaryan, Naira via Histonet < 
> histonet@lists.utsouthwestern.edu> wrote:
>
>> WOULD LIKE TO KNOW AS WELL - IT IS WASTE OF MONEY
>>
>> Thanks,
>> Naira
>>
>>
>> Ranked nationally in all 10 pediatric specialties by U.S. News & 
>> World Report  (LCHOC Ver 1.0)
>>
>>
>> This message contains confidential information and is intended only 
>> for the individual named. If you are not the named addressee you 
>> should not disseminate, distribute or copy this e-mail. Please notify 
>> the sender immediately by e-mail if you have received this e-mail by 
>> mistake and delete this e-mail from your system. E-mail transmission 
>> cannot be guaranteed to be secure or error-free as information could 
>> be intercepted, corrupted, lost, destroyed, arrive late or incomplete, or 
>> contain viruses.
>> The sender therefore does not accept liability for any errors or 
>> omissions in the contents of this message, which arise as a result of 
>> e-mail transmission. If verification is required please request a 
>> hard-copy version.  (LCHOC VER 1.0)
>>
>> 
>> From: Gareth Davis via Histonet [histonet@lists.utsouthwestern.edu]
>> Sent: Wednesday, February 22, 2017 2:23 PM
>> To: histonet@lists.utsouthwestern.edu
>> Subject: [Histonet] Recycled reagents in VIP processor
>>
>> Hi,
>> I was always told not to use recycled reagents, i.e. Alcohol and 
>> Xylene, in processors.  I am using a VIP 300, refurbished, and I 
>> would rather not use recycled reagents in it.  But, during the last 
>> CAP inspection they suggested I use the recycled to save money.  And 
>> now my administration wants to cut cost.  Just wanted to know what labs were 
>> doing.
>> Thanks,
>>
>>
>> --
>> Ms. Gareth B. Davis, B.S., HT, QIHC (ASCP)cm Yuma Gastroenterology 
>> Yuma, AZ 85364
>> 928-248-5259
>> ___
>> Histonet mailing list
>> Histonet@lists.utsouthwestern.edu
>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>> ___
>> Histonet mailing list
>> Histonet@lists.utsouthwestern.edu
>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>>
>
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Re: [Histonet] Recycled reagents in VIP processor

[Margaryan, Naira via Histonet]

 WOULD LIKE TO KNOW AS WELL - IT IS WASTE OF MONEY

Thanks,
Naira


Ranked nationally in all 10 pediatric specialties by U.S. News & World Report  
(LCHOC Ver 1.0)


This message contains confidential information and is intended only for the 
individual named. If you are not the named addressee you should not 
disseminate, distribute or copy this e-mail. Please notify the sender 
immediately by e-mail if you have received this e-mail by mistake and delete 
this e-mail from your system. E-mail transmission cannot be guaranteed to be 
secure or error-free as information could be intercepted, corrupted, lost, 
destroyed, arrive late or incomplete, or contain viruses. The sender therefore 
does not accept liability for any errors or omissions in the contents of this 
message, which arise as a result of e-mail transmission. If verification is 
required please request a hard-copy version.  (LCHOC VER 1.0)


From: Gareth Davis via Histonet [histonet@lists.utsouthwestern.edu]
Sent: Wednesday, February 22, 2017 2:23 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Recycled reagents in VIP processor

Hi,
I was always told not to use recycled reagents, i.e. Alcohol and Xylene, in
processors.  I am using a VIP 300, refurbished, and I would rather not use
recycled reagents in it.  But, during the last CAP inspection they
suggested I use the recycled to save money.  And now my administration
wants to cut cost.  Just wanted to know what labs were doing.
Thanks,


--
Ms. Gareth B. Davis, B.S., HT, QIHC (ASCP)cm
Yuma Gastroenterology
Yuma, AZ 85364
928-248-5259
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] rat vomeronasal organ (VNO) fix, decal, proc. and paraffin embedding protocol

[Margaryan, Naira via Histonet]

Dear Histonet Colleagues,

I hope you had very nice and warm holiday season .

We are interested in a routinely used protocol to decal (if needed), process 
and paraffin embed the rat vomeronasal organ (VNO).

It would be greatly appreciated if anyone would be willing to share a protocol 
for post-harvest sample storage (i.e. PBS, 70% ETOH, saline, etc.), fixation, 
decalcification, processing before embedding.


Thanks in advance for your help!!

Naira


Ranked nationally in all 10 pediatric specialties by U.S. News & World Report  
(LCHOC Ver 1.0)


This message contains confidential information and is intended only for the 
individual named. If you are not the named addressee you should not 
disseminate, distribute or copy this e-mail. Please notify the sender 
immediately by e-mail if you have received this e-mail by mistake and delete 
this e-mail from your system. E-mail transmission cannot be guaranteed to be 
secure or error-free as information could be intercepted, corrupted, lost, 
destroyed, arrive late or incomplete, or contain viruses. The sender therefore 
does not accept liability for any errors or omissions in the contents of this 
message, which arise as a result of e-mail transmission. If verification is 
required please request a hard-copy version.  (LCHOC VER 1.0)

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Re: [Histonet] Ki67 antibody

[Margaryan, Naira]

Me too, love it!

Naira


Ranked nationally in all 10 pediatric specialties by U.S. News  World Report  
(LCHOC Ver 1.0)


This message contains confidential information and is intended only for the 
individual named. If you are not the named addressee you should not 
disseminate, distribute or copy this e-mail. Please notify the sender 
immediately by e-mail if you have received this e-mail by mistake and delete 
this e-mail from your system. E-mail transmission cannot be guaranteed to be 
secure or error-free as information could be intercepted, corrupted, lost, 
destroyed, arrive late or incomplete, or contain viruses. The sender therefore 
does not accept liability for any errors or omissions in the contents of this 
message, which arise as a result of e-mail transmission. If verification is 
required please request a hard-copy version.  (LCHOC VER 1.0)

-Original Message-
From: Tamara Howard [mailto:thow...@unm.edu]
Sent: Tuesday, June 09, 2015 12:30 PM
To: e...@georgetown.edu
Cc: histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] Ki67 antibody

Eva -


We use the SP6 rabbit monoclonal, formerly from NeoMarkers - they now belong to 
Thermo. Several companies sell this clone; I know AbCam also has it. It does 
require heat-mediated AR for FFPE tissues, but works beautifully in mouse  
human; I don't think I've used it on rat.


https://www.fishersci.com/shop/products/anti-ki-67-clone-sp6-thermo-scientific-lab-vision/p-3139002




Tamara




...
Tamara Howard
Dept. of Cell Biology  Physiology
University of New Mexico
Albuquerque, NM

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

RE: [Histonet] Re: squamous cell contamination on slides

[Margaryan, Naira]

Hi Hestonetters,

I need to do IHC of SC4MOL on FFPE sections.
If any of you have been done this staining before, please send me your 
procedure.

Thanks in advance,
Naira



Ranked nationally in all 10 pediatric specialties by U.S. News  World Report  
(LCHOC Ver 1.0)


This message contains confidential information and is intended only for the 
individual named. If you are not the named addressee you should not 
disseminate, distribute or copy this e-mail. Please notify the sender 
immediately by e-mail if you have received this e-mail by mistake and delete 
this e-mail from your system. E-mail transmission cannot be guaranteed to be 
secure or error-free as information could be intercepted, corrupted, lost, 
destroyed, arrive late or incomplete, or contain viruses. The sender therefore 
does not accept liability for any errors or omissions in the contents of this 
message, which arise as a result of e-mail transmission. If verification is 
required please request a hard-copy version.  (LCHOC VER 1.0)

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of gayle callis
Sent: Wednesday, October 29, 2014 1:48 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Re: squamous cell contamination on slides

After following this thread on a topic that has always been a pet peeve and a 
problem at times, a few suggestions. This is a problem discussed many times on 
Histonet.



If people are using the water bath as a finger bowl, they need to learn to 
not let fingers go snorkeling when picking up sections.  This is sloppy, poor 
technique.  Anyone can learn to never touch the water surface with bare skin. I 
have seen people immerse their fingers up to the first joint and their stained 
sections were covered by a snowstorm of squamous cells.



Be aware that squamous cells are going to exfoliate from other than hands, so 
not touching face and hair is good advice even if one wears gloves.



Hand lotion is helpful except for those slather on lotion or the lotion is 
particularly heavy duty and then they still touch the water. This can cause a 
double problem - an oil slick which is a terrible section adhesive along with 
squamous cells from bare skin.



Hold slide at top or on sides, as mentioned previously.

Wear gloves. Not always popular with a common argument one loses dexterity 
handling slides.  If gloves do not flop around loosely but fit the hand well, 
then dexterity is not lost.



Good luck



Gayle Callis

HTL/HT/MT(ASCP)




***



We have that problem mostly during the winter months when our hands get dry.
Use hand lotion, that usually helps a bit.



Thanks,



Michele Margiotta-Watz

Histology Supervisor

BMHMC

101 Hospital Rd.

Patchogue, NY 11772

631-654-7192



-Original Message-

From:  http://lists.utsouthwestern.edu/mailman/listinfo/histonet
histonet-bounces @t lists.utsouthwestern.edu [mailto:
http://lists.utsouthwestern.edu/mailman/listinfo/histonet histonet-bounces 
@t lists.utsouthwestern.edu] On Behalf Of Amber McKenzie

Sent: Tuesday, October 28, 2014 3:55 PM

To:  http://lists.utsouthwestern.edu/mailman/listinfo/histonet histonet @t 
lists.utsouthwestern.edu

Subject: [Histonet] Squamous cells staining on HE and IHC





Does anyone else have problems with what looks like squamous cells staining on 
your HE's and IHC's?  I'm trying to figure out how to eliminate that problem 
in our lab...wear gloves while cutting?  Change out water bath several times 
during shifts? Any suggestions?  Thanks!



___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

RE: [Histonet] Histology as art!

[Margaryan, Naira]

Hi Emily,

I don't mind if it is healthy tissue! It shows that even small cell or huge 
artery =all from the mother nature, don’t you think?

Naira 



Ranked nationally in all 10 pediatric specialties by U.S. News  World Report  
(LCHOC Ver 1.0)

This message contains confidential information and is intended only for the 
individual named. If you are not the named addressee you should not 
disseminate, distribute or copy this e-mail. Please notify the sender 
immediately by e-mail if you have received this e-mail by mistake and delete 
this e-mail from your system. E-mail transmission cannot be guaranteed to be 
secure or error-free as information could be intercepted, corrupted, lost, 
destroyed, arrive late or incomplete, or contain viruses. The sender therefore 
does not accept liability for any errors or omissions in the contents of this 
message, which arise as a result of e-mail transmission. If verification is 
required please request a hard-copy version.  (LCHOC VER 1.0)
-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Emily Brown
Sent: Friday, June 27, 2014 10:58 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Histology as art!

Hello Histonetters,

I'm really looking forward to going to the brand new Morbid Anatomy Museum in 
NYC, but imagine my surprise when I found some histology in their online gift 
shop!!
http://morbidanatomy.bigcartel.com/category/gifts
Histology is beautiful, but it is odd to look at those images on clothing.

Emily



By bitching and bitching and bitching, they could exhaust the drama of their 
own horror stories. Grow bored. Only then could they accept a new story for 
their lives. Move forward.

-Chuck Palahniuk, Haunted
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] FW: Hemoglobin

[Margaryan, Naira]

Hi histonetters,

Do you know how we can stain for hemoglobin in FFPE zebrafish tissue?

Thanks in advance,
Simone.


Ranked nationally in all 10 pediatric specialties by U.S. News  World Report  
(LCHOC Ver 1.0)

This message contains confidential information and is intended only for the 
individual named. If you are not the named addressee you should not 
disseminate, distribute or copy this e-mail. Please notify the sender 
immediately by e-mail if you have received this e-mail by mistake and delete 
this e-mail from your system. E-mail transmission cannot be guaranteed to be 
secure or error-free as information could be intercepted, corrupted, lost, 
destroyed, arrive late or incomplete, or contain viruses. The sender therefore 
does not accept liability for any errors or omissions in the contents of this 
message, which arise as a result of e-mail transmission. If verification is 
required please request a hard-copy version.  (LCHOC VER 1.0)

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

RE: [Histonet] Mouse lung IHC false positive/background issue

[Margaryan, Naira]

I have same problem for other Abs using in on mouse lung tissues... My 
secondary and tertiary have no problems on other tissues for negative controls 
as well

Naira
**
Confidentiality Note: This message (including any attachments) is intended for 
the use of the named recipient(s) only and may contain confidential and/or 
proprietary information. If you are not the intended recipient, please contact 
the sender and delete this message. Any unauthorized use of the information 
contained in this message is prohibited.


Ranked nationally in all 10 pediatric specialties by U.S. News  World Report  
(LCHOC Ver 1.0)

This message contains confidential information and is intended only for the 
individual named. If you are not the named addressee you should not 
disseminate, distribute or copy this e-mail. Please notify the sender 
immediately by e-mail if you have received this e-mail by mistake and delete 
this e-mail from your system. E-mail transmission cannot be guaranteed to be 
secure or error-free as information could be intercepted, corrupted, lost, 
destroyed, arrive late or incomplete, or contain viruses. The sender therefore 
does not accept liability for any errors or omissions in the contents of this 
message, which arise as a result of e-mail transmission. If verification is 
required please request a hard-copy version.  (LCHOC VER 1.0)
-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Lu Ze
Sent: Friday, October 11, 2013 2:38 PM
To: histonet@lists.utsouthwestern.edu
Cc: rebel...@gmail.com
Subject: [Histonet] Mouse lung IHC false positive/background issue

Hello everyone,

I forwarded message from my colleague, Rebecca (see below) to see if any one 
with IHC experience on mouse lung tissue have suggestion.


===

I'm having trouble with my IHC. I am staining for human MTDH (specific for 
human) on mouse tumor and lung sections from my xenograft mouse model. All of 
my tumors stained positive, but not the lung sections (which could possibly be 
the results). My real problem is that my negative control normal lung section 
stained positive for MTDH. I saw positive staining on the tissue near the 
outside and in the bronchioles.

I know the problem is not the secondary antibody because I did 2 slides (normal 
lung and previously positive tumor) without primary ab and they showed no 
positive staining.

My antibody is a rabbit monoclonal antibody at 1:200 for 1 hour. I'm blocking 
with 1% BSA in TBST for 1 hour. I'm using DAB substrate kit for developing.

What would you recommend?

Thank you,

Ze


Ze Lu, Ph.D.
Optimum Therapeutics, LLC
9363 Towne Centre Dr., Suite 110
San Diego, CA 92121
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] IHC (DAB) after HE?

[Margaryan, Naira]

Dear Histonetters,

I would like to perform IHC (DAB) after HE. Do I need to remove HE? If answer 
is YES, my question HOW? Can I ask the protocol for that?

Thanks in advance,
Naira

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] zebrafish and IHC

[Margaryan, Naira]

Dear Histonetters,

I am trying to do  IHC on FFPE 4 weeks old zebrafish with different Abs. Is 
there a trick working with zebrafish? I am using the same IHC protocol I always 
use on human and mouse tissue and my Abs are  suppose to work on fish as well 
as human. I run human and fish sections together with same AR and IHC protocol. 
By the end I get beautiful staining on human section and or nothing or some 
fuzzy/fuggy/unclarified/undistinguished reaction.

Any help is appreciated as I am new in fish field,
Naira

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] HE reagents

[Margaryan, Naira]

What companies HE reagents (catalog preferably) are the best for simple HE 
staining?



Thanks in advance,

Naira

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] proliferation and apoptosis

[Margaryan, Naira]

Hi histonetters,

I am looking for the good markers to detect (separately) proliferation and 
apoptosis of cells in tumor sections. Unfortunately, KI-67 stains apoptotic 
bodies as well as proliferated cells; and the tunnel assay shows both apoptotic 
body and proliferation.

Any suggestions for the FFPE tissue and the HRP protocol are appreciated.

Thanks in advance,
Naira

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] reagents without expiration dates

[Margaryan, Naira]

My oldest was from 1965, I can't bit Rene:)

Naira
 --

Message: 12
Date: Thu, 2 Feb 2012 13:14:53 -0800 (PST)
From: Rene J Buesa rjbu...@yahoo.com
Subject: Re: [Histonet] reagents without expiration dates

Ha.Ha,Ha!!!
I used to prepare staining solutions with some Merck-Darmstad anilines 
manufactured just after the Great War, i.e. the FIRST World War (about 1925 
before the World Great Depression).
Try to beat that!.
Ren?? J.

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] emergency question-IHC interruption

[Margaryan, Naira]

Hi,

Where I can stop my IHC, in what step?

I started deparaffinization, but I need to stop and run my IHC tomorrow. Can I 
leave my slides in 95% Ethanol, or Water or I have to do HIER? Where to store 
slides: in refrigerator or room.

Naira

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

FW: [Histonet] emergency question-IHC interruption

[Margaryan, Naira]

Thanks to all who are kindly and quick answered on my question.

Naira
On Thu, Oct 13, 2011 at 4:20 PM, Margaryan, Naira 
nmargar...@childrensmemorial.orgmailto:nmargar...@childrensmemorial.org 
wrote:
Hi,

Where I can stop my IHC, in what step?

I started deparaffinization, but I need to stop and run my IHC tomorrow. Can I 
leave my slides in 95% Ethanol, or Water or I have to do HIER? Where to store 
slides: in refrigerator or room.

Naira

___
Histonet mailing list
Histonet@lists.utsouthwestern.edumailto:Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] HE on cells, protocol

[Margaryan, Naira]

Hi Histonetters,

I have to do HE staining on cells that were growing up on coverslips. Can 
anyone send me a good protocol to fix these cells on same coverslip and HE 
staining, please?

Thanks in advance,
Naira

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] melanin counterstaining

[Margaryan, Naira]

Good morning,

I have heavily pigmented melanocytic cells and trying to differentiate between 
melanin pigment and chromogen (DAB) by using Azure B.

After IHC with DAB I incubate slides for 30-45 min in the working solution of:

 *   4ml of a 25mg/ml aqueous solution of Azure B
 *   3.4ml of 0.1M acetic acid
 *   600µl of 0.1M sodium acetate
 *   27ml of DI water
 *   5ml acetone

Then water wash and hematoxylin counter-stain. By the end all melanin supposed 
to turn blue-green, but I still see some remained brown melanin.

My question is: I would like to increase green color, so the concentration of 
what reagent I have to increase?

Thanks in advance,
Naira

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] Retirement

[Margaryan, Naira]

I am 15-18 years out from retirement also. Count me in as well

Naira



___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] How do you call...

[Margaryan, Naira]

I totally agree with Rene J.: there is anatomy and physiology becomes 
pathological that is why need to call Pathological Anatomy and Pathological 
Physiology

Foreign people learn what they hear usually that is why Americans have to watch 
there language when speaking and use only correct words!

Naira

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] FW: How to remove DAB to restain with DAB

[Margaryan, Naira]

Hello,

I would like to repeat my DAB staining on the same slide with DAB I run before. 
I know that acid alcohol will remove hematoxylin but how to remove DAB?
I appreciate to any suggestion.

Thanks in advance,
Naira

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] kits for LCM archival FFPE tissue

[Margaryan, Naira]

Hi all,

I am about to perform PCR on archival FFPE tissue after LCM.
My LCM system is from Arcturus and my tissue is on membraned glass slides.

My questions are:

 1.  What Kit to use to get the highest yields and quality RNA?
 2.  Can anyone who has been doing it for the past 2-3 years suggest me a good 
amplification and purification kit for FFPE tissue?
 3.  Can you please send me the serial numbers of the kits?


Thanks in advance,
Naira
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] TGFβ RII

[Margaryan, Naira]

Hi histonetters,

I need suggestion and help with the TGFβ RII (D-2): sc-17799 Ab, which should 
show a cytoplasmic staining. My problem is that I am getting a beautiful 
nuclear staining only. How I can fix this problem? What to pay attention on and 
what to change in the usual protocol? My AR is citrate buffer pH6.
This is a monoclonal mouse Ab against human TGFβ RII. This is only Ab that can 
work for me because it recognize only human TGFβ RII and does not cross react 
with mouse.

Thanks in advance,
Naira

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

RE: [Histonet] ihc slides

[Margaryan, Naira]

Hi Cynthia,

I agree with Greg, you have to re-do your staining from beginning (from AR 
step) after removing the hematoxilin.

Naira 

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] Antigen Retrieval for 10u sections in IHC

[Margaryan, Naira]

Hi tistonetters,

I have to do IHC on 10µ sections. Is procedure for Antigen Retrieval same like 
for 4-5µ (time and temperature) 

Thanks in advance,
Naira

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] RE:Tap Water or filtration system

[Margaryan, Naira]

What if we suggest to use ddH2O both for TBST preparation and in autostainers 
(before and after IHC) because the PH level is vary in different states.



Naira

**

Confidentiality Note: This message (including any attachments) is intended for 
the use of the named recipient(s) only and may contain confidential and/or 
proprietary information. If you are not the intended recipient, please contact 
the sender and delete this message. Any unauthorized use of the information 
contained in this message is prohibited.



-Original Message-

Message: 11

Date: Mon, 15 Nov 2010 10:50:50 -0700

From: Vanessa Avalos vava...@allergydermatology.com

Subject: [Histonet] Tap Water or filtration system

To: histonet@lists.utsouthwestern.edu

Message-ID: 000e01cb84ed$a49cdbd0$edd693...@com

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



We are in the process of getting our new Lecia Autostainer delivered to us.

The question came up if we will still need to continue using our water

purification system for the stainer. Mostly the docs would like to save

money but I don't want them to be unhappy with the result.



I would like to hear all the pros and cons of discontinuing/continuing the

use of our system and going w/ tap water from the sink. What is the PH level

supposed to test at? What difference will I see in the slides? By the way, I

only stain HE  at the current time but there is a possibility of starting

PAS or other special stains.



Thank you in advance!!!

V.Avalos

ADS, INC

Fax:602-277-2134
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] FW: How to remove Hematoxilin

[Margaryan, Naira]

Hi histoworld,

I would like to repeat my staining on the slides already coverslipped but need 
to remove hematoxilin first.
How to remove hematoxilin? Is it need to repeat Antigen retrieval?

Thanks in advance,
Naira

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] RE: coverslipper

[Margaryan, Naira]

Thanks a lot to all of you answered me. I was surprise nobody mentioned 
coverslipper from DAKO. Are any of you have any experience with DAKO's 
coverslipper?

Again Thanks to all,
Naira


Subject: coverslipper

Hi Colleges,

I need your opinion about coverslip by hand vs. using machine.
If you use machine what company's coverslipper you prefer?

Thanks in advance,
Naira

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

RE: [Histonet] Incubation of Ab more then 1 hour in autostainer

[Margaryan, Naira]

Dear Histonetters,

Thanks everyone for several good advices.

I'll try all suggestions: increase volume from 600ul to 800ul, add more primary 
antibody step and put hot water dishes inside of machine to keep humidity as 
high as possible.

Hopefully it will work...

Many thanks,
Naira

-Original Message-
From: Liz Chlipala [mailto:l...@premierlab.com] 
Sent: Thursday, March 18, 2010 3:05 PM
To: Margaryan, Naira; histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] Incubation of Ab more then 1 hour in autostainer

We have done up to an hour but not longer, if I were you I would have
two antibody steps and just to add some more reagent after an hour or
1.5 hours

Liz

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Manager
Premier Laboratory, LLC
PO Box 18592
Boulder, Colorado 80308
office (303) 682-3949 
fax (303) 682-9060
www.premierlab.com
 
 
Ship to Address:
1567 Skyway Drive, Unit E
Longmont, Colorado 80504

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of
Margaryan, Naira
Sent: Thursday, March 18, 2010 1:39 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Incubation of Ab more then 1 hour in autostainer
Importance: High

Hi Histonetters!

I have a stupid question but I Have to ask.

Does anyone perform an Incubation of Ab that required more then 1 hour
(2-3 hours) in autostainer? Does autostainer keep slides wet or slides
sometimes are getting dry? 

I know that slides should not be dry in any step of IHC.

Thank you very much!

Naira

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] Incubation of Ab more then 1 hour in autostainer

[Margaryan, Naira]

Hi Histonetters!

I have a stupid question but I Have to ask.

Does anyone perform an Incubation of Ab that required more then 1 hour (2-3 
hours) in autostainer? Does autostainer keep slides wet or slides sometimes are 
getting dry? 

I know that slides should not be dry in any step of IHC.

Thank you very much!

Naira

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] Bleach in autostainer cost problems?

[Margaryan, Naira]

Hi Histonetters,

Have you ever had the problem I am having now: light or totally negative 
staining after service and cleaning the machine with bleach?

To explain it better:
I have immuno-autostainer from TermoFisher that was working well before I had 
service from the company. After a representative from the company serviced the 
machine and cleaned it with bleach, my staining starts to become very light or 
totally negative on some slides which were running all together on the same 
day. Unfortunately, I did not realize it immediately and only after several 
stainings when I expect to see strong staining instead of light like 1+.

First of all, I thought it's one of the reagents or my antibody. Then, I 
checked on the positive slides and finally I came to decision that this is a 
bleach that was used to clean machine.

I can't get in touch with rep almost a week.

If you are familiar with this kind of problem, please give me any suggestion.

Naira

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] IF staining on FFPE tissue

[Margaryan, Naira]

Dear Histonetters,

Hopefully you are enjoying a great Holidays with your friends and family!

I have been asked to perform . I got positive and negative absolutely identical 
with beautiful nuclear DAPI and very red RBC. What to do or what to change in 
my protocol to avoid RBC and background? Here is my protocol:
1.Deparaffinization  (xyline- 60min, 100%Eth- 15min, 95%- 5min, 70%- 5min, H2O)
2. Antigen Retrieval ph6 in Citrate buffer (same I use for IHC)
3. Wash H2O and TBST
4. Protein block- 
10minhttps://webmail.childrensmemorial.org/owa/?ae=PreFormActiont=IPM.Notea=Replyid=RgDBaOhrx0l9S5XoN3P8OXzpBwDbYZXQd%2bZ%2fQIud7XQY1kjHAAAKzOB%2bAADBupMEDGuaSo2Eh4%2bT%2fsNqAAOO6ORIAAAJ#
5. Ab, Rb anti-human - 60-90 min (same I use for IHC)
6. Wash TBST -5-10min
7. secondary Donkey anti-Rb 594 red - 60min
8. Wash TBST -5-10min, H2O -5min
9. DAPI - 10min
10. Wash H2O -5-10min
11. Gelvatol coverslip

Am I using wrong secondary? Is there any specific secondary or protocol for IF 
staining for FFPE  to avoid background? I just used same AR and primary Ab's 
dilution like i use for IHC with nice results.

Any suggestions are appreciated, especially in these Holidays days from working 
histo- people:)

Thanks and have a warm and nice Holidays,
Naira
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] RE: IF staining on FFPE tissue

[Margaryan, Naira]

Dear Histonetters,

Hopefully you are enjoying a great Holidays with your friends and family!

I have been asked to perform . I got positive and negative absolutely identical 
with beautiful nuclear DAPI and very red RBC. What to do or what to change in 
my protocol to avoid RBC and background? Here is my protocol:
1.Deparaffinization  (xyline- 60min, 100%Eth- 15min, 95%- 5min, 70%- 5min, H2O)
2. Antigen Retrieval ph6 in Citrate buffer (same I use for IHC)
3. Wash H2O and TBST
4. Protein block- 10min
5. Ab, Rb anti-human - 60-90 min (same I use for IHC)
6. Wash TBST -5-10min
7. secondary Donkey anti-Rb 594 red - 60min
8. Wash TBST -5-10min, H2O -5min
9. DAPI - 10min
10. Wash H2O -5-10min
11. Gelvatol coverslip

Am I using wrong secondary? Is there any specific secondary or protocol for IF 
staining for FFPE  to avoid background? I just used same AR and primary Ab's 
dilution like i use for IHC with nice results.

Any suggestions are appreciated, especially in these Holidays days from working 
histo- people:)

Thanks and have a warm and nice Holidays,
Naira

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] anti-human Notch4

[Margaryan, Naira]

Hi friends,

My PI is asking for Notch 4 IHC on FFPE tissue. Can any of you suggest me a 
good Ab (preferably mot mouse but anti-human Notch4) and protocol please!

Thanks in advance,
Naira

Naira V. Margaryan, D.V.M., Ph.D.
Research Scientist
Children's Memorial Research Center
2300 Children's Plaza, Box 222
Chicago, IL 60614-3363
Tel: 773-755-6340
Fax: 773-755-6594
nmargar...@childrensmemorial.orgmailto:nmargar...@childrensmemorial.org

For Express Mail:
CMRC, Room C.473
2430 N. Halsted Street
Chicago, IL  60614-4314

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] AlkPhos

[Margaryan, Naira]

Dear histonetters,

I have to order AlkPhos kit to my IHC and, because I did not use it for the 
past 5-6 years, I do not know what company will best to order from.

Any suggestions are appreciated,

Naira

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] human macrophages in human melanoma xenografts

[Margaryan, Naira]

Hi Histonetters,

I would like to be able to look at human macrophages in human melanoma 
xenografts raised in mouse. Could you please suggest me a best Ab for ICH and 
protocol?



Thanks in advance,

Naira


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

RE: [Histonet] human macrophages in human melanoma xenografts

[Margaryan, Naira]

Thanks a lot both of you!

For melanoma detection, I usually use HRP with AEC. Is there any non-mouse 
anti-human macrophage marker?

Thank you much,
Naira 


-Original Message-
From: Liz Chlipala [mailto:l...@premierlab.com] 
Sent: Friday, October 23, 2009 10:51 AM
To: Galbraith, Joe; Margaryan, Naira; histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] human macrophages in human melanoma xenografts 

F4/80 is a mouse macrophage marker.  If she wants to detect human
macrophages in a mouse background she will need to use a mouse
anti-human CD68.  She will need to run it with a mouse on mouse
detection system and run all of the appropriate negative controls.  I
would also select a alkaline phosphatase detection system rather than
HRP with DAB just incase the tumor has any melanin in it.  I have done
this before looking for human lymphocytes with LCA in a mouse
background.  You just need to make sure you have all of the appropriate
controls.  For a positive control I would use a human tonsil or
something like that and you need to make sure you run the appropriate
isotype negative controls.  

Liz

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Manager
Premier Laboratory, LLC
PO Box 18592
Boulder, Colorado 80308
office (303) 682-3949 
fax (303) 682-9060
www.premierlab.com
 
 
Ship to Address:
1567 Skyway Drive, Unit E
Longmont, Colorado 80504

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of
Galbraith, Joe
Sent: Friday, October 23, 2009 9:44 AM
To: Margaryan, Naira; histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] human macrophages in human melanoma xenografts 

Naira:

Here is a link to a site listing macrophage markers.  F4/80 is a
commonly mentioned marker for macrophages.  I presume you mean IHC
rather than ICH.  Enjoy.

http://www.antibodybeyond.com/reviews/cell-markers/macrophage-marker.htm

Joe
joseph-galbra...@uiowa.edu


-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of
Margaryan, Naira
Sent: Friday, October 23, 2009 10:31 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] human macrophages in human melanoma xenografts 

Hi Histonetters,

I would like to be able to look at human macrophages in human melanoma
xenografts raised in mouse. Could you please suggest me a best Ab for
ICH and protocol?



Thanks in advance,

Naira


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] Isotype background

[Margaryan, Naira]

Hi Adam,

I always do the isotype control parallel with a no-primary control in parallel 
with my real Ab. 

May I suggest you to use Protein block serum free and pure Ab diluent without 
adding 2% donkey serum?

Try this and let me know your results.

If histonetters think I am wrong, fill free and please, let me know. 

All the best,
Naira 

-Original Message--

Message: 11
Date: Tue, 13 Oct 2009 16:48:46 -0500
From: Adam . anonwu...@gmail.com
Subject: [Histonet] Isotype background
To: histonet@lists.utsouthwestern.edu
Message-ID:
858249120910131448v26622f8dtd5599dbbf64e6...@mail.gmail.com
Content-Type: text/plain; charset=ISO-8859-1

Hi all,

I am trying some IHC, and I am having a peculiar problem. Like I expect, my
antibody of interest (anti-mouse goat polyclonal) stains nonspecifically at
high concentrations (10 ug / ml) but as I titer it down (3 ug / mL), it
seems to stain relatively specifically the cells I think it should stain.
However, at 3 ug / mL, my isotype goat IgG stains nearly everything.

Here is my protocol
1) Block in 3% H2O2 for 10'. Wash.
2) Block in 10% donkey serum for 1 hr. Wash.
3) Block in avidin 15', wash, biotin 15', wash (Vector Labs blocking kits)
4) Incubate with primary / isotype overnight at 4C in 2% donkey serum. Wash.
5) Incubate with secondary (biotinylated donkey anti-goat, cross adsorbed to
mouse) in 2% donkey serum for 1 hr at room temp. Wash.
6) Incubate with strepavidin HRP in TBS-T for 30'. Wash.
7) Incubate with DAB+ (Dako) for 5'.

For isotype, I am using Chrompure Goat IgG, whole molecule from Jackson.
Some people have suggested that I just do away with isotypes altogether and
use a no primary control instead. I think there is some merit to this idea,
but I still think my issue might be indicative of a larger technical problem
in my staining protocol.

Thanks,
Adam

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] black DAB instead of brown

[Margaryan, Naira]

Dear Histometters:

I am getting black staining with my DAB instead of brown.
I use usual protocol with AR-pH6, H2O2, Avidin/Biotin, PB, 1º, biotinylated 
2-dary, streptavidin then DAB.

Does any of use know this kind of problem? and What exactly can coast this 
artifact?

Thanks in advance,
Naira

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] acid phosphatase in tissues in routine IHC

[Margaryan, Naira]

Hi  histonetters,

Around a month ago, I asked a question and did not get any answer, but I really 
would like to have any thoughts from you about the: Would the presence of acid 
phosphatase in tissues cause non-specific background staining when doing 
routine immunoperoxidase staining -- DAB? If answer is YES: how to block it?

Thanks in advance,
Naira
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] type of paraffin and polymer

[Margaryan, Naira]

Dear Histonetters,

I quite need answer as soon as it possible, PLEASE!

I am working with mouse tissue and tumors. What type of paraffin is the best 
for processing and embedding to cut 4µ nice sections?

Thanks in advance,
Naira

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] RE: reagents for IHC

[Margaryan, Naira]

Hi Everyone,

I do appreciate for all suggestions I get form most of you!

Have a nice week,

Naira

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] reagents for IHC

[Margaryan, Naira]

Hi everyone,

How are you experiencing the economic pressures and price changes for REAGENTS? 
I am sorry, but I just bought reagents from DAKO and, for the price I paid for 
125 ml before, I got 15-50 ml :(

I am ready to switch to another company, but I need your suggestion about 
reagents for IHC and companies you are purchasing from: Peroxidase Block, 
Protein Block, Antibody diluents, DAB, AEC, different secondaries and 
tertiaries.

Thanks in advance,
Naira

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] bone fixation and calcification

[Margaryan, Naira]

Dear Histonetters,

I am new in fixation and processing bones. I need your full protocol with 
details how to fix and process mice bone to visualize the tumor metastases?

Thanks in advance,
Naira
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] Fixation of Formalin vs Bouin's for IHC

[Margaryan, Naira]

Hi histonetters,


Please help. What is difference in procedure for IHC between 10%Formalin fixed 
tissue and in Bouin's fixed tissue? I used to use 10%FFPE tissue with Citrate 
buffer Antigen Retrieval for my IHC. Do I have to change my protocol for the 
Bouin's fixed tissue?

Hope for you soon response and thanks in advance,
Naira

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] Trypsin versus other AR-s

[Margaryan, Naira]

Hi Dears,

 

It is Friday, but not a weekend yet:-):-(

 

I just got a request from my PI to figure out:

1.  How often now day's people use Trypsin (EDTA, Proteinase K or E)
as an Antigen Retrieval for FFPE. 

2.  Why or is the Citrate Buffer pH6 more usable??? 

3.  Is Trypsin very old technique? 

 

Any educational feedback is appreciated,

Naira

 

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] keratin 18

[Margaryan, Naira]

Hi histoneters,

 

Please help me to find an Ab to detect keratin 18 in mouse tissue, this
means that at has to be not mouse Ab.

 

Thanks in advance,

Naira

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] chicks' embryos and LCM

[Margaryan, Naira]

Dear histoneters,

 

I am going to start the LCM work on chicks' embryos and now more
questions are coming up :-)

1.What kind of brand of blades will you suggest to use for
crysectioning (Catalog ## will be very good)?

2.After placing glass slides in LCM what kind of caps will you
suggest using to capture peaces: CupSureHS or Macro?

3.What is minimum amount of peaces I have to capture to get good
quality of RNA?

 

 

Thanks in advance,

Naira

 

Naira V. Margaryan, D.V.M., Ph.D.

Research Scientist

Children's Memorial Research Center

2300 Children's Plaza, Box 222

Chicago, IL 60614-3363

Tel: 773-755-6340

Fax: 773-755-6594

nmargar...@childrensmemorial.org
mailto:nmargar...@childrensmemorial.org 

 

For Express Mail: 

CMRC, Room C.473

2430 N. Halsted Street

Chicago, IL  60614-4314

 

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] RE: HE help (Weems, Joyce)

[Margaryan, Naira]

Hi Roger,

I usually use Bluing reagent (Richard Allan Scientific, part of Fisher)
after Hematoxylin and tap water, then wash again before eosin. Try it
and you will see big difference. I just love it!

All the best,
Naira
--

Message: 1
Date: Sat, 10 Jan 2009 13:33:09 -0500
From: Weems, Joyce jwe...@sjha.org
Subject: RE: [Histonet] HE help
To: lpw...@sbcglobal.net, Charles, Roger rchar...@state.pa.us,
Histonet histonet@lists.utsouthwestern.edu
Message-ID:

5d64396a0d4a5346bebc759022aaeaa5257...@itsssxm01v6.one.ads.che.org
Content-Type: text/plain; charset=us-ascii

Also, warm water works best for bluing, if you have that option..

Joyce Weems 
Pathology Manager 
Saint Joseph's Hospital 
5665 Peachtree Dunwoody Rd NE 
Atlanta, GA 30342 
678-843-7376 - Phone 
678-843-7831 - Fax 

 

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Lee 
Peggy Wenk
Sent: Saturday, January 10, 2009 11:00 AM
To: 'Charles, Roger'; 'Histonet'
Subject: RE: [Histonet] HE help

It might not be a hematoxylin problem. It might be too much eosin.

Try one of the following:
- cut the eosin time down to 1 minute, and if it's still too pink/not
blue enough, cut the eosin time down to 30 seconds.
- and/or, change the first 95% alcohol after the eosin to 70% alcohol.
That will help to put out more eosin than 95%. 

Peggy A. Wenk, HTL(ASCP)SLS
Beaumont Hospital
Royal Oak, MI 48073

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Charles,
Roger
Sent: Friday, January 09, 2009 9:14 AM
To: Histonet (histonet@lists.utsouthwestern.edu)
Subject: [Histonet] HE help

Hello All,
TGIF
I'm trying to help out one of my veterinarian pathologist in getting
some information on how to increase the bluing of the HE staining.
Presently we are using pre made Mayers Hematoxylin from Sigma with an
automated schedule as follows:

 1.  Drying 45min
 2.  citrisolve 3 min
 3.  citrisolve 30 sec
 4.  citrisolve 3min
 5.  100% 45 sec
 6.  100% 45 sec
 7.  100% 3 min
 8.  95% 30 sec
 9.  95% 1 min 30 sec
 10. Tap water 1 min
 11. Hematoxylin 8 min
 12. tap water 4 min
 13. eosin 1 min 30 sec
 14. 95% 1 min
 15. 95% 1 min
 16. 95% 1 min
 17. 100%1 min
 18. 100%1 min
 19. 100%1 min
 20. Citrisolve 1 min
 21. Citrisolve 1 min
 22. Citrisolve 1 min
 23. Citrisolve up to 120 min
The pathologist is stating the slides are too eosinic and would like
greater bluing.  My question is are there limitations to the Mayers
Hematoxylin on how blue it will actually get and is there something else
we can change in our schedule to increase the bluing or decrease the
eosin to get the desired affect?
Thanks to all


Roger Charles
Microbiologist
Pennsylvania Veterinary Laboratory
2305 N Cameron St
Harrisburg, PA 17110
717-787-8808

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Confidentiality Notice:
This email, including any attachments is the 
property of Catholic Health East and is intended 
for the sole use of the intended recipient(s).  
It may contain information that is privileged and 
confidential.  Any unauthorized review, use,
disclosure, or distribution is prohibited. If you are 
not the intended recipient, please reply to the 
sender that you have received the message in 
error, then delete this message.




--

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

End of Histonet Digest, Vol 62, Issue 13


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet