[Histonet] Job opening La Jolla Ca

[dusko trajkovic via Histonet]

Core histoligy job in La Jolla CA, day shift, for a well know academic 
institution.
If interested, please contact me for more info.
Dusko Trajkovic

Sent from my iPhone

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Re: [Histonet] IHC and Histogel

[dusko trajkovic via Histonet]

We have used Histogel for years on cell pellets, and have not come across any 
IHC issues.Dusko 

On Thursday, May 10, 2018 7:13 AM, Paula via Histonet 
 wrote:
 

 Hello good morning everyone,

Can someone reply who knows if Histogel causes any interference with IHC
staining?

Thank you in advance,

Paula

Bio-Path Med Group

Fountain Valley, CA

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Re: [Histonet] Mouse bladder

[dusko trajkovic via Histonet]

I agree with Colleen, processing is the issue. You need to process on a longer 
program in order to dehydrate and clear the agar properly.dusko 

On Tuesday, December 19, 2017 9:45 AM, Colleen Forster via Histonet 
 wrote:
 

 Be sure you are processing them on an overnight process. Even if the
samples are small it takes that time to process the agar properly.

I use Histogel and learned this lesson the hard way. My samples always
"shriveled" up too until someone enlightened me about the processing time.
I would think agar might be the same.


Just a thought,

Colleen Forster HT(ASCP)QIHC
U of MN

On Tue, Dec 19, 2017 at 11:29 AM, Liz Chlipala via Histonet <
histonet@lists.utsouthwestern.edu> wrote:

> I would try histogel instead of agar.
>
> Liz
>
> Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
> Premier Laboratory, LLC
> PO Box 18592
> Boulder, CO 80308
> (303) 682-3949 office
> (303) 682-9060 fax
> (303) 881-0763 cell
> l...@premierlab.com
> www.premierlab.com
>
> Ship to Address:
>
> Premier Laboratory, LLC
> 1567 Skyway Drive, Unit E
> Longmont, CO 80504
>
> From: Heather Marlatt via Histonet [mailto:histonet@lists.
> utsouthwestern.edu]
> Sent: Tuesday, December 19, 2017 9:12 AM
> To: histonet@lists.utsouthwestern.edu
> Subject: [Histonet] Mouse bladder
>
> Does anyone have a good protocol for mouse bladder embedded in agar that
> they are willing to share? We have mouse bladders in 2% agar and have tried
> a few different protocol variations from our usual mouse but the agar keeps
> shriveling up.
>
> Thanks
> Heather
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> Histonet@lists.utsouthwestern.edu >
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>
> 
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>
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[Histonet] Histo job opportunity in a GI lab-Oak Lawn Illionios

[dusko trajkovic via Histonet]

If anyone is interested in a histology position in a GI lab, please let me know 
as soon as possible?Lab is located in Oak Lawn Illinois.At the moment I do not 
have details, but will be getting them soon.ThanksDusko Trajkovic760-390-3478
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[Histonet] CD16/FCGR4 Antibody-Mouse specific

[dusko trajkovic via Histonet]

Hello Histonetters,Does anyone have any experience with this antibody? Any 
information, pos or neg, would be greatly appreciated.Have a good 
day.thanksDusko Trajkovic
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[Histonet] Job Opportunity in So California

[dusko trajkovic via Histonet]

Hallo Histonetters,

We have a great job opportunity at Pfizer La Jolla site, inthe Investigative 
Pathology Lab.

Here is a short list of qualifications desired. Emphasis onIF/IHC multiplexing.

 If you can fulfilthese, please contact me via email (dunat...@sbcglobal.net) 
and I will provide you with information on how to apply through the 
properchannels.

This Pfizer site is nestled in one of the most beautifulplaces in Southern 
California, close to Torrey Pines and UCSD. 


Development and conduct of advancedImmunohistochemistry/Immunofluorescence 
procedures on preclinical study samplesin support of the rapidly expanding 
field of immuno-oncology.  Role toserve as a technical expert for the 
discipline and for projects supported. 
• Responsibilities: 
• Technical support of current Immunohistochemistry/Immunofluorescence 
workloadwith primary focus on immuno-oncology projects. 
• Identification of Immunohistochemistry/Immunofluorescence assay/analysisgaps, 
development of work solutions 
• Single plex and multi-plex  Immunohistochemistry/Immunofluorescencemethod 
development/optimization/validation 
• Technical expert for discipline and appropriate project teams/research 
unitworking groups 
• Immunohistochemistry/Immunofluorescence process (simple/complex) mentor 
forjunior colleagues 
• Development Expectation: 
• Image Analysis (chromogenic or fluorescence) – including data 
interpretation,integration and presentation 
• More in depth project team involvement – Drug Safety Team Lead

 Dusko Trajkovic

Scientist-Investigative path Lab

Pfizer Inc, La Jolla

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[Histonet] alpha V beta 3 antibody search

[dusko trajkovic via Histonet]

Hi Everyone, I have tried 2 different aVb3 antibodies, and they do not seem to 
work, or rather stain properly. Does anyone have a aVb3 antibody that will work 
on FFPE sections? Any info would be greatly appreciated.ThanksDusko
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[Histonet] Looking for a used tissue processor

[dusko trajkovic]

I'm in a market for a used tissue processor, preferably a  Sakura VIP 5. I am 
located in San Diego area.Please contact me at  dunat...@sbcglobal.net
858-638-6202ThanksDusko
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[Histonet] MUC1 antibody inquiry

[dusko trajkovic]

Hallo Everyone,
I am looking for information on MUC1 antibody that is known to react only to 
Human, and does not cross-react with the mouse for FFPE IHC procedure. Any 
suggestions/advice will be greatly appreciated.
Thank you and have a good Monday!!
Dusko
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[Histonet] Chlamydia infected tissue

[dusko trajkovic]

Hi,
Does anyone have chlamydia infected tissue, or info on where I might possibly 
get some? I need to optimize this antibody, but do not have positive control 
tissue. 
Any info would be greatly appreciated.
thank you
Dusko Trajkovic
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Re: [Histonet] Histogel

[dusko trajkovic]

Jennifer,
You might have seen one of my posts from 2-3 years ago. I had the exact same 
problems you described. Could not get anyone to come up with a solution. I ran 
various programs on our VIP and finally came up with a solution. 
Fix your specimens as you normally would do. Drain of the fixative add your 
histogel (dissolved in hot water, which you have been doing), fill the mold 
with the histogel. Let solidify on ice or 4C in fridge. Place the solid 
histogel in a cassette and process on a 12 hour program. Since I have 
instituted this procedure, have not had one bad block to date. Longer 
processing is the answer, and nothing else.
Good Luck.
Dusko Trajkovic 
Pfizer Inc. La Jolla
858-638-6202
 


 From: jennifer.arcand-john...@genzyme.com 
jennifer.arcand-john...@genzyme.com
To: histonet@lists.utsouthwestern.edu 
Sent: Monday, January 20, 2014 7:56 AM
Subject: [Histonet] Histogel
  

Dear Histonetters,

I have been reading up on the archives for info on Histogel.  Previous posts 
discuss how they had problems with it - some samples would come out great and 
some would shrivel up or even dissolve.
These posts on the Histogel were from a few years ago and was hoping, and 
praying that someone out there may have solved this issue and have a little 
info you could share with me on this subject.  Did anyone out there ever figure 
out how to get consistent results?
I have spoken with RA Scientific and they have no additional insights.

Here is the background:  I have used Histogel for about 4 years now.  In 
September of last year, we started seeing the shriveling Histogel samples.  
Like others who posted, it was random.  I could embed two serial pieces of 
nerve, from the same mouse, into two blocks and one would shrivel and one would 
look great.  So I have tried many things, always in multiples of 3 or more per 
condition per run...

Fixing in formalin only, embedding in Histogel and storing in PBS until 
processing
Fixing in formalin only, embedding in Histogel and storing in 40% reagent 
alcohol until processing (the first step of our processor is 40%)
Fixing in formalin only, embedding in Histogel and storing in formalin until 
processing
Fixing in formalin, rinsing in 40% alcohol, embedding in Histogel, and storing 
in 40% or formalin until processing
For all of these conditions, I have tried using a small cycle (30 min/bath) and 
a biopsy cycle (15 minutes/bath).
Once processed, there was no rhyme or reason to the results.  Some blocks 
looked great; others within the same group looked shriveled.  Sometimes the 
blocks were white, sometimes they were clear.

Next, I thought it was my pre-processing - so I heated the Histogel in a water 
bath, rather than microwaving.  That way all of the samples were embedded with 
the Histogel at the same temperature - about 55 degrees.
Again, no rhyme or reason, some looked good, some looked bad.

Lastly I thought that maybe I was carrying over too much liquid from my sample 
to the Histogel so I tried the following:
I made Histogel only blocks or added 3-4 drops of 40% alcohol or formalin to 
the liquid Histogel before a tissue/cell free block was made.
Yep, you guessed it - no luck.  Some looked good, some looked shriveled.

So here I have this great tool to embed tiny samples, but I am afraid to use it 
because I don't know if it will work or shrivel!  Can anyone out there help me?
Thanks,
Jenn


Jennifer Johnson

Staff Scientist

Genzyme, a Sanofi Company

Department of Pathology

5 Mountain Road

Framingham, MA 01701-9322

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Re: [Histonet] Histogel

[dusko trajkovic]

Esther,
I mainly process cells, which have been spun down into a small pellet. Also 
mouse DRG's and other very small tissues. I would consider this delicate, so do 
not be afraid to use a longer processing program. Histogel/Agurose is what 
needs longer dehydrating steps. 
We do not use any substitute reagents, so in that aspect I cannot tell you how 
they will affect the processing. Our lab uses ethanol, xylene, and Paraplast 
paraffin. Try a test run and let me know if you were able to get successful 
results. 
Have a good Monday!
Dusko
 


 From: Esther C Peters epete...@gmu.edu
To: jennifer.arcand-john...@genzyme.com 
jennifer.arcand-john...@genzyme.com; histonet@lists.utsouthwestern.edu 
histonet@lists.utsouthwestern.edu; dusko trajkovic dunat...@sbcglobal.net 
Sent: Monday, January 20, 2014 11:15 AM
Subject: RE: [Histonet] Histogel
  

Thank you, Dusko!

I have had the same problem with 1.5% agarose, and I tried starting the 
dehydration with 30% to 50% to 70% ethanol, and using different xylene 
substitutes. It appears that the variable whitening and shrinking happens after 
100% reagent alcohol and in the xylene substitute (now using Richard-Allan 
Clear-Rite3). I've wondered if slow infiltration was the issue.  I guess we'll 
try this longer processing, but I also work with delicate tissues that normally 
would be a short run (15 min in each reagent). Are your tissues 
thin/delicate/biopsy or cell preps or organ samples? No effect on them?

Esther

Esther C. Peters, Ph.D.
Assistant Professor
Environmental Science  Policy
George Mason University
4400 University Drive, MS 5F2
Fairfax, VA 22030-

From: histonet-boun...@lists.utsouthwestern.edu 
histonet-boun...@lists.utsouthwestern.edu on behalf of dusko trajkovic 
dunat...@sbcglobal.net
Sent: Monday, January 20, 2014 1:58 PM
To: jennifer.arcand-john...@genzyme.com; histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] Histogel

Jennifer,
You might have seen one of my posts from 2-3 years ago. I had the exact same 
problems you described. Could not get anyone to come up with a solution. I ran 
various programs on our VIP and finally came up with a solution.
Fix your specimens as you normally would do. Drain of the fixative add your 
histogel (dissolved in hot water, which you have been doing), fill the mold 
with the histogel. Let solidify on ice or 4C in fridge. Place the solid 
histogel in a cassette and process on a 12 hour program. Since I have 
instituted this procedure, have not had one bad block to date. Longer 
processing is the answer, and nothing else.
Good Luck.
Dusko Trajkovic
Pfizer Inc. La Jolla
858-638-6202



From: jennifer.arcand-john...@genzyme.com 
jennifer.arcand-john...@genzyme.com
To: histonet@lists.utsouthwestern.edu
Sent: Monday, January 20, 2014 7:56 AM
Subject: [Histonet] Histogel


Dear Histonetters,

I have been reading up on the archives for info on Histogel.  Previous posts 
discuss how they had problems with it - some samples would come out great and 
some would shrivel up or even dissolve.
These posts on the Histogel were from a few years ago and was hoping, and 
praying that someone out there may have solved this issue and have a little 
info you could share with me on this subject.  Did anyone out there ever figure 
out how to get consistent results?
I have spoken with RA Scientific and they have no additional insights.

Here is the background:  I have used Histogel for about 4 years now.  In 
September of last year, we started seeing the shriveling Histogel samples.  
Like others who posted, it was random.  I could embed two serial pieces of 
nerve, from the same mouse, into two blocks and one would shrivel and one would 
look great.  So I have tried many things, always in multiples of 3 or more per 
condition per run...

Fixing in formalin only, embedding in Histogel and storing in PBS until 
processing
Fixing in formalin only, embedding in Histogel and storing in 40% reagent 
alcohol until processing (the first step of our processor is 40%)
Fixing in formalin only, embedding in Histogel and storing in formalin until 
processing
Fixing in formalin, rinsing in 40% alcohol, embedding in Histogel, and storing 
in 40% or formalin until processing
For all of these conditions, I have tried using a small cycle (30 min/bath) and 
a biopsy cycle (15 minutes/bath).
Once processed, there was no rhyme or reason to the results.  Some blocks 
looked great; others within the same group looked shriveled.  Sometimes the 
blocks were white, sometimes they were clear.

Next, I thought it was my pre-processing - so I heated the Histogel in a water 
bath, rather than microwaving.  That way all of the samples were embedded with 
the Histogel at the same temperature - about 55 degrees.
Again, no rhyme or reason, some looked good, some looked bad.

Lastly I thought that maybe I was carrying over too much liquid from my sample

Re: [Histonet] Unsubscribe, Chapter 195

[dusko trajkovic]

Peggy,
I don't think anyone else could have said it better.
Thank you
Dusko



From: Lee  Peggy Wenk lpw...@sbcglobal.net
To: Manfre, Philip philip_man...@merck.com; nmhi...@comcast.net; HISTONET 
histonet@lists.utsouthwestern.edu 
Sent: Monday, September 9, 2013 7:47 AM
Subject: Re: [Histonet] Unsubscribe, Chapter 195


I'm going to wade in, not as someone who has posted numerous times on how to 
unsubscribe, but as someone assessing it from a risk assessment evaluation.

If there is a lab task that is consistently being done wrong, by many different 
people, it is usually NOT the fault of the people. It is either a training 
issue, or a process problem. So we either have to do a better job training and 
re-training, or we need to change how the process/procedure is being done.

With the Histonet email, since people are constantly joining, often for a day 
or two, we can't really improve the training aspect. Yes, there are 
instructions when we first join, to print off/save how to subscribe or 
unsubscribe or change personal information, etc. But (be honest) how many of us 
pay attention to these types of instructions when we sign up to be a member 
of a credit card or a on-line department store or an on-line book store or 
other email lists? Most people do not. So we know that this type of training 
is not effective. But we really can't do a one-on-one type of training session 
for each person who signs on to Histonet. Therefore, improving the training is 
not the answer.

The answer lies in modifying the process. Look at the bottom of those emails 
from credit cards or hotels or department stores that you have signed up with. 
There is usually a line that says If you no longer wish to receive these 
emails, click on this link and follow the instructions.

Add to that, various email lists have various methods on how to unsubscribe, 
which can involve a link, or putting the word unsubscribe in the subject, or 
putting the word unsubscribe in the message.

Histonet has a link at the bottom, but no instructions. So it's not clear to 
click on the link to unsubscribe, nor is there any mention whether one of the 
other unsubscribing methods would work. I therefore believe the Histonet 
unsubscribing procedure has a process problem, that could be easily fixed.

As for the fact that how to unsubscribe has been explained 5,391+ times in the 
past does not help the person who signed up over the weekend, and as of today, 
decided that Histonet is not what they need. This new person has not seen the 
previous requests for help with unsubscribing, nor the answers on how to do it. 
Again, this is a process problem.

Is there any way Histonet can get some clearer instructions at the bottom of 
each email, on how to unsubscribe, either permanently or temporarily while on 
vacation? Such as saying To unsubscribe, click on the link below, and follow 
the instructions at the bottom of the next webpage.

Let's not yell at the people trying to unsubscribe. Let's work on improving the 
unsubscribing process, so we don't get these requests.

Peggy A. Wenk, HTL(ASCP)SLS

-Original Message- From: Manfre, Philip
Sent: Friday, September 06, 2013 7:30 AM
To: nmhi...@comcast.net ; HISTONET
Subject: RE: [Histonet] Unsubscribe, Chapter 195

Trained professionals should know by now that if you want to unsubscribe, you 
must type in all caps - UNSUBSCRIBE


Philip Manfre, B.A., HT (ASCP)
Associate Principal Scientist
Merck Research Laboratories
WP45-251
PO Box 4
West Point, PA 19486

215-652-9750
215-993-0383 (fax)
philip_man...@merck.com




-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of 
nmhi...@comcast.net
Sent: Thursday, September 05, 2013 9:57 PM
To: HISTONET
Subject: [Histonet] Unsubscribe, Chapter 195

It is a concern that members of our  technically-oriented career field  have a 
difficult time understanding the method for unsubscribing to Histonet. There is 
an  almost- daily posting to unsubscribe, despite the fact that this subject 
has been addressed literally hundreds of times.  When one joins Histonet, 
instructions are provided, should be printed out for reference and used if the 
subscriber decides to leave the group.  We are required to be knowledgeable on 
all manner of technical routines requiring detailed instructions and  Histonet 
is no less clear in the methods for joining and un-joining.  Use them, 
please.  Fire away - I'm retired and I can take the flak!  I do miss my 
microtome, though...
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New Jersey, USA 08889), and/or its affiliates Direct contact information
for affiliates is 

[Histonet] Cryostat Repair Service in San Diego

[dusko trajkovic]

Good Morning,
Can anyone recomend a good, reliable cryostat repair service in the San diego 
area?
thanks
Dusko
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[Histonet] Fw: PSCA Primary antibody

[dusko trajkovic]

Hallo Histonetters,
Happy New Year!!
I'm looking for information on a PSCA (Prostate Specific Cell antigen) antibody 
that will work both in human and primate.
If are are using this antibody, please contact me. Any information would be 
appreciated.
thanks
Dusko Trajkovic
858-638-6202
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[Histonet] PSCA Primary antibody

[dusko trajkovic]

Hallo Histonetters,
Happy New Year!!
I'm looking for information on a PSCA (Prostate Specific Cell antigen) antibody 
that will work both in human and primate.
If are are using this antibody, please contact me. Any information would be 
appreciated.
thanks
Dusko Trajkovic
858-638-6202
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[Histonet] Looking for a working microtome

[dusko trajkovic]

Hallo Histo colleagues,
I'm posting this for a friend of mine who is looking for a working microtome. 
He 
works in an academic lab (San Diego area) and not a lot of money available. 
Replies from various vendors are welcome as well as individual labs looking to 
get rid of some of their unused microtomes.
thank you and have a good Wednesday
Dusko
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[Histonet] CSH 2012 Symposium-San Diego CA

[dusko trajkovic]

“Come Ride the Waves of Innovation”
 
This year’s meeting will be at the Bahia resort hotel on Mission Bay.  2 Blocks 
from the Ocean.  We are Presently lining up an outstanding list of lecturers.  
Classes will be held on 2 paddle wheel boats.  Take a look at the hotel website 
and start planning now.  Details and registration will be posted soon on the 
Society website.
 
http://www.bahiahotel.com/
http://www.californiahistology.org/events.html
 
The 2012 Symposium/Convention will be held in May in San Diego, CA.
When: May 3 - 6, 2012
Hotel Information:
Bahia Resort Hotel
998 West Mission Bay Drive, San Diego CA 858.488.0551 Reserve your room now at: 
https://shop.evanshotels.com/bahia_groups/casocf1205093.html
 
Note: To receive the group rate of $109.00 per night please mention CSH when 
registering with the hotel. The cutoff date for the group rate is April 3, 
2012.  We have already met our room number minimum, but more rooms are 
available 
at the discount rate.  Register and book you room soon.
 
We still have a few vendor tables available, but they are filling fast.
 
 
James Watson HT  ASCP
GNF  Genomics Institute of the Novartis Research Foundation
Tel    858-332-4647
Fax   858-812-1915
jwat...@gnf.org
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[Histonet] 2012 California Society for Histotechnology Symposium in San Diego

[dusko trajkovic]

Subject: CSH Meeting May 3-6
 
This year’s meeting will be at the Bahia resort hotel on Mission Bay.  2 Blocks 
from the Ocean.  We are Presently lining up an outstanding list of lecturers.  
Classes will be held on 2 paddle wheel boats.  Take a look at the hotel website 
and start planning now.  Details and registration will be posted soon on the 
Society website.  

 
http://www.bahiahotel.com/
http://www.californiahistology.org/events.html
 
 
The 2012 Symposium/Convention will be held in May in San Diego, CA. 
When: May 3 - 6, 2012
Hotel Information:
Bahia Resort Hotel
998 West Mission Bay Drive, San Diego CA 858.488.0551 
Reserve your room now at: 
https://shop.evanshotels.com/bahia_groups/casocf1205093.html
 
Note: To receive the group rate of $109.00 per night please mention CSH when 
registering with the hotel. The cutoff date for the group rate is April 3, 
2012. 

 
 
 
  
James Watson HT  ASCP
GNF  Genomics Institute of the Novartis Research Foundation
Tel    858-332-4647
Fax   858-812-1915
jwat...@gnf.org
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[Histonet] Need a protocol for CD24 and CD133

[dusko trajkovic]

Hi.
Does anyone out there have a protocol for CD24 and CD133 that will work on 
mouse 
Xenograft tumors? Preferably a protocol that will work on Leica Bond, but any 
working protocol will do at this point.
Thank you and have a good day.
Dusko
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Re: [Histonet] Histogel Problem

[dusko trajkovic]

Amos,
I have had the same problem in the past, and posted my issues on the Histonet, 
however no one was able to help me out. At one point I even exchanged Histogel 
with a colleage severl hundred miles away, thinking that maybe I had a bad lot 
of Histogel. She did not have a problem with my Histogel and neither did I with 
her Histogel. The shrinking was arbitrary. At times all of them look great and 
other times more than half shrunk and looked brittle. Even RA/Thermo did not 
have an answer for me.
I decided to do an experiment. To not bore everyone on the Histonet and explain 
all of my experimental steps, what it boiled down to is that you need a long 
processing program on the processor. We use a VIP processor as well, and the 
processing program is at least 12 hours long. NO MORE PROBLEMS. 

Since I started using this progrem, every single Histogel block has been 
perfect.
Let me know if you need any further info or explanation.

Dusko Trajkovic HT ASCP
Pfizer Inc. La Jolla

 




From: Amos Brooks amosbro...@gmail.com
To: histonet@lists.utsouthwestern.edu
Sent: Fri, June 24, 2011 10:29:09 AM
Subject: [Histonet] Histogel Problem

Hi,
  I have a problem with some blocks that were prepared with Histogel. I was
hoping someone else might have had a similar problem and figured it out. I
took a photo of the blocks that were mads and put them in a Picassa album
here:
https://picasaweb.google.com/lh/photo/APO3HsIMa2_jPOEs3QRGUjhz3qi22FNPb2i5JJnBCAk?feat=directlink

  The long  short of it is that the blocs were prepared by the researcher
for me to process. They are mouse kidneys. Now it is entirely possible for
him to have goofed something up in preparing the Histogel blocks. I wasn't
there when he did it, but when I looked at them before processing, they all
looked fine. (Like the adjacent good one in the photo). When they were
processed they were placed in the VIP right next to each other. When I went
to embed them this morning all but one of the four looked fine. The one that
didn't come out well looked like the Histogel had shrunk up and shriveled
around the kidneys. I am sure this will be aweful to cut, and the researcher
is going to have a bird over it since this happened with another project
previously. I would like to have a decent explanation for him, so if anyone
knows what might have happened and has suggestions I would love to hear it
(yes vendors too are welcome to answer this of course!).
  By the way, this was processed on a rather short cycle of 15-20 min per
station of graded ETOH from 70% to 100% with 3 xylene stations and 4
paraffin stations (45 min for these). It seems fine for everything else that
was on the processor. Just that one Histogel block was the issue.

Thank you,
Amos
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[Histonet] Temporary (8 months) Histology position in San Diego

[dusko trajkovic]

Hi Everyone,
There is a position for a  Histotechnologist at one of the major Pharmaceutical 
companies located in the Torrey Pines/SDSU/La Jolla area.
Here is the job description and if anyone knows of someone that is interested 
please have them contact me.
 
Thank you
 
Dusko Trajkovic
858-638-6202
 
Temporary Histology position:
 
Position will be for approximately 8 months.  It is approved for 40 hours per 
week but there is some flexibility and as long as there is overlap with a lead 
histotech the time of day could be flexed.  It is an  associate scientist III 
level and will be competitive.  This would be a good opportunity for someone 
who 
may want to see what industry is like and to also get a crash course in IHC as 
we do a lot here.  It is all non-GLP.  

 
Specific responsibilities in assisting the histology lab may include:
 
 
 
· Trim formalin fixed rat/mouse organs
 
· Process and embed tissue specimens
 
· Cut frozen  and paraffin (mainly) sections
 
· Perform HE stains and special stains as needed
 
· Perform IHC staining manually or by autostainer
 
· Label cassettes and slides
 
· Change reagents in tissue processer and autostainer
 
· Maintain tissue blocks and slide inventories
 
 
 
 
 
Requirements:
 
·    BS in a scientific discipline
 
·    5 years in histology
 
·    Tissue processor (VIP), coverslipper, microtome, cryostat, autostainer 
(DAKO  

 
  and Sakura) experience
 
·    Skills with frozen and paraffin sectioning, HE stains and IHC stains
 
·    HT preferred
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Re: [Histonet] unstained paraffin tissue slides storage--why cold?

[dusko trajkovic]

I did an experiment about 10 or 12 years ago, where I cut the sections and 
stained them at various intervals: Day1, Day 15, 1 month, 3 months, 6 months, 1 
year.
Before staining:
Set of slides were allowed to sit at room temp (RT).
Set of slides were stored at 4C.
Set of slides were vacuum sealed and stored at RT.
Set of slides were vacuum sealed and stored at 4C.

Slides stored at 4C consistently stained well, with a slight variance in 
staining after 1 year. Did not matter weather they were vacuum sealed or not.
Slides left out at RT did not fare so well. Staining variability was noticed 
after 1 month. When the slides were stained after one year, signal was almost 
eliminated. Variability and loss of antigenicity was observed with the vacuum 
sealed slides as well.
I did this experiment for just one antibody which was being extensively used at 
the time for one of our projects. It could be that other antibodies fare mach 
better under RT conditions, but why take a chance?

We keep all of our control slides at 4C.
Blocks are kept at RT.

Thanks
Dusko Trajkovic





From: Morken, Tim timothy.mor...@ucsfmedctr.org
To: Helen Fedor hfe...@jhmi.edu
Cc: histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu
Sent: Thu, November 4, 2010 9:09:16 AM
Subject: RE: [Histonet] unstained paraffin tissue slides storage--why cold?

Helen, did you write a paper from that study? 

Tim Morken
Supervisor, Histology, IPOX
UCSF Medical Center
San Francisco, CA, USA

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Helen Fedor
Sent: Thursday, November 04, 2010 8:56 AM
To: sgoe...@xbiotech.com; Emily Sours
Cc: histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] unstained paraffin tissue slides storage--why cold?

The study that we did showed that the staining on freshly cut slides from a 
block stored at room temperature was in fact not as good as slides that were 
sectioned 5 years earlier and stored at -20. Therefore the blocks should be 
stored in the cold as well. The tissue is degrading in the block and on the 
slides and the cold does slow down the process. The fixation in lots of the 
tissue is not optimal. Fixing for 48 hours will definitely  be better than the 
current fixation practices going on in most clinical labs,  if what you want is 
the best tissue preservation possible. But most of us do not have that option.

Helen L. Fedor 

Tissue Microarray Lab, Manager
Prostate Spore Lab, Manager
Johns Hopkins University
600 N. Wolfe St, | Marburg Room 406
Baltimore, MD | 21287-7065

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of 
sgoe...@xbiotech.com
Sent: Thursday, November 04, 2010 11:21 AM
To: Emily Sours
Cc: histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] unstained paraffin tissue slides storage--why cold?

-70 or -80 seems a little extreme to me, that's why I always just leave
them in a normal freezer (-20).  I think the main point of doing this
from what I understand is so that the antigens stay viable.  I know
over time they can degrade and so your stain won't work with some
antibodies.  The weirdest part to me has always been that you don't have
to store the blocks this way.  So I think that was your question, if the
blocks aren't stored in a freezer why store the slides?  Won't the
antigens in the blocks start to degrade as well?  This is a question I
would like to know the answer to as well...

Sarah Goebel, B.A., HT (ASCP)
Histotechnician


XBiotech USA Inc.

8201 East Riverside Dr. Bldg 4 Suite 100

Austin, Texas  78744

(512)386-2907




 Original Message 
Subject: Re: [Histonet] unstained paraffin tissue slides storage--why
cold?
From: Emily Sours talulahg...@gmail.com
Date: Thu, November 04, 2010 7:07 am
To: histonet@lists.utsouthwestern.edu

Can I ask what the point of storing paraffin sections in freezing cold
storage?
They are wax sections, which never see any type of cold, so I don't
understand the point of this. I do understand putting them at 4 degrees
to
prevent mold, but -80 seems excessive.
We have kept our slides at room temperature for years and years, but
these
slides do not have an albumin coat (which I can see getting moldy), just
a
chemical coating.
Fixing for paraffin and paraffin infiltration seems to keep antigens
safe
without refrigeration because it's so intense, but that's just
conjecture on
my part.

Emily
--
Outside of a dog, a book is man's best friend. Inside of a dog it's too
dark
to read.
--Groucho Marx
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[Histonet] CD34 marker for Rat tisue

[dusko trajkovic]

Does anyone have a protocol or any information on CD34 antibody that works on 
FFPE rat tissue?
 
Thank you
Dusko Trajkovic
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Re: [Histonet] DRGs

[dusko trajkovic]

I also think that it is strange of the way Histogel processes. I have posted on 
the Histonet previously about this exact problem. I worked with Jennifer 
Hofecker when she was at Vanderbilt U.(sent her my Histogel and she sent me 
hers) and ended up with perfectly processed Histogel blocks at our facility and 
hers. I processed a couple of blocks last week and they were just terrible. No 
change in the processing schedule, or the way Histogel was liquefied (placed in 
hot water that was heated in the microwave). Prior to the last two blocks, I 
must have processed at least a dozen blocks without any problems. There was an 
incident where I placed two histogels in the same cassete. One processed 
beautifuly and the other was all shrunken and dried up.
I do not liquefy the entire tube, rather I scoop out the approximate amount 
that I need and transfer to another tube to heat up. If there is anyone out 
there in Histoland that has not had any issues with the Histogel, can you 
please post your procedure on liquefying the Histogel, method of 
cooling/solidifying and processing schedule? The only thing that I do that is 
not exact is I do not know the temp of my hot water when i place the Histogel 
to liquefy. I basically have to wait several minutes for the gel to melt and I 
use it immediately.
Any new information or solution from anyone, would be greatly appreciated.
Thank you
Dusko Trajkovic




From: Amy Porter port...@msu.edu
To: Andrea Grantham algra...@email.arizona.edu; HISTONET 
histonet@lists.utsouthwestern.edu
Sent: Mon, February 22, 2010 9:01:22 AM
Subject: RE: [Histonet] DRGs

I think it is strange that we are all doing similar techniques and wind up
with different outcomes using the histogel.  I would be curious how many of
us are using the equipment sold with the histogel for warming and cooling
opposed to any of us who don't.  we did not purchase the equipment and I
wonder sometimes if warming the histogel using other means causes some type
of breakdown / and do any of you repeatedly reheat the same tube once it has
been warmed and resolidified??

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Andrea
Grantham
Sent: Monday, February 22, 2010 9:41 AM
To: HISTONET
Subject: Re: [Histonet] DRGs
Importance: High

Hi Carol,
I have used histogel for these kinds of samples and also other small,  
thin tissues like insect antennae and insect GI tracts and midguts.  
Since I get all my projects already fixed in whatever fixative the  
investigator chooses, rinsed and placed in 70% ETOH the histogel never  
touches formalin. I don't use formalin on my processor but start in  
70%. I've never had a problem with the histogel. We just put the  
sample in the histogel flat and stand it up (turn 90°) when embedding  
in paraffin. I use tissue prep for embedding.
If you don't want to use histogel you could try to put the drg's on GN  
Metricel membrane disc filters. We do this with a lot of the samples I  
receive, actually I have the investigators or their techs do this. The  
tissue sticks to the membrane and orientation is a dream. The membrane  
presents no problem when sectioning. You can get it from VWR.

Andi



Andrea Grantham, HT (ASCP)
Senior Research Specialist
University of Arizona
Cell Biology and Anatomy
Histology Service Laboratory
P.O.Box 245044
Tucson, AZ 85724

algra...@email.arizona.edu
Tel: 520.626.4415    Fax: 520.626.2097

happy slicing and dicing and may all your stains work perfectly -  
Paula Sicurello
P Please consider the environment before printing this email.




On Feb 18, 2010, at 11:23 AM, Barone, Carol wrote:

 Histonetter's...we received a boat-load of mouse DRGs that had been
 prepared in histogel and are cutting...well..not so good.
 We normally do DRGs from FS and get beautiful results.

 We have used histogel before with other small sample and have never  
 had
 issues...  not sure if it is the Histogel or DRG's (fixed in 10% NBF  
 and
 then transferred to 70% before placed into the histogel).is the  
 issue..I
 seem to remember that histogel requires formalin and wonder if the
 transfer to 70% is causing our problem ...but, obviously there is not
 much room for error with such tiny- tiny samples and they are already
 process and in paraffin?

 I am not quite sure how twe can improve the ones that came in  
 histogel,
 and were processed to paraffin a paraffin blockany idea's? any
 experience? any anything? Thx- ASAP!

 cbar...@nemours.org
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Re: [Histonet] Is anyone familiar with Granzyme B antibody?

[dusko trajkovic]

Unfortunately I am trying to get out of the lab and go home to pack for the 
NSH, so I do not have the catalog number, but eBioscinece has a good biotin 
anti mouse Granzyme B cat#13-8822 that works well on mouse tissue. They also 
have a biotin anti mouse F4/80.
Good luck
Dusko





From: Jennifer Campbell jcampb...@vdxpathology.com
To: histonet@lists.utsouthwestern.edu
Sent: Thursday, October 1, 2009 9:35:24 AM
Subject: [Histonet] Is anyone familiar with Granzyme B antibody?

  I'm trying to find out some information on Granzyme B antibody for
immunostaining of FFPE mouse tissue.  Does anyone have any vendors they
could recommend and/or protocols?  

Thanks,

Jennifer
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Re: [Histonet] Best IHC stainer

[dusko trajkovic]

Having worked with Dako autostainer for years and currently using Vantana 
Discovery XT and Leica Bond stainer's, my preference would be the Bond. You 
have the capability of the Ventana stainer, and being able to unlock the 
software that will allow you to use it as any other stainer on the market 
today. Bond also has a favorable per slide cost analysis. If you need any 
additional info, I would be happy to provide as much as I am can.

Thanks
Dusko Trajkovic
Pfizer Inc La Jolla
858-638-6202




From: godsgal...@aol.com godsgal...@aol.com
To: gareth.da...@hotmail.com; histonet@lists.utsouthwestern.edu
Sent: Thursday, August 27, 2009 8:05:35 AM
Subject: Re: [Histonet] Best IHC stainer


Ventana reagents eem to be a little pricey, but if you want a system that you 
can load and walk away from, it is the way to go.  DAKO is an easy enough to 
use instrument and is an open system, but they are going to a price per slide 
kind of thing, if I remmeber correctly.



I prefer the IP from Biocare, as is is an open system and a continuous load 
instrument.  And it mixes the chromagen online


-Original Message-
From: Gareth Blaeuer Davis gareth.da...@hotmail.com
To: histonet@lists.utsouthwestern.edu
Sent: Wed, Aug 26, 2009 8:04 pm
Subject: [Histonet] Best IHC stainer





he lab I work in has been doing demos on IHC stainers Ventana Benchmark XT and 
ako's Autostainer Plus.  We could really use feed back
from current users of both.  We are having a hard time deciding between the 
two, 
o any input would be great.
What are the Pros and Cons with both.
Thanks,
Gareth Blaeuer Davis, B.S., HT

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Re: [Histonet] CD31 working protocol

[dusko trajkovic]

Hi Patricia,
I use an antibody from Spring Biosciences Cat#M3384 to stain our human 
xenograft tumors as well as human tissue. Antibody will also stain mouse 
tissue. Very nice clean crisp stain. At least when we use it on our Bond IHC 
stainiers.
Thanks and let me know if you need more information.
dusko





From: Zerfas, Patricia (NIH/OD/ORS) [E] zerf...@ors.od.nih.gov
To: dusko trajkovic dunat...@sbcglobal.net
Sent: Monday, August 10, 2009 12:07:29 PM
Subject: RE: [Histonet] CD31 working protocol


Dusko,
    I am using mouse tissue.
 
Patricia Zerfas
National Institutes of Health
Building 28A, Room 112
28 Library Drive
Bethesda, MD   20892
ph:   (301) 496-4464
fax:  (301) 402-1068
 



From:dusko trajkovic [mailto:dunat...@sbcglobal.net] 
Sent: Monday, August 10, 2009 3:06 PM
To: Zerfas, Patricia (NIH/OD/ORS) [E]
Subject: Re: [Histonet] CD31 working protocol
 
What species are you trying to stain?
thanks
Dusko
 



From:Zerfas, Patricia (NIH/OD/ORS) [E] zerf...@ors.od.nih.gov
To:  histonet@lists.utsouthwestern.edu   histonet@lists.utsouthwestern.edu 
Sent: Monday, August 10, 2009 11:50:50 AM
Subject: [Histonet] CD31 working protocol

Dear Listservers,
            I have tried MANY protocols and I am unable to obtain a working 
protocol for the CD31 antibody.
            I have tried both the CD31 rat monoclonal and a CD31 rabbit 
polyclonal.

Thanks in advance for your help.

Patricia Zerfas
National Institutes of Health
Building 28A, Room 112
28 Library Drive
Bethesda , MD   20892
ph:  (301) 496-4464
fax:  (301) 402-1068

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[Histonet] Can I get some help with Survivin ab.

[dusko trajkovic]

Hallo Everyone,
For those of you who have used this antibody on FFPE human breast tissue, can 
you provide me with a protocol and antibody information? I have tried 
antibodies from Cell Signaling, Spring and two from Biocare, however I am not 
getting the results that are the same as in the publications.
Any help/response would be greatly appreciated.
thanks you and have a very good day.
 
Dusko Trajkovic
DSRD Scientist 
Pfizer Inc La Jolla
858-638-6202
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Re: [Histonet] Ventana Reagent Cost??

[dusko trajkovic]

Ventana prices will vary dependednt upon your (not you per se, but rather your 
instrument) consumption.
Here is what we pay:
LCS    $74
CC1    $832
CC2    $895
EZprep (10X) $452
Reaction Buffer$65
If you need more info, please contact me

Thanks
Dusko Trajkovic
Pfizer Inc La Jolla





From: Burton, Lynn lynn.bur...@illinois.gov
To: Tiana Fountain tfount...@exchange.hsc.mb.ca; 
histonet@lists.utsouthwestern.edu
Sent: Friday, June 19, 2009 9:46:37 AM
Subject: RE: [Histonet] Ventana Reagent Cost??


The prices I have are on the list you sent. I couldn't find the CC2 price 
quickly.
Lynn Burton
Lab Assoc. I
Animal Disease Lab
Galesburg, Il 61401



From: histonet-boun...@lists.utsouthwestern.edu on behalf of Tiana Fountain
Sent: Fri 6/19/2009 10:42 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Ventana Reagent Cost??



Hello everyone,

I am looking for a cost of some of the Ventana reagents can anyone help?
Specifically:

LCS:$98.40
CC1:$565.80
CC2:
EZ Prep:$37.80
Reaction  Buffer:$43.80

I am not currently a customer so I didn't want to phone Ventana directly
if I don't have to. Thanks

Tiana



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