Re: [Histonet] lot to lot

2011-08-29 Thread Patti Loykasek
I know different people do lot to lot checks in various ways. We use 
concentrated antibodies, and so we like to do a 'sandwich' titer check on the 
new lot using our usual tissue control tissue. For example, if the last CD20 
lot was at 1:100 we would check the new lot at 1:50, 1:100, 1:200 using tonsil 
tissues  our standard SOPs. I did have 1 inspector tell me they thought that 
was unnecessary. It's our preference to do 3 titers  assess the maximum 
performance in one fell swoop. Our detection lot to lot check is completely 
different. We check 8-10 different antibodies using our standard control tissue 
with the new detection lot. we use antibodies with a variety of cellular 
expressions to assess cytoplasmic, membranous  nuclear expression. We make 
sure to include some of our 'fussy' antibodies. Of course we have SOPs  forms 
to go along with this. 
Call me crazy - I prefer to think of it as meticulous  dedicated! We have had 
a couple of problems with detections over the years. 
Just my 2 cents. 

Patti Ann Loykasek HTL, QIHC
PhenoPath Laboratories


On Aug 29, 2011, at 9:06 AM, Mary Helie wrote:

 Good Afternoon
 
 I would like to get an idea of how many slides people are running for lot to 
 lot antibody and detection testing. For example would a tonsil with good 
 internal negative and positive elements be good enough to validate a new lot 
 of LCA?
 
 Thank you
 Mary
 
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[Histonet] position opening

2011-08-09 Thread Patti Loykasek
Due to an upcoming retirement, PhenoPath has a position opening in the contract 
research department. This position is as described below. I'm happy to answer 
any questions on this position. 


Contract Research Laboratory Supervisor
PhenoPath Laboratories, PLLC, has an opportunity for a Laboratory Supervisor in 
its Contract Research Department. This is a supervisory position located in 
Seattle, WA and reports to the Director of Contract Research.
Primary Responsibilities
Responsibilities include, and are not limited to: Project Development – 
managing all aspects of client requests for RD projects to include project 
proposals, reporting project status, and resolution of questions; Project 
Execution – oversee and coordinate work of Research Technologists to include 
assay development, specimen tracking and testing, and quality review of slides; 
Supervision of Contract Research personnel – staff training and development, 
performance management, assist in recruiting efforts for new staff.
Required Skills/Experience
Requires 8-10 years of Immunohistochemistry experience and 3-5 years of 
supervisory and project management experience; preference for an HTL or an MT 
with experience working in a clinical laboratory or GLP laboratory setting.
TO APPLY, PLEASE SEND COVER LETTER AND RESUME TO:
PhenoPath Laboratories, PLLC 551 N. 34th St., Suite 100 Seattle, WA 98103 
Phone: 206 374-9000
Fax: 206 374-9009 E-mail: j...@phenopath.com



Patti Loykasek
PhenoPath Laboratories
Clinical Lab Manager
Seattle, WA

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Re: [Histonet] IHC Patient Negatives

2009-10-28 Thread Patti Loykasek
Well, I hope I don¹t start a controversy, but here's my 2 cents worth. I
don't think you should run another negative. If you are going to have any
staining due to intrinsic tissue elements reacting with your detection, you
will see it the first day. I am of the opinion that in most cases too many
negative controls are run. You are wasting precious patient material. With
today's polymer detection systems, there is very little background staining.
If you are doing additional testing on a patient using a different type of
detection, I do think you should run another negative.

Please don't flame me too badly - I'm having a bad day! And it is only my 2
cents worth!


Patti Loykasek BS, HTL, QIHC
Clinical Lab Supervisor
PhenoPath Laboratories
Seattle, WA






 Hi All,
 
 
 
 I have an issue with my computer, so I cannot see past the little
 paragraphs on the Histonet archives.  Sorry!  This question is probably
 asked many, many times.
 
 
 
 When we run our IHC's we always have a patient negative slide to go with
 our case.  We run everything accept Herceptin on the Bond. When running
 a patient's IHC on the bench and on the Bond we use a negative patient
 control for both.  (We run a test control on all antibodies also)  If we
 are running a double stain IHC also we run another negative patient
 control for that stain also.  The debate we are having is, if the next
 day you run another IHC on the same patient using the same DAB define
 kit, should we be running another negative patient control?
 
 
 
 Thanks for your help!
 
 
 
 Josie Britton HT
 
 Cheshire Medical Center
 
 
 
 
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[Histonet] Plants in lab

2009-10-22 Thread Patti Loykasek
Hi All.  Just to clarify - the inspection we had was not a CAP inspection.
It was an audit based on the GLP regulations.
Thanks for all the feedback on plants in the lab.

Patti Ann Loykasek
PhenoPath Laboratories



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[Histonet] Inspection question

2009-10-21 Thread Patti Loykasek
Hi All. Happy Wednesday. Has anyone everyone had an auditor/inspector note
that plants in the histology laboratory are a possible contamination hazard
 must be removed? Just wondering.


Patti Loykasek BS, HTL, QIHC
Clinical Lab Supervisor
PhenoPath Laboratories
Seattle, WA




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Re: [Histonet] HerCep

2009-10-14 Thread Patti Loykasek
Depending on the tissue fixation  processing, you can see some cytoplasmic
staining. How does the cell pellet control look? We seldom see cytoplasmic
staining on that. Are you tracking that your retrieval buffer is 95-100
after you add the slides  starting your retrieval times when the buffer
temp gets back up to 95? Wash well between all incubations, too.
That's just some thoughts off the top of my head. Let me know if you have
more specific questions on the protocol.


Patti Loykasek BS, HTL, QIHC
Clinical Lab Supervisor
PhenoPath Laboratories
Seattle, WA




 Does anyone have any experience using the Dako HerCep kit?
 
 And do you have any problems with cytoplasmic staining?
 
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Re: [Histonet] looking for calibration tools for Optimax plus stainer

2009-10-13 Thread Patti Loykasek
In reference to LeRoy's last statement below- if anyone is selling time
please put me on the list for the maximum allowable! I haven't been able to
figure out how to cram more hours in a day!
And it's only Tuesday.

Patti Loykasek



 Does anyone have a set of calibration tools  needed to calibrate my BioGenex
 Optimax stainer.  Seems they no longer  make these or sell time??
 
 thanks  
 
 LeRoy Brown Ht(ASCP) HTL
 
 www.histocs.com  
 
 360-966-7300
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[Histonet] Oscar

2009-09-28 Thread Patti Loykasek
Hi All. I understand that there was a recent question regarding the antibody
Oscar (pan cytokeratin). I've been off histonet while I was on vacation. We
routinely work up Oscar using normal liver. There should be staining of the
bile ducts and the hepatocytes. I'd be happy to offer more info if the
person who originally posted will contact me. I'll try not to take vacation
again ( at least until the next available time!). Happy Monday.

Patti Loykasek
PhenoPath Laboratories



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[Histonet] Position open in Seattle

2009-07-30 Thread Patti Loykasek
Due to recent growth and new projects, we have a position open in Seattle.
Here is the announcement:
HISTOTECHNOLOGIST
 
PhenoPath Laboratories, PLLC, a pathology reference laboratory, has an
opening for a Histotechnologist (full-time, M-F.) We are in a
state-of-the-art facility located on the scenic Ship Canal in Seattle,
Washington.
 
Primary Responsibilities
Responsibilities include performing routine manual and automated tests in a
clinical and research histology laboratory. Under general supervision,
perform routine laboratory tests such as processing, embedding, microtomy,
frozen sections, HE, special stains, routine IHC, etc.
 
Required Skills/Experience
Requires ASCP certification (or certification-eligible) Histotechnologists.
PhenoPath Laboratories is committed to hiring the best person for the job.

PhenoPath Laboratories is a national specialty pathology laboratory
committed to providing outstanding clinical and research services. We offer
an excellent compensation and benefits package, including 401k.  Salary will
be determined based on the overall skills and background of the chosen
applicant. 
 
To Apply Please Contact:
PhenoPath Laboratories, PLLC
551 N. 34th St., Suite 100
Seattle, WA 98103 
Phone: 206 374-9000
Fax: 206 374-9009 
E-mail:j...@phenopath.com
 
Please no phone calls about this position from candidates or recruiters.


Patti Loykasek BS, HTL, QIHC
Clinical Lab Supervisor
PhenoPath Laboratories
Seattle, WA




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[Histonet] Beecher tissue array maker

2009-07-27 Thread Patti Loykasek
Hi All. I have been attempting to contact Beecher Instruments regarding an
ETA on TMA needles. So far, no replies to voice mail or email. Does anyone
have a personal contact at this company? Thanks.


Patti Loykasek BS, HTL, QIHC
Clinical Lab Supervisor
PhenoPath Laboratories
Seattle, WA




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[Histonet] Slide dryers

2009-07-22 Thread Patti Loykasek
Hi All. We are re-working some of our processes (fun!). We are trying to get
more LEAN and shave some time off of our processes here  there. One thing I
am looking at is the small forced air slide dryers. I have seen some good
comments on the mopec roto-dry. Has anyone tried the dryer from EMS? What
are people using and liking? We will be doing IHC on these sections. What
type of drying time are you using? I really appreciate the info. We are
having a series of quick meetings, and I am having to gather information
quickly. Thanks for the help.



Patti Loykasek BS, HTL, QIHC
Clinical Lab Supervisor
PhenoPath Laboratories
Seattle, WA




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[Histonet] HTL/BS

2009-07-14 Thread Patti Loykasek
Thought I would throw my 2 cents out there, too. I received my HTL back in
the early 80's (hate to think how old that makes me!), I already had a BS
when I went to histotech school. I have found that it has made a difference
in my career to have the HTL. When I have worked in hospitals, I was on the
same pay scale as the med techs, the HT's were not. When I received my QIHC,
some kind med techs lobbied for me to get the same specialty certification
bonus that the med techs received. I feel fortunate to have mostly been
treated by physicians and co-workers with respect, and do feel that my work
is a profession ( I enjoy it). I am thankful  to have the wonderful
experiences, and I go home  have a glass of wine on the bad days!

Patti Loykasek



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[Histonet] Bone marrows

2009-07-10 Thread Patti Loykasek
Hi All. I was wondering what everyone's current favorite bone marrow decal
solution/method is. Including commercially available decal solutions. We
will be doing IHC after fixation/decal. Thanks for the input (as always).


Patti Loykasek BS, HTL, QIHC
Clinical Lab Supervisor
PhenoPath Laboratories
Seattle, WA




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[Histonet] Job opening

2009-07-08 Thread Patti Loykasek
HI All. We currently have a job opening for a technologist in our molecular
testing section. Here is the job posting (it is on our web site, too):

Molecular Technologist (full time):
PhenoPath Laboratories, PLLC, a national pathology reference laboratory, has
an opportunity for a Medical Technologist or Histotechnologist in our
Molecular section (full-time, M-F).

Primary Responsibilities:
Responsibilities include: Performing FISH and/or PCR on paraffin specimens,
blood/bone marrow and other clinical specimens. Maintaining QA/QC data and
writing/reviewing SOPs.

Required Skills/Experience:
Requirements include: ASCP certification in Medical Technology or
Histotechnology
required. Prefer two-plus years of molecular experience in a clinical
setting; strong
interpersonal skills.

TO APPLY, PLEASE SUBMIT A COMPLETED APPLICATION FOR EMPLOYMENT
AND RESUME TO:
PhenoPath Laboratories, PLLC
551 N. 34th St., Suite 100
Seattle, WA 98103
Fax: 206 374-9009
E-mail: j...@phenopath.com
Application for employment may be downloaded from www.phenopath.com
Please no phone calls regarding this position.


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[Histonet] Used equipment dealers

2009-06-29 Thread Patti Loykasek
Hi all. I would appreciate any info on used histology equipment dealers (
such as tissue processors) on the West Coast. Thanks for the help.


Patti Loykasek BS, HTL, QIHC
Clinical Lab Supervisor
PhenoPath Laboratories
Seattle, WA




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Re: [Histonet] DAB substrate buffer

2009-06-26 Thread Patti Loykasek
Hi Dr. Kumar. We use a 0.05M Tris buffer pH 7.7-7.9. We add 3% H2O2 right
before use. We use our own stock DAB. Perhaps your DAB was too dilute? I
hope this info helps.  If you would like more specifics, you can contact me.


Patti Loykasek BS, HTL, QIHC
Clinical Lab Supervisor
PhenoPath Laboratories
Seattle, WA

 
 hi every one,
   I am using Labvision HRP polymer secondary antibody with DAB
 chromogen, funny thing is that i have exhausted the DAB substrate buffer. so i
 tried with PBS 7.4 and h2o2 and chromogen and i found out that nothing worked.
 can anyone tell me a protocol or recipe for preparing DAB substrate buffer.
 
 Regards,
 Dr. Anjan Kumar.K.R
 
 M.V.Sc Scholar
 
 Dept. of Veterinary Pathology
 
 Madras Veterinary College
 
 Chennai-7
 
 India
 
 email: drvet_an...@hotmail.com
 
 Phone: +91-9940475801
 
 
 
 _
 Live Search extreme As India feels the heat of poll season, get all the info
 you need on the MSN News Aggregator
 http://news.in.msn.com/National/indiaelections2009/aggregator/default.aspx
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Re: [Histonet] IHC on fresh frozen

2008-12-03 Thread Patti Loykasek
After hearing a presentation by Sharon Lear describing some low temp antigen
retrieval that she did, we changed our method for frozen sections. We fix
the frozen sections in 10% nuetral buffered formalin for 30'-60', rinse,
then do a gentle pretreatment. The gentle pretreatment is usually 10mM
citrate buffer pH6 at 70 degrees C for 30'. Slides are cooled, and usual IHC
done. This has worked well for us. Our staining with this method is more
reliable  intense than with previous methods.

Patti Loykasek



 Hi, all
 
 Recently I did some IHC(chromagen methods) on mouse fresh frozen
 tissues, mainly using insulin antibody on pancreas. The image is
 much fuzzier compare to paraffin embedding tissue. And the
 staining also smeared to acinar cells which surround the islet.
 
 I airdried slide(30min) and used a general acetone method(-20C
 5min) to fix the tissue before I did IHC.
 
 How can I get a relatively sharp staining on the fresh frozen
 tissue?Does anyone here have any experience on it? Any
 suggestions?
 
 Many thanks and have a nice day,
 
 Ann
 
 
 
 
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[Histonet] Thanks new labeler question

2008-12-03 Thread Patti Loykasek
Thanks to all for the answers on my expired reagents. Now, I have a new
question. Is anyone using the Brady label printer or another similar printer
for slide labels? I would be interested in hearing your experiences. Thanks,
as always. 

Patti Loykasek



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[Histonet] ERCC1 antibody

2008-11-12 Thread Patti Loykasek
Hi all. Just wondering if someone would share what antibody they are using
for ERCC1. We have used clone 8f1 in the past, but was wondering if perhaps
there is a new, better clone. I haven't been impressed with the staining
that we have. Thanks.


Patti Loykasek BS, HTL, QIHC
Clinical IHC Supervisor
PhenoPath Laboratories
Seattle, WA




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Re: [Histonet] experiences with kappa/lambda

2008-11-12 Thread Patti Loykasek
Hi Gudrun. We have a fair bit of success with kappa/lambda IHC. We use
polyclonal antibodies, a heat citrate pretreatment. The caveat is that we
have several different titers in use - we have a titer for lymph nodes, one
for bone marrows/decaled tissue, one for tissue fixed in zinc formalin,
etc We have pretty good correlation with our flow lab. I believe that a
couple of years ago Rich Cartun published a paper on his methods  he has
good correlation, too. It does require a pathologist with good knowledge of
heme path  patience to work out the optimum staining with the lab.


Patti Loykasek BS, HTL, QIHC
Clinical IHC Supervisor
PhenoPath Laboratories
Seattle, WA




 Hi listmembers,
 
 Please tell me about your experiences with the IHC-demonstration of kappa
 and lambda on FFPE-sections.
 
 We have some troubles about inconsistend staining and therefore false
 results about the monoclonality of tumors.
 
 Bye
 
 Gudrun
 
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Re: [Histonet] Histology or IHC conferences/workshops

2008-10-01 Thread Patti Loykasek
Hi Igor. I am not sure of specific topics for 2009 but you should look at
the websites of ASCP, NSH, and TNT (teleconference network of TX). All of
these entities sponsor teleconferences. NSH will be in Birmingham, AL 2009 
there are always great seminars there.

Patti Loykasek


 Dear Histonetters!
 I was wondering where someone knows of any useful conferences or workshops ,
 which deal with histological techniques, or Immunohistochemical techniques
 or even microscopy techniques( I heard about a place in Cape Cod), coming up
 in 2009??? Any information would be appreciated.
 Thank you.
 Igor Deyneko
 Infinity Pharmaceuticals
 In-Vivo Pharmacology
 Cambridge, MA 02139
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