[Histonet] Immuno. confusion

[Amos Brooks]

Hi,
Well... almost any point before DAB. You wouldn't want to put it in
after the HRP conjugated secondary because that would quench the HRP signal
you are actually looking for leaving you nothing for the DAB to precipitate
on. On the other hand you certainly do not *need* to do it after the HIER as
it will work just fine before it or even after the primary antibody
(assuming it isn't HRP conjugated). Actually we have had the best results on
CD4 if the H2O2 is done prior to the HIER. The H2O2 in this case actually
affects the epitope if you do it after HIER. If it does that with CD4, I
assume it is likely to do so with other finicky epitopes.

Have a nice day,
Amos

On Wed, Aug 3, 2011 at 1:00 PM,
wrote:

> Message: 7
> Date: Wed, 3 Aug 2011 11:49:40 -0500
> From: "Troutman, Kenneth A" 
> Subject: [Histonet] Immuno. confusion
> To: "Histonet@lists.utsouthwestern.edu"
>
> Message-ID:
><
> 7b310892042da74cb3590053f424cfe6143ee30...@its-hcwnem06.ds.vanderbilt.edu>
>
> Content-Type: text/plain; charset="us-ascii"
>
> Hi Valerie,
>
> The peroxidase blocking step is to prevent endogenous peroxidase activity
> that will cause a false positive stemming from your application of DAB (the
> substrate for your DAB solution is hydrogen peroxide).  You can essentially
> put this blocking step at any point in your stain (following
> deparaffinization) and before your application of DAB.  I have run this step
> at varying points in my procedures and they all work fine.  Some antibodies
> prefer you do this step before primary application and others prior to DAB,
> but for most stains, it doesn't matter too much.
>
> As for an enhancer, you can make a copper sulfate solution (5mg/mL in
> buffer).  Or you can try DAB Enhancer or DAB Sparkle from Biocare.
>
> Good luck,
>
> Ashley Troutman BS, HT(ASCP) QIHC
> Immunohistochemistry Supervisor
> Vanderbilt University Histopathology
> 1301 Medical Center Drive TVC 4531
> Nashville, TN  37232
>
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[Ruppert, Amysue]

We moved into a new lab a few years ago, and due to the incoming air current 
with the ventilation here, it caused our cyrostats to ice up considerably. We 
have the type of ventilation that has air vents on the ceiling and are 
constantly blowing large amounts of air into our rooms. We had to get a piece 
of plastic put into the vent to divert some of the air away from our cryostat. 
It helped, but still ices more than when we were in our old lab. Anyway, point 
is to check out the air flow above the cyrostat. You also need to have enough 
clearance behind the cryostat and on each side. The clearance amount would be 
listed in the manual. 
good luck
amysue

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[Histonet] microtome service/repair Denver area

[Elizabeth Chlipala]

Hello everyone

I was wondering if anyone has any recommendations for a vendor that does yearly 
service and repair on Leica microtomes and cryostats.  We have been working 
with one vendor for years now and always have been happy with their service but 
they have become very expensive, any suggestions would be appreciated.

Thanks in advance

Liz

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Manager
Premier Laboratory, LLC
PO Box 18592
Boulder, CO 80308-1592
(303) 682-3949 office
(303) 682-9060 fax
(303) 881-0763 cell
www.premierlab.com

Ship to address:

1567 Skyway Drive, Unit E
Longmont, CO 80504

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[Ruppert, Amysue]

One point to bring up, and I am sure that Leica service will do this, is the 
brand of knives you are using. Different brands have slightly different 
thickness. This is not related to the height of the blade. We have found that 
we need to use the blades that are used by our service company to calibrate the 
microtomes to have the correct tension in the holder and not any problems with 
the quality of the sections. 
 good luck, seems you are on the right track though.
amysue ruppert

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Re: [Histonet] Manual embedding

[Stella Mireles]

A couple of ideas:
Suggest looking for a used paraffin bath from an outside vendor. #2 Idea: If
anyone knows of a lab that is closing up, they might be able to contact
Andrea and pass that info. I know when a lab is closing up, storage of
equipment becomes burdensome.
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Re: [Histonet] Manual embedding

[Rene J Buesa]

Stella:
Just remember that the mission of HistoNet is not only to reply to specific 
queries, but to do so in a way that we all could benefit.
Instead of sending a good idea to somebody privately, you should let everybody 
know about it.
René J.

--- On Wed, 8/3/11, Stella Mireles  wrote:


From: Stella Mireles 
Subject: Re: [Histonet] Manual embedding
To: "Andrea Marion" 
Cc: histonet@lists.utsouthwestern.edu
Date: Wednesday, August 3, 2011, 4:29 PM


Andrea,
I have send you an idea that might work for you. Check your email.
Hope this will help.
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Re: [Histonet] Manual embedding

[Stella Mireles]

Andrea,
I have send you an idea that might work for you. Check your email.
Hope this will help.
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[Histonet] For Sale - Mopec MB100 Countertop Grossing Station

[American Resource Medical]

For Sale - Mopec MB100 Countertop Grossing Station
 
This is a used piece in excellent condition and half the price of new. 
 
 
 
http://media.mopec.com/media/pdf/Catalog2007smaller159.pdf
 
 
 
This is a countertop unit.   
 
 
American ReSource Medical 
 
P: 201.833.1550

 

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[Histonet] FDA Reagent Label Requirements for Richard allan Flex alcohols

[Wilson, Carol]

Hi All,
I currently got inspected by my QA dept. and they complained that our Flex 
alcohols did not have storage requirements on their labels, is anyone using the 
new Flex alcohols in the larger bottles and if so, do they have the storage 
requirements on the labels.  If not, are there any GLP labs out there that add 
a label to their alcohols when they are received stating storage requirements?  
Looking for ideas to have a simple fix for this.
Thanks,
Carol

Carol Wilson, HT(ASCP)
Lead Technician/Histology
Ricerca Biosciences, LLC

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RE: [Histonet] Immuno. confusion

[pruegg]

   I have seen this done many ways, some people have h202 at th= e end of
   their  depara  setup  if  doing  that  off line.  For the Leica Bon= d
   instrument  we were advised to do h202 block just before the DAB and i
   hav=  e  noticed  that it works best that way for their system.  I for
   years  h=  ave  always  used h202 after the primary ab as some abs are
   affected  by  it so= I just don't take the chance of not knowing which
   may  be.   There  was = one ab, cd4 I believe that for whatever reason
   needed to be h202 blocked be= fore AR and the AB, and it really didn't
   work  if  you  did not do the block b= efore everything.  Just goes to
   show that each ab has to be handled sp= ecifically.

   




   


   A  little  aside  here and techni= cally it makes no sense but we were
   having  trouble  with  the  Leica  ap/red  ki=  t preciptating and was
   advised  by  Jim Burchette not Leica to use H202 = just before the red
   substrate chromogen and that has really improved the ap= /red staining
   for  us  which  we use a lot on derm samples.  I know = endogenous alk
   phos uses levamisol as a block and not h202 but for some rea= son this
   works so I don't argue with it.

   




   


   R= egards,

   


   Patsy

   




   


 


      Original   Message   Subject:   [Histonet]  Immuno.
   confusion
   From: "Hannen, Valerie" >
   Date: Wed, August 03, 2011 8:57 am
   To: "[1]histonet@lists.utsouthwestern.edu"
   &= lt;[2]histo...@lists.utso= uthwestern.edu>
   Hi folks...
   I  am going to try to revam= p out Immuno. procedure and am looking at
   a  couple  of  spec.  sheets  from  our=  primary  vendor. I am gettin
   conflicting information...so I am turning to y= ou all for help.
   When doing immuno's that require either HIER or enz= yme digestion, do
   you  perform  the  retrieval  first or do you do the "peroxid= e" step
   first??
   One   spec.   sheets   says   retrieve   then  block  endogenous  per   
oxidase...the  other says block endogenous peroxidase then do the HIER
   or en= zyme digestion..which is correct??
   One  other question that I have is= ...what do you use to enhance your
   stain??  The  enhancing  solution  that  we  w=  ere  using  has  been
   discontinued.
   Thanks so much!!
   Valerie Han= nen,MLT(ASCP), HTL,SU(FL)
   Histotechnologist
   Parrish Medical Center
   951 N. Washington Avenue
   Titusville, Florida 32796
   (321) 268-6111 ex= t. 7506
   = **
   "This email is intended solely for the use of the individual = to
   whom it is addressed and may contain information that is
   privilege= d, confidential or otherwise exempt from disclosure
   under applicable law= . If the reader of this email is not the
   intended recipient or the emplo= yee or agent responsible for
   delivering the message to the intended reci= pient, you are
   hereby  notified that any dissemination, distribution, or<= BR>copying
   of this communication is strictly prohibited. If you
   have rec= eived this communication in error, please immediately
   delete this messag= e. Thank you"
   **= 
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References

   1. 3D"mailto:his   2. 3D"mailto:histonet@lists.utsouthwestern.edu";
   3. 3D"mailto:Histonet@lists.utsouthwestern.edu";
   4. 
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Re: [Histonet] Manual embedding

[Andrea Marion]

Hi Scott,

I work in a research lab that does manual embedding too. I looked into
purchasing a paraffin pitcher, but it was too expensive for us. Our
makeshift embedding center is comprised of:

1) 3 thick-walled plastic 500 ml beakers covered in a double layer of foil
(you could use glass, but it gets slippery with melted paraffin on it)
2) a heating block with the surface covered in aluminum foil
3) a water bath
4) a set of weighted rings

We remove most of the water from the bath (otherwise the paraffin beakers
want to float), and use the beakers to store melted paraffin. Two are for
infiltrating, one stores clean paraffin for embedding. The weighted rings
are placed on top of the beakers, supported by the foil. The foil also
keeps water from getting into the paraffin.

After infiltration, I use a plastic mold, place it on the covered heating
block (held at ~65 degree C), fill with paraffin, then place and orient
the specimen in the mold. The blocks sit at room temperature until they
are mostly set, then stored in a 4 degree fridge to finish hardening.

The only problem we have had is occasionally getting water in the paraffin
from condensation. This hasn't been a problem since removing the lid of
the water bath. If it happens, you will see the water sitting under the
melted paraffin. We either replace the paraffin, place plastic cassettes
at the bottom of the beaker so the samples can sit above the level of the
water, or use disposable plastic pipettes to remove the water.

I have also used a dry incubator set to 60 degrees with the same
equipment. It is slightly more convenient, but the paraffin takes longer
the melt than in the water bath (heat exchange is slower).

I also occasionally use a metal pitcher for storing paraffin either in the
incubator or water bath (heating jacket has not been necessary for us).
You can purchase these very cheaply from Amazon - look for stainless steel
milk frothing pitchers that are used for making coffee drinks.

Good luck!

Andrea Marion
Graduate Student
University of Illinois at Chicago


Dear Histonetters:

I am interested in acquiring a pitcher and heating jacket for melting and
pouring paraffin during manual embedding. My work is relatively low volume
and in a university research lab setting so I am trying to avoid purchasing
an expensive embedding station. Can anyone recommend an honest supplier of
used histology equipment that might be able to provide me with this item?

Thank you for your expertise!

Scott L. Parker


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[Histonet] Immuno. confusion

[Troutman, Kenneth A]

Hi Valerie,

The peroxidase blocking step is to prevent endogenous peroxidase activity that 
will cause a false positive stemming from your application of DAB (the 
substrate for your DAB solution is hydrogen peroxide).  You can essentially put 
this blocking step at any point in your stain (following deparaffinization) and 
before your application of DAB.  I have run this step at varying points in my 
procedures and they all work fine.  Some antibodies prefer you do this step 
before primary application and others prior to DAB, but for most stains, it 
doesn't matter too much.

As for an enhancer, you can make a copper sulfate solution (5mg/mL in buffer).  
Or you can try DAB Enhancer or DAB Sparkle from Biocare.

Good luck,

Ashley Troutman BS, HT(ASCP) QIHC
Immunohistochemistry Supervisor
Vanderbilt University Histopathology
1301 Medical Center Drive TVC 4531
Nashville, TN  37232

From: Hannen, Valerie 
Subject: [Histonet] Immuno. confusion
To: "histonet@lists.utsouthwestern.edu" 
Date: Wednesday, August 3, 2011, 11:57 AM


Hi folks...

I am going to try to revamp out Immuno. procedure and am looking at a couple of 
spec. sheets from our primary vendor. I am gettin conflicting information...so 
I am turning to you all for help.

When doing immuno's that require either HIER or enzyme digestion, do you 
perform the retrieval first or do you do the "peroxide" step first??
One spec. sheets says retrieve then block endogenous peroxidase...the other 
says block endogenous peroxidase then do the HIER or enzyme digestion..which is 
correct??

One other question that I have is...what do you use to enhance your stain?? The 
enhancing solution that we were using has been discontinued.

Thanks so much!!

Valerie Hannen,MLT(ASCP), HTL,SU(FL)
Histotechnologist
Parrish Medical Center
951 N. Washington Avenue
Titusville, Florida 32796
(321) 268-6111 ext. 7506

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[Histonet] job openings

[Tammy de Leon]

 

 

If you're looking for a friendly environment, a new state-of-the-art
facility and an employer of choice, SPL is the place for you!! We are
currently looking for qualified Histotechnicians/Histotechnologists!

 

Histo candidates should be an HTL or HT (ASCP) or equivalent.

Primary responsibilities include on-site frozen sections including mobile
laboratory units.

 

SPL is CAP accredited, offers competitive pay, and a comprehensive benefits
package.  SPL pays 100% of employee premiums for  Medical, Dental, LTD and
Life!  We also offer a Retirement Plan & Supplemental Insurance.

 

Please forward resume to

Tammy de Leon

ta...@surgicalpathlabs.com

Surgical Pathology Laboratory

800-304-1066

8455 66th Street N

Pinellas Park, Florida 33781

www.surgicalpathlabs.com

 

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[Histonet] RELIA Histology Careers Bulletin 8/3/2011 How are you staying cool during these dog days of Summer?

[Pam Barker]

Hi Histonetters!,

I hope you are having a great day!  ! What are you doing to stay cool
during these dog days of summer?
I am sipping icy cold drinks and spending as much time as possible at
the beach and in the Air Conditioning!!!  If you are contemplating
making a job change let me help while you kick back relax and enjoy a
glass of iced tea or lemonade in the shade.  I can keep you posted on
opportunities as they come open, assist you with your resume and coach
you through the interview process.

All of the positions I work with are fulltime 40 hour per week positions
with top hospitals, labs and doctors offices.  My clients offer
excellent compensation including competitive salaries, great benefits,
relocation assistance and in some cases sign on bonuses.  My services
are FREE of charge to you.  All of my fees are paid by my clients, the
facilities that I represent.  I work with companies nationwide that are
in need of histology supervisors, histotechnologists and
histotechnicians.  




Here is a list of my most exciting current openings:



HISTOLOGY/PATHOLOGY  MANAGEMENT

TX – Anatomic Pathology Laboratory Manager - Austin

MA – Night Shift Supervisor – Boston area

NC – Laboratory Manager – Histology/Cytology - Asheville

 

HISTOTECHS

FL – Evening Lead DermpathTech - Miami

NC – Afternoon Shift Grossing Histotechnologist-Charlotte

TX – Histology Technician - Austin

LA – Night Shift Histology Tech - Lafayette

NC – Day shift - Asheville

FL – Mohs Histotech – Sarasota

MD – Night Shift Grossing Histotechnologist - Baltimore

MA – Day shift – Cape Cod 

OTHER OPPORTUNITIES

Lead Grossing Tech – Austin, TX

Path Assistant – Charlotte, NC

Specimen Processing Manager – Portland, ME

If you are interested in any of these positions please call me at
866-607-3542 or e-mail me at rel...@earthlink.net  If you would like you
can e-mail me your resume and a number where I can reach you at a time
that is convenient for you.  If you are interested in looking into new
job opportunities in other areas that are not mentioned above please
contact me as well.  I work with facilities nationwide and I will keep
your resume confidential.  I will only represent you to jobs you tell me
you are interested in looking into.



Remember. It never hurts to keep an eye open even if you are happy in
your present job.


Also if you know anyone else that might be interested I would really
appreciate it if you would pass my information along to them as well.  I
offer a 500.00 referral fee if you refer someone that I place!

Thank - Pam - 866-607-3542 (866-60RELIA)


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

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[Histonet] Dyes used in Crayola wax crayons

[Bob Richmond]

I started what turned into a remarkably lengthy and diverse Facebook
thread about Crayola® brand wax crayons. Some people use them to dye
fabrics, particularly cotton T-shirts.

Poking around the Web, I get the idea that Crayolas are colored by
Procion MX reactive cotton dyes, which come in a vast variety of
colors. The dyes are suspended in particulate form in the wax, rather
than dissolved. Some people fix the dye in the fabric after dyeing by
soaking the fabric in salt water, a practice that certainly sounds
like reactive cotton dyes.

Anyone know if this information is correct? Obviously reactive cotton
dyes don't have much use in histology.

If anybody wants to friend me on Facebook, I'm the Bob Richmond with
"Harvard" after his name.

Bob Richmond
Samurai Pathologist
Knoxville TN

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[Histonet] Seeking Histotechnologist in Upstate NY

[Darcy Bloch]

   Slone=  Partners  is  seeking  experienced  Histotechnologists and new
   graduates are= welcome to apply.
   

   
Located  in upstate NY, this large university-based= medical center
   is  looking for talented histotechs who can work a flexible= day shift
   (starting at 5:00am, or 6am or 7am -working 81/2 hours shift= with 1/2
   hr  for  lunch) to join their dynamic team. If you are reliable,= take
   pride  in  quality  work,  are  dedicated  to patient care, and have a
   desire=  to  join  an  organization  that values those qualities, this
   might be the= career step you are looking for.
   

   
The  ideal  candidates=  will  have  an  Associate degree in a life
   science  or  higher and hold HT or= HTL ASCP certification. Our client
   will   also   consider   experienced   MTs  that=  are  interested  in
   transitioning  to histology. All candidates must either= have their NY
   license or be eligible for NY licensure.
   

   
Special   features   about   this   position:   Our  client=  offer
   significant   opportunities   for   advancement  and  has  outstanding
   benefits.
   

   
If=  you  meet these qualifications and would like to be considered
   forthis=opportunity,pleasecontact   Darcy   Bloch   at
   [1]dar...@slonepartners.com.
   

   
If  you  have experience= in the diagnostic laboratory industry and
   wish  to be considered for other= roles, please forward your resume to
   Tara Kochis at [2]t...@slonepartners.com.
   

   
All inquiries are kept confidential.

References

   1. 3D"mailto:dar...@slonepartners.com";
   2. 3D"mailto:t...@slonepartners.com";
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[Histonet] Seeking a Histology Manager in CT

[Therese Cook]

   Slone  Partners=  seeks  a  Histology  Manager  for our hospital based
   laboratory client in Connecticut.
   

   
The  successful  candidate  will  have  experience  with  workflow   redesign  
and be great at managing change. The manager will oversee 38
   people,=  including  2 supervisors. Lean and Six Sigma experience is a
   plus.
   = 

   
Qualified  candidates  will  have  a B.S. degree and either HT/HTL   
certification,  with  5 years of supervisory experience, preferably in
   a= busy, high-volume laboratory.
   

   
Special features= of this position: The histology manager will have
   the opportunity= to help redesign this busy laboratory.
   

   
If=  you  meet these qualifications and would like to be considered
   for  this=  position,  please  submit  your  resume to Therese Cook at
   [1]there...@slonepartners.com.
   

   
If  you  have experience in the diagnostic laboratory industry= and
   wish  to be considered for other roles, please forward your resume to   Tara 
Kochis at [2]t...@slonepartners.com.
   

   
All inquiries are kept confidential.

References

   1. 3D"mailto:there...@slonepartners.com";
   2. 3D"mailto:t...@slonepartners.com";
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Re: [Histonet] Immuno. confusion

[Rene J Buesa]

If you fix with formaldehyde, HIER will eliminate the cross link and that step 
is necessary BEFORE you can apply the peroxide. The enzyme (peroxidase) is also 
a protein and has to be "open" to receive the H202.
The same if you want to apply enzymatic digestion, HIER has to be done first.
HIER will remove the formaldehyde cross linking and this step is required 
before any other in the IHC protocol.
René J.

--- On Wed, 8/3/11, Hannen, Valerie  wrote:


From: Hannen, Valerie 
Subject: [Histonet] Immuno. confusion
To: "histonet@lists.utsouthwestern.edu" 
Date: Wednesday, August 3, 2011, 11:57 AM


Hi folks...

I am going to try to revamp out Immuno. procedure and am looking at a couple of 
spec. sheets from our primary vendor. I am gettin conflicting information...so 
I am turning to you all for help.

When doing immuno's that require either HIER or enzyme digestion, do you 
perform the retrieval first or do you do the "peroxide" step first??
One spec. sheets says retrieve then block endogenous peroxidase...the other 
says block endogenous peroxidase then do the HIER or enzyme digestion..which is 
correct??

One other question that I have is...what do you use to enhance your stain?? The 
enhancing solution that we were using has been discontinued.

Thanks so much!!

Valerie Hannen,MLT(ASCP), HTL,SU(FL)
Histotechnologist
Parrish Medical Center
951 N. Washington Avenue
Titusville, Florida 32796
(321) 268-6111 ext. 7506



**
"This email is intended solely for the use of the individual to
whom it is addressed and may contain information that is
privileged, confidential or otherwise exempt from disclosure
under applicable law. If the reader of this email is not the
intended recipient or the employee or agent responsible for
delivering the message to the intended recipient, you are
hereby notified that any dissemination, distribution, or
copying of this communication is strictly prohibited. If you
have received this communication in error, please immediately
delete this message. Thank you"
**
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[Histonet] Immuno. confusion

[Hannen, Valerie]

Hi folks...

I am going to try to revamp out Immuno. procedure and am looking at a couple of 
spec. sheets from our primary vendor. I am gettin conflicting information...so 
I am turning to you all for help.

When doing immuno's that require either HIER or enzyme digestion, do you 
perform the retrieval first or do you do the "peroxide" step first??
One spec. sheets says retrieve then block endogenous peroxidase...the other 
says block endogenous peroxidase then do the HIER or enzyme digestion..which is 
correct??

One other question that I have is...what do you use to enhance your stain?? The 
enhancing solution that we were using has been discontinued.

Thanks so much!!

Valerie Hannen,MLT(ASCP), HTL,SU(FL)
Histotechnologist
Parrish Medical Center
951 N. Washington Avenue
Titusville, Florida 32796
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Re: [Histonet] Processing of derm specimens

[Nicole Tatum]

How many 100%  do you have.  I have 3. This is where the dehydration comes
in. The 100 takes all the mositure out of the specimen. So this step is
critical. Water and xylene are not soluable so if the specimens are not
getting dehydrated properly, the xylene will not penertate the specimens
either. Next is make sure your specimen are not thicker then 3mm.
Espceially lipomas and cyst. Try to cut them as thin as possible. Note,
the caseous inside of a cyst will likely not process and usually expoldes
on a water bath. Next, make sure the specimen cassettes are propely placed
inside the tissue rack. If flow between cassettes is restricted, a portion
of a specimen could be raw.

Hope this helps.

Nicole Tatum HT ASCP

I have recently had a problem with my skin specimens being
> "underprocessed". I use a Leica 300ASP. The schedule is as follows:
> 10% NBF x 2 for 1 hour ea.
> 80% Reagent Alcohol for 1 hour
> 95% Reagent Alcohol x 2 for 1 hour each
> 100% Reagent Alcohol for 1 hour each
> Xylene x 3 for 1 hour each
> Paraffin x 3 for 1 hour each
>
> The specimens are "mushy and swell on the ice"
> Any input is welcome.
>
> send response to :
> _scrochiere@nedlc.com_ (mailto:scrochi...@nedlc.com)
>
> thanks
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Re: [Histonet] paraffin temps

[Rene J Buesa]

The brand of the paraffin has nothing to do with the melting temperature, which 
depends on the paraffin polymers it contains.
Each paraffin will have a melting point value in the package and will melt at 
that temperature. You can use a higher temperature also but not a lower one.
René J.

--- On Wed, 8/3/11, Kolman, Kimberly D.  wrote:


From: Kolman, Kimberly D. 
Subject: [Histonet] paraffin temps
To: histonet@lists.utsouthwestern.edu
Date: Wednesday, August 3, 2011, 8:57 AM


How do you all handle adjustments in paraffin temps depending on
different brands of paraffins? 



Last time I ordered, I received a different brand of Paraplast than my
usual and it is obvious that the temp on the tissue processor needs to
be adjusted up, as the paraffin is collecting in large amounts all over
the top of the cassette basket and is difficult to clean.  To adjust the
temps on the paraffin baths works just fine (as in up to 60 degrees),
which raises the actual paraffin temps up to 62-64 degrees and is then
back to easier melting and cleaning, but the manufacturer packaging
states 'do not heat over 62 degrees'.   



I can revise my QC maintenance sheets to alter the temperature ranges so
I am not constantly out of range; however there is the issue of the
packaging



Can't wait to get all this brand used up; I want my McCormick back!



Kimberly D. Kolman, HT, (ASCP)

Diagnostics 115 

VA Eastern Kansas Health Care System

4101 S. 4th St. Trfwy.

Leavenworth, KS 66048

ph: 913-682-2000 x 52537/52539





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Re: [Histonet] Reorienting frozen tissue

[Rene J Buesa]

Thaw and re-embed in OCT as desired. Freeze again.
René J.

--- On Tue, 8/2/11, Daniela Bodemer  wrote:


From: Daniela Bodemer 
Subject: [Histonet] Reorienting frozen tissue
To: histonet@lists.utsouthwestern.edu
Date: Tuesday, August 2, 2011, 7:44 PM


Hi all,



I would like to know if it is possible and how to reorient or reembedd
human tissue, that has been frozen at -80C in OCT. 



Many thanks,



Daniela Bodemer 

Research Assistant

Surgical Research, Infection and Immunity



Murdoch Childrens Research Institute

The Royal Children's Hospital

Flemington Road Parkville Victoria 3052 Australia 

T 03 9345 5930     T (03 9345 4116)     

E daniela.bode...@mcri.edu.au  

www.mcri.edu.au  



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[Histonet] paraffin temps

[Kolman, Kimberly D.]

How do you all handle adjustments in paraffin temps depending on
different brands of paraffins? 

 

Last time I ordered, I received a different brand of Paraplast than my
usual and it is obvious that the temp on the tissue processor needs to
be adjusted up, as the paraffin is collecting in large amounts all over
the top of the cassette basket and is difficult to clean.  To adjust the
temps on the paraffin baths works just fine (as in up to 60 degrees),
which raises the actual paraffin temps up to 62-64 degrees and is then
back to easier melting and cleaning, but the manufacturer packaging
states 'do not heat over 62 degrees'.   

 

I can revise my QC maintenance sheets to alter the temperature ranges so
I am not constantly out of range; however there is the issue of the
packaging

 

Can't wait to get all this brand used up; I want my McCormick back!

 

Kimberly D. Kolman, HT, (ASCP)

Diagnostics 115 

VA Eastern Kansas Health Care System

4101 S. 4th St. Trfwy.

Leavenworth, KS 66048

ph: 913-682-2000 x 52537/52539

 

 

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Re: [Histonet] Reorienting frozen tissue

[Kim Merriam]

Whenever I have to reembed an OCT block, I leave it out at RT for just a couple 
of minutes to let it that a little bit.  Just take a razor blade and cut out 
the tissue (leaving some OCT around it); re-oreint it in a mold with RT OCT and 
then freeze it.  Don't freeze it too fast, you want the old OCT to melt a 
little bit with the new OCT, that way you will have a block without any seams 
in it.
 
Good luck!
Kim

Kim Merriam, MA, HT(ASCP)QIHC
Cambridge, MA

From: Daniela Bodemer 
To: histonet@lists.utsouthwestern.edu
Sent: Tuesday, August 2, 2011 7:44 PM
Subject: [Histonet] Reorienting frozen tissue

Hi all,



I would like to know if it is possible and how to reorient or reembedd
human tissue, that has been frozen at -80C in OCT. 



Many thanks,



Daniela Bodemer 

Research Assistant

Surgical Research, Infection and Immunity



Murdoch Childrens Research Institute

The Royal Children's Hospital

Flemington Road Parkville Victoria 3052 Australia 

T 03 9345 5930    T (03 9345 4116)    

E daniela.bode...@mcri.edu.au  

www.mcri.edu.au  



This e-mail and any attachments to it (the "Communication") are, unless
otherwise stated, confidential, may contain copyright material and is
for the use only of the intended recipient. If you receive the
Communication in error, please notify the sender immediately by return
e-mail, delete the Communication and the return e-mail, and do not read,
copy, retransmit or otherwise deal with it. Any views expressed in the
Communication are those of the individual sender only, unless expressly
stated to be those of Murdoch Childrens Research Institute (MCRI) ABN 21
006 566 972 or any of its related entities. MCRI does not accept
liability in connection with the integrity of or errors in the
Communication, computer virus, data corruption, interference or delay
arising from or in respect of the Communication. 

P      Please consider the environment before printing this email




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[Histonet] RE: Perforin Ab

[Mauger, Joanne]

Hi Luis,
We use perforin from Thermo-Fisher, #MS-1834.
Jo Mauger
-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Chiriboga, Luis
Sent: Tuesday, August 02, 2011 3:54 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Perforin Ab

Can anyone recommend a perforin ab for use in FFPE human tissue?
Thanks in advance
Luis


Luis Chiriboga Ph.D
OCS Experimental Pathology IHC Core Lab
Bellevue Hospital Center
Department of Pathology 4w27
(212) 562-4667
luis.chirib...@nyumc.org






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