RE: [Histonet] Microtome knives

[Jack Ratliff]

I might also add that Delaware Diamond Knives (DDK) also sells and sharpens 
microtome knives!
Jack

Jack L RatliffRatliff Histology Consultants, LLC317-281-1975



> From: pru...@ihctech.net
> To: max_histo...@yahoo.it; jkr...@deltacollege.edu; 
> Histonet@lists.utsouthwestern.edu
> Date: Sun, 18 Nov 2012 07:50:11 -0700
> Subject: RE: [Histonet] Microtome knives
> CC: 
> 
> There are knife sharpening services Sturkey and Dorn and Hart are two that
> come to mind.  You can also find refurbished disposable blade holders and
> buy disposable blades, the blade holder you get will determine if you use
> low profile or high profile blades.
> 
> Patsy Ruegg, HT(ASCP)QIHC
> Ruegg IHC Consulting, LLC
> 40864 Arkansas Ave
> Bennett, CO 80102
> Phone: 303-644-4538
> Fax: 720-859-4110
> pru...@ihctech.net
> 
> -Original Message-
> From: histonet-boun...@lists.utsouthwestern.edu
> [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Massimo
> Sent: Sunday, November 11, 2012 2:45 AM
> To: Jon Krupp; Histonet@lists.utsouthwestern.edu
> Subject: Re: [Histonet] Microtome knives
> 
> I prefer to sharpen my microtome knives by myself by hand.
> I have a vintage Cambridge Rocking Microtome and despite its age it works
> very well.
> Sharpening is a time consuming for the first time, it's depends on the
> conditions of the blade edge.
> Once you have a nice cutting profile its maintenance it's quite easy and it
> takes a few minutes by
> stroking the knife on a flat glass with oil and a bit of aluminium oxide
> powder (3 -1 micron grits).
> For me sharpening and honing of a microtome knife has became a secondary
> "hobby".
> A solid knife has the advantage, compared to a disposable blade, to be
> liable to less vibrations.
> 
> Kind Regards,
> Massimo Tosi
> 
> 
> "A humble Chemical
> Engineer who loves Histology"
> 
> 
> 
> 
> 
> 
>  Da: Jon Krupp 
> A: Histonet@lists.utsouthwestern.edu 
> Inviato: Venerdì 9 Novembre 2012 19:49
> Oggetto: [Histonet] Microtome knives
>  
> Greetings
> 
> I need some advice regarding microtome knives. I am not  histo tech, I did
> all my sectioning in a plant research lab, but now I find myself needing to
> learn more about histo type methods.
> 
> We have microtomes, AO 820's, and we have a bunch of donated knives. I need
> advice about whether it would be better to find a knife sharpener and use
> the microtome knives we have, or check into getting a disposable knife
> holder. 
> 
> When I was sectioning, we just used a simple razor blade holder. Now I see
> references to high profile and low profile blades and holders, and I don't
> know the difference. 
> 
> Anyone willing to help me out?
> 
> Thanks
> 
> Jon
> 
> Jonathan Krupp
> Delta College
> 5151 Pacific Ave.
> Box 212
> Stockton, CA  95207
> 209-954-5284
> jkr...@deltacollege.edu
> 
> Find us on Facebook @
> Electron Microscopy at SJ Delta College
> 
> 
> 
> 
> 
> 
> 
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[Histonet] CRYOSTAT PROBLEM

[Lakshmi Kammili]

Hi all,
We have a old cryostat (Microm h505 model). It was making some sudden 
vibrating noises for a little while and then smooths it self. There  are no 
temperature alterations every thing was fine but suddenly it stopped working, 
no display more looks like dead :( 
I tried calling the service company which serviced before but I guess the 
company is no more in service (as it was a very old machine). I am very new to 
this maintenance issues , I would be glad if any one of you can help me in 
suggesting a company or person whom I can talk about the issue or some one who 
can come and look at the  instrument and suggest the best advice . I assume 
probably some electric motor is dead inside. 
Any advice will be really helpful.
Thank you,
Lakshmi,
Sr. research assistant,
Pathology core Lab,
George Washington University,
Washington, DC.

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[Histonet] Re: Radiation safety

[Bob Richmond]

Deloris Carter (where?) asks:

>>I'm trying to complete a protocol for radioactive specimen handling.  I
need to know if anyone has suggestions for this.  We do not use a geiger
counter for specimens.  The radiation safety person in nuclear med says
their policy is that technicium 99 has such a short half life, and that
there is no need for special handling of sentinel node specimens.  I just
need some input.  I'm not really getting anywhere with nuclear med on any
other information such as prostate seeds, the actual breast biopsy that may
be sent after a sentinel node procedure, etc.  Our CAP inspection looms,
and I need to finalize this protocol ASAP.  Thanks for any help.<<

Technetium 99m has a half-life of six hours, decaying (emitting a
gamma particle) into technetium 99 which has a half-life of over
100,000 years. The amount of radioactive material in a sentinel node
or a lumpectomy specimen is so small that no special precautions are
needed in handling and processing it. There are a number of good
articles about this available - I can probably find some references -
but this matter was put to rest about 15 years ago.

The radioisotopes in prostate "seeds" are more hazardous. There are
three such isotopes, if all of them are still in use. The
longest-lived has a half-life of 73 days, so it takes about two years
for it to decay to a reasonably safe level.

I've received "seeds" in prostatectomy (TURP) specimens, with no
information as to what they were or how old they were. Although they
are probably not very hazardous, I consider this to be serious
negligence. Here's what they look like, from a specimen I had several
years ago (the "seeds" were four years old, I finally found out).
http://www.flickr.com/photos/bobrichmond/5833004125/in/set-72157618450128961

I would suggest not processing the specimen (or opening the container)
until adequate information is obtained. Expect to be treated with
great condescension when you inquire. I don't know of any references
on the subject.

I don't know how CAP addresses the problem.

Bob Richmond
Samurai Pathologist
Maryville TN

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[Histonet] Re: Lung histology

[Erickson, Jamie E]

HI  Zakky,
   I've worked on a lot on mouse lung and the process of 
embedding can depend on what you are looking for or your pathologist opinion.
In my OVA models  I was looking at inflammation around the main stem bronchi 
and looking at goblet cell hyperplasia  using PAS stain. Because we used an 
intranasal  route of administration we wanted to look to see if there was a 
difference in PAS at the main stem bronchi compared to  distal secondary and 
tertiary bronchi so we chose to embed sagittal.
I isolated the left lobe that is the lobe with 
no accessory lobes then trimmed in  to the main stem but not to close,  this 
was done to make a flat surface for embedding. Then you can cut into the lung 
until you get a nice full or almost full length of the main stem bronchi. I did 
PAS on these and image analysis to quantitate amount of PAS production. 
Perfusion of the lungs was a must for this view.
There are lots of papers that do this by cross sections as well and stain with 
collagen deposition stains like sirus red, so you'll have to see what fits your 
study.
Whatever you choose be consistent with your location of sectioning, so you can 
compare samples across groups that are taken from the same anatomical location.

Hope that helps  Good luck,

Jamie Erickson (ASCP) HTL
Pharmacology Dept.
Abbott laboratories
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[Histonet] Re: Histonet Digest, Vol 108, Issue 34

[Madeleine Huey]

On Wed, Nov 28, 2012 at 7:10 AM,
 wrote:
> From: histonet-boun...@lists.utsouthwestern.edu 
> [histonet-boun...@lists.utsouthwestern.edu] on behalf of Kim Merriam 
> [kmerriam2...@yahoo.com]
> Sent: Wednesday, November 28, 2012 7:54 AM
> To: Histonet
> Subject: [Histonet] eosin in the processor
>
> Hi Everyone,
>
> Years ago, my lab used to put eosin in the processor to lightly tint the 
> smaller mouse tissues.  I can't remember which station we put it in (I think 
> it was the 2nd 100% ethanol).  Also, back then my lab didn't do any IHC; will 
> the eosin affect any IHC that might be done (I am guessing no, but I want to 
> be sure).
>
> Thanks,
> Kim
>
> Kim Merriam, MA, HT(ASCP)QIHC
> Cambridge, MA

Hello Kim,
I always put the Eosin in the last two 100% alcohol.  The whole
concepts is to have your tissues pick up the Eosin and not wash out
during the processing. Therefore if you put them in the beginning
(ie.70%, 85% ) and not the 100% alcohol, they will eventually be
wash out.
Madeleine Huey BS, HTL & QIHC (ASCP)
Supervisor - Pathology (IPOX & Histology)
madelein...@elcaminohospital.org

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[Histonet] Re: Histonet Digest, Vol 108, Issue 37

[Madeleine Huey]

Message: 2
Date: Thu, 29 Nov 2012 16:46:03 +
From: zakky cholisoh 
Subject: [Histonet] Lung histology
To: histonet@lists.utsouthwestern.edu
Message-ID:

Content-Type: text/plain; charset=ISO-8859-1

Hello lovely people!

Is there anyone working with mouse lung (or any lung)?
I work with respiratory virus in mice and now about to begin the fun part
of the work, histopathology :)
I was just wondering what is the best section of the lung, is it coronal
section or tranversal (horizontal) section?

Any advice?

Thanks so much in advance

Zakky
PhD student
Respiratory Pharmacology
Welsh School of Pharmacy

Hello Zakky,
It all depends!  One of my previous project  I am screening tumors
from the mouse lungs, therefore I always do them on Tranversal
(seperate all 5 lobes first, then embed horizontal).  The thinner the
block (horizontal) the lesser sections I need to cut.
Hope this answer your questions.
Madeleine Huey BS, HTL & QIHC (ASCP)
Supervisor - Pathology (IPOX & Histology)
madelein...@elcaminohospital.org

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[Histonet] cellblock help

[Davis, Cassie]

Good afternoon Histonet folks,
 I am hoping somebody might be able to help me with a protocol using Thrombin 
to get a cellblock from a specimen with scant material.
This process was brought to my attention a few years ago but I never saw an 
actual protocol. Thanks for any help & have an awesome day.
Cassandra Davis
cda...@che-east.org
302-575-8095


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RE: [Histonet] Radiatiation Safety

[Craven Peter (NHS HIGHLAND)]

Deloris
When we set this up we got our Radiation Protection Services to write a safe 
handling policy for the tissues (sentinel nodes) and added this into our 
Standard Operating Procedures. Might as well get the advice from the experts 
and trust it rather than making your own policy.

Peter

Peter L Craven FIBMS
Pathology Department
Raigmore Hospital
Old Perth Road
Inverness
IV2 3UJ
Tel 01463 704269
email  peter.cra...@nhs.net

From: histonet-boun...@lists.utsouthwestern.edu 
[histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Deloris Carter 
[dels...@gmail.com]
Sent: 29 November 2012 04:59 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Radiatiation Safety

Hi all,

 I'm trying to complete a protocol for radioactive specimen handling.  I
need to know if anyone has suggestions for this.  We do not use a geiger
counter for specimens.  The radiation safety person in nuclear med says
their policy is that technicium 99 has such a short half life, and that
there is no need for special handling of sentinel node specimens.  I just
need some input.  I'm not really getting anywhere with nuclear med on any
other information such as prostate seeds, the actual breast biopsy that may
be sent after a sentinel node procedure, etc.  Our CAP inspection looms,
and I need to finalize this protocol ASAP.  Thanks for any help.

Deloris Carter HT(ASCP)
dels...@gmail.com
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[Histonet] Radiatiation Safety

[Deloris Carter]

Hi all,

 I'm trying to complete a protocol for radioactive specimen handling.  I
need to know if anyone has suggestions for this.  We do not use a geiger
counter for specimens.  The radiation safety person in nuclear med says
their policy is that technicium 99 has such a short half life, and that
there is no need for special handling of sentinel node specimens.  I just
need some input.  I'm not really getting anywhere with nuclear med on any
other information such as prostate seeds, the actual breast biopsy that may
be sent after a sentinel node procedure, etc.  Our CAP inspection looms,
and I need to finalize this protocol ASAP.  Thanks for any help.

Deloris Carter HT(ASCP)
dels...@gmail.com
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[Histonet] Lung histology

[zakky cholisoh]

Hello lovely people!

Is there anyone working with mouse lung (or any lung)?
I work with respiratory virus in mice and now about to begin the fun part
of the work, histopathology :)
I was just wondering what is the best section of the lung, is it coronal
section or tranversal (horizontal) section?

Any advice?

Thanks so much in advance

Zakky
PhD student
Respiratory Pharmacology
Welsh School of Pharmacy
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[Histonet] Hii everybody

[vaigunda ragavendran]

Hii Histonetters,

Happy holidays. I am on the lookout for a marker to be checked using 
immunohistochemistry that would get expressed at the DRG in response to 
streptozotocin-induced diabetes. I am basically looking for a neuronal marker 
that would give an idea of the degree of microvascular dysfunction induced by 
diabetes. Sounds very complicated though. Any suggestions are most welcome.


Thanks and bye.

Rags,

Postdoctoral Research Assistant,
Coderre Lab,
Department of Anesthesia,
Room No: 1201, McIntyre Medical Building,
3655, Promenade Sir William Osler,
Montreal, Quebec H3G 1Y6,
Canada.

Phone (Lab): +1 514-398-2903
(Facsimile): +1 514-398-8241

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Re: [Histonet] Validation of a new tissue processor

[Rene J Buesa]

Absolutely agree with Tim's comments. 
I would even add that if the new tissue processor uses the "standard" 
heat/agitation/vacuum/pressue sequences as the old one, the validation 
objective is mainly directed to have some documentation that the facility's 
attorneys can have in case "something goes wrong" and the plaintiff's attorney 
asks: "Was the instrument validated?" avoiding that a judge, also ignorant of 
our methods, rules against our lab.
The validation is really necessary if the processing technology goes from the 
"conventional" to the microwave technology where the times are so different and 
the development of new quality protocols are of paramount importance. 
The one who has to decide about the validation test is the pathologist and his 
acceptance has to go to the QC file, and that's it!
René J. 

From: "Morken, Timothy" 
To: "'histonet@lists.utsouthwestern.edu'"  
Sent: Wednesday, November 28, 2012 5:10 PM
Subject: RE: [Histonet] Validation of a new tissue processor

I don't think it is necessary to test every single stain or antibody you do to 
validate a new tissue processor. Most of the time the only thing changing is 
the machine, not the chemicals you use. The point of validation is to prove 
that the system gives you the same results as the old instrument. Using a 
representative sample of types of stains and antibodies will give you 
confidence that it works fine and save a huge amount of money and time. 

Think about it from this perspective: We get paraffin slides and blocks from 
consult cases from all over the place, processed on many different machines 
with infinitely variable time and chemical protocols. Yet virtually all the 
cases work fine in our immuno staining system. It is rare to find cases that do 
not. That alone tells you that massive validation is not really necessary.

The point of a validation procedure is to plan one that you feel will give you 
confidence that the new instrument gives you the same results as the old one. 
If you do insist on testing "all" your antibodies (we have over 200 here) then 
tissue arrays or multiple tissue blocks will save a lot of time and money. 

Of course, if you go to a new processor that uses totally different  types of 
chemicals and methods then validation of every antibody may be necessary. Even 
then, however, representative markers of various types may suffice. 

Tim Morken
Department of Pathology
UC San Francisco Medical Center


-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Helen Fedor
Sent: Wednesday, November 28, 2012 1:51 PM
To: 'Rene J Buesa'; Howe, Helena; 'histonet@lists.utsouthwestern.edu'
Subject: RE: [Histonet] Validation of a new tissue processor

You will also need to validate all of the IHC stains and special stains that 
you do on these tissues. You can take samples from each tissue type and run 
them in parallel on both machines. Then test all of your special stains and 
antibodies that you have in your inventory.

:)
Helen

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa
Sent: Wednesday, November 28, 2012 4:47 PM
To: Howe, Helena; 'histonet@lists.utsouthwestern.edu'
Subject: Re: [Histonet] Validation of a new tissue processor

Using at least 25 different types of tissues, prepare parallel almost identical 
pairs of slices for each tissue and process them simultaneously in your new 
tissue processor and in the old one.
Prepare H&E sections and give them to the pathologists concealing the used 
tissue processor. The pathologists should decide if there are differences 
between the pairs of sections.
The pathologist's opinion should be registered in the QC file and that will be 
your validation.
René J.

From: "Howe, Helena" 
To: "'histonet@lists.utsouthwestern.edu'"  
Sent: Wednesday, November 28, 2012 2:57 PM
Subject: [Histonet] Validation of a new tissue processor

Does anyone know what the CAP is requiring to validate a new tissue processor?
Thank You
Helena Howe
ProMedica Laboratories
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[Histonet] RE: My earlier post about ASCP requirements in MI

[Pam Barker]

Hi Histonetters!!
Thank you to all my histobuddies that responded about the certification in
Michigan question. Of course I am going to encourage them to hire a
certified tech it was a question the client asked that I needed to get
answered and I appreciate you taking the time. Happy Holidays!!

Happy Holidays !!

Thank You!
  
 
Pam Barker
President/Senior Recruiting Specialist-Histology
RELIA Solutions
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.com  /PamBarkerRELIA
www.linkedin.com/in/reliasolutions
www.twitter.com/pamatrelia 


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Re: [Histonet] chemical disposal

[Lee & Peggy Wenk]

Where most people get confused is the"Biological Hazard". They think - this 
chemical would hurt a human being, it would damage someone's skin if 
splashed on it, it would injure someone's lungs if inhaled, it could cause 
cancer with long exposure, etc. Since it's hurting a person, it must be a 
biological hazard. Nope.


"Biological hazard" means that the "solution" contains a biologically active 
microorganism (bacteria, fungus, virus, parasite) or a prion - all of which 
could infect the person using the "solution" if not handled correctly. 
(Fresh tissue would be a possible biological hazard, since we don't know 
what, if anything, that tissue is infected with. Same with a frozen section. 
If tissue is properly fixed and processed, that would kill the 
microorganism, so the tissue is no longer a biological hazard. Prions need 
special treatment with formic acid.)


If the material is a chemical (solvent, dye, bleach, soap, toner cartridges 
that leak, etc.), then it is a Chemical Hazard. This would include the DAB.


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

The opinions expressed are mine, and do not reflect on Beaumont Hospital.

-Original Message- 
From: Cynthia Pyse

Sent: Wednesday, November 28, 2012 3:44 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] chemical disposal

Quick question for Histoland. I am having a debate about DAB disposal. Our
general manager ( non lab background) insists that the liquid DAB can go
into a biological hazardous waste. I disagree, it is a chemical and needs to
be disposed in the chemical hazardous waste. What is everyone else doing to
dispose of DAB. We are located in NY, I do have those regs. Thanks in
advance for any and all help.

Cindy



Cindy Pyse, CLT, HT (ASCP)

Laboratory Manager

X-Cell Laboratories

20 Northpointe Parkway Suite 100

Amherst, NY 14228

716-250-9235 etx. 232

e-mail cp...@x-celllab.com



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