[Histonet] No Frost Freezers !

[Cesar Francisco Romero]

 

I haven’t seen any answer to the question below, and I
am curious about it. 

César Romero

Buenos Aires

Argentina 

Message: 19

Date: Wed, 18 Feb 2015 22:31:54 +

From: James Watson 

Subject: [Histonet] Storing antibodies in a frostless freezer

To: "'histonet@lists.utsouthwestern.edu'"



Message-ID:



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



Being old school I was taught that antibodies do not get stored in a frostless
freezer.  I am now being told that the
new frostless freezers do not go above -13°C during the defrost cycle and it is
safe to use.  Teach this old dog a new
trick, is anyone using a frostless freezer for storing antibodies?



Jamie



James Watson HT  ASCP

GNF  Genomics Institute of the Novartis
Research Foundation

Scientific Technical Leader II, Histology

Tel858-332-4647

Fax  
858-812-1915

jwat...@gnf.org



  
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[Histonet] RE: Know Error

[Cesar Francisco Romero]

 

I live in a Corrupt Country . 

I can assure to you that this request is simply that,
Corruption. The well done job is not well paid. The one that returns money to a 
corrupt person or a group of persons , yes, it is well paid.
 César RomeroBuenos AiresArgentina

  
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[Histonet] Re: Decalcified Bone Staining Problems.

[Cesar Francisco Romero]

 

In the link
below ( Figure 5 ) Incomplete decalcified Bone: Basophilic and Brittle.



http://www.dentalaegis.com/cced/2011/03/alveolar-ridge-augmentation-comparion-of-two-socket-graft-materials-in-implant-cases

 


In the link below Complete Decalcified Bone: Pink and with well preserved 
morphology.

http://www.visualphotos.com/image/1x8802590/decalcified_compact_bones Cesar 
RomeroBuenos AiresArgentina

  
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[Histonet] Re: Decalcified Bone Eosin Overstaining

[Cesar Francisco Romero]

 

In
the past I have been consulted for lost of DAB staining in IHC when using acid
alcohol to differentiate hematoxylin.

In
my Latitude tap water is enough for bluing hematoxylin but I think that you
need something else.Trizma Base is a good bluing agent when tap water is not 
good enough.Just put a spoon of trizma in water and see the results.
Cesar RomeroBuenos AiresArgentina

  
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[Histonet] Re: Papillary Vs Reticular Dermis.

[Cesar Francisco Romero]

 

Another Hallmark is the Superficial Vascular Plexus of
the Skin. 

You can do Muscle Actin ( HHF – 35 ) or CD 34 to
remark the vessels. 

Link to a Picture 

http://biologiedelapeau.fr/spip.php?page=forum&id_article=21&lang=en 

This Superficial Vascular Plexus allows separating
Invasive Melanoma Clark´s Level III from Level IV.

  
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[Histonet] RE: Papillary Vs Reticular Dermis

[Cesar Francisco Romero]

 

A good trichrome stain can help very much.

Here is a link to a picture.

http://download.videohelp.com/vitualis/med/his_pic_integument2.htm

  
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[Histonet] RE: Eosin Shades

[Cesar Francisco Romero]

 
   

I think that the nearest to what you wish is Eosin
- Phloxine Stain.

Here is the web page where you can find the formula.

http://protocolsonline.com/histology/dyes-and-stains/haematoxylin-eosin-he-staining/

 

I use it with very good results.

   

  
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[Histonet] RE: Decalcifying and Processing large bones

[Cesar Francisco Romero]

For many years now I have used this protocol in Bone
Marrow samples.

The cortical of the bone is still a problem, but it
can be solved with longer decal time.

If you need to preserve morphology and antigenicity
for Immunohistochemistry this is the best choice.

Three Days Bouin’s Fixative ( From the very Beginning
), don’t use Formalin.

Three Days of a Saturated Solution of EDTA ( Disodium
Salt ) in distilled water.

After this time put the blocks in the first alcohol and
process as usual.

Don’t put the blocks in formalin.

EDTA alone or with Heating  affects morphology very badly.

For IHC Antigen Retrieval is the recommended or used
for each antibody.

Difficult ones as CD 5 or CD 23 work very well with
this decal protocol.

Cesar
Romero

Buenos
Aires.

Argentina.

  
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[Histonet] Mineral Oil in Microwave Processing.

[Cesar Francisco Romero]

Dear People from
Histonet


Do you still use
Mineral Oil in your Microwave Processing Protocol ?


Or you only use
ethyl alcohol ~ Isopropyl alcohol and Paraffin .


I will apreciate
any advice.


Thank you all


Cesar Romero


Buenos Aires


Argentina


  
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[Histonet] RE: Rush Processing Protocol

[Cesar Francisco Romero]

I
think that what you really need is a Microwave Processor.

But
I live in a Poor World with No Machines.

I
have to report small wedge Kidney Biopsies for transplantation in two hours .

This
is my Rush Processing Protocol:

Formalin
( 1 : 10 ) 10 minutes ( they already come from the surgery room in formalin ).

Ethyl
Alcohol 96° 10 minutes.

Ethyl
Alcohol 100 ° 10 minutes.

Ethyl
Alcohol 100 ° 10 minutes.

Xylene 15
minutes.

Paraffin 15
to 20 minutes.

The secret
is that I already have all the containers with the reagents in the stove at 62°
C and I do all the processing inside the stove. I hope it helps. César 
RomeroBuenos AiresArgentina

  
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[Histonet] RE: Reticulum Vs Reticulin

[Cesar Francisco Romero]

Reticulum
means Net ( Red in Spanish or Web in English ).

It refers
to the Net that exist in Lymphoid tissue of any kind ( Payer Patches – Lymph 
Nodes
– Spleen ).

Dendritic
Cells attach to this Net, that’s why in the past it was used the term Reticulum
Cells Sarcoma ( for Dendritic Cells Sarcoma ).

Reticulin (
Reticulina in Spanish ) is the Silver Technic for Staining this Type III 
Collagen
Fibers of the Net.

  
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[Histonet] RE: Grossing Prostate Biopsies

[Cesar Francisco Romero]

I gross
prostate needle biopsies every week.

I receive
12 samples but convert them in 6 paraffin blocks.

I stain
the Medial Samples of each level and each side with Eosin and the Lateral
Samples with EA - 36 .Example: Base Lateral Right ( EA - 36 ), with Base Medial 
Right ( Eosin ).Base Lateral Left ( EA - 36 ), with Base Medial Left ( Eosin ) 
and so on.

I wash
the excess of colorant with 96 alcohol, wrap the samples of each sextant  in 
one paper and put
them in one cassette.

Then I
put the Cassette in the First Alcohol to avoid washing the Stains with aqueous
formalin.

It is
very important to cut the cylinders so they do not exceed 5 mm. long.  You 
avoid the coiling and facilitate the
embedding.

After all
that it is very easy to see the colors in the paraffin block.It saves time and 
money and there is no errors if you are a tidy quiet person. Cesar RomeroBuenos 
AiresArgentina 

  
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[Histonet] Re: Fixation Question

[Cesar Francisco Romero]

I forgot to
say that the rest of the tissue processing can cause shrinkage.

If the
paraffin is too hot you will get fried tissues.

Here is a
Link with very good information about tissue processing. 

http://users.adam.com.au/royellis/tp/tp.htm

  
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[Histonet] RE: Fixation Question

[Cesar Francisco Romero]

The total Osmolarity of the
fixing solution should be isosmotic with tissue fluids in order to prevent
shrinkage or swelling of cells. 

The best way to avoid shrinkage
is to prepare Formalin with Physiological or Isotonic Saline Solution.

The solution is 9 grams of sodium chloride ( NaCl )
dissolved in Distilled Water, to a total volume of 1000 ml.

To prepare the fixative use 100 ml of pure Formalin (
Formalin 37 ~ 40 % ) and 900 ml of Isotonic Saline Solution.


Cesar Romero
Buenos Aires
Argentina
  
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[Histonet] RE: India Ink for inking surgical margin borders

[Cesar Francisco Romero]

I use Artist
China ( India ) Ink.

The secret
is to immerse the blocks in Bouin’s Fixative to coagulate the Ink.

After that
put the tissue blocks directly in the First Alcohol.

Don’t go
back to formalin.


Cesar Romero
Buenos Aires
Argentina
  
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[Histonet] Myc Protein Antibody and Protocol

[Cesar Francisco Romero]

Dear People
from Histonet

I need to
know if somebody is using Anti Myc Protein Antibody in the Panel for Large Cell
Lymphomas.

I will appreciate
any advice on the brand of the antibody and the protocol to make it work.

I do not
use any Automated Stainer, only Manual Immunohistochemistry.

Thank You
All in advance.

Sincerely yours

Cesar Romero

Buenos Aires

Argentina

  
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[Histonet] RE: Plastic/Plexiglass Incubation chamber for IHC slides

[Cesar Francisco Romero]

For
Manual IHC I suggest best the Sequenza Slide Rack from Thermo Scientific. 
It makes your life easier and you can save money, because you only need 100 ul 
of any reactive per slide.




Here is the link to the web page.

 


http://www.thermo.fi/com/cda/product/detail/0,1055,105630785,00.html

 

I hope it helps.

Cesar Romero

Buenos Aires

Argentina

  
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[Histonet] RE: Histonet Digest, Vol 115, Issue 11

[Cesar Francisco Romero]

Her 2 Antibody
 
I do Manual IHC with Tissue Tek Slide Racks  and have the same problem.
I think the problem is in the holder clip.
Only the top sample stains well.
Cesar Romero
Buenos Aires
Argentina
 
> From: histonet-requ...@lists.utsouthwestern.edu
> Subject: Histonet Digest, Vol 115, Issue 11
> To: histonet@lists.utsouthwestern.edu
> Date: Tue, 11 Jun 2013 10:00:54 -0700
> 
> Send Histonet mailing list submissions to
>   histonet@lists.utsouthwestern.edu
> 
> To subscribe or unsubscribe via the World Wide Web, visit
>   http://lists.utsouthwestern.edu/mailman/listinfo/histonet
> or, via email, send a message with subject or body 'help' to
>   histonet-requ...@lists.utsouthwestern.edu
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>   histonet-ow...@lists.utsouthwestern.edu
> 
> When replying, please edit your Subject line so it is more specific
> than "Re: Contents of Histonet digest..."
> 
> 
> Today's Topics:
> 
>1. RE: samples (Webb, Dorothy L)
>2. Re: Histonet Digest, Vol 115, Issue 10 (Wanda Shotsberger Gray)
>3. RE: bone marrow aspirations (Cathy)
>4. Her-2 antibody (Robert Fauck [CCDHB])
>5. Heinz body stain on already dried blood smears.
>   (Jackman, Mary Ione)
>6. Telepathology (Gloria Tharp)
>7. RE: Telepathology (Elizabeth Chlipala)
> 
> 
> --
> 
> Message: 1
> Date: Mon, 10 Jun 2013 12:24:04 -0500
> From: "Webb, Dorothy L" 
> Subject: [Histonet] RE: samples
> To: "'sa...@royalmarker.com'" 
> Cc: "'histonet@lists.utsouthwestern.edu'"
>   
> Message-ID:
>   <65365f35c0f2ef4d846ec3ca73e49c4302758cf69...@hpemx3.healthpartners.int>
>   
> Content-Type: text/plain; charset="utf-8"
> 
> It is actually not a website but the internet communication (like a 
> listserve) they have for PA’s.  For instance, histology has one at 
> histonet@lists.utsouthwestern.edu 
> that is used to communicate and network online.
> 
> Thanks again!
> 
> Dorothy Webb
> 
> From: sa...@royalmarker.com [mailto:sa...@royalmarker.com]
> Sent: Monday, June 10, 2013 12:20 PM
> To: Webb, Dorothy L
> Subject: RE: samples
> 
> No problem, the samples are on the way. How were you recommended for our 
> product? Could you let us know which website you are referring to?
> 
> Thanks Again,
> Shawn Jenkins
> 
> 
>  Original Message 
> Subject: RE: samples
> From: "Webb, Dorothy L" 
> mailto:dorothy.l.w...@healthpartners.com>>
> Date: Fri, June 07, 2013 1:40 pm
> To: "'sa...@royalmarker.com'" 
> mailto:sa...@royalmarker.com>>
> Thank you very much.  Your marking inks were recommended on the website for 
> PA’s.
> 
> Your efficient response is greatly appreciated and we will get back to you 
> next week with feedback!
> 
> Dorothy Webb
> 
> From: sa...@royalmarker.com 
> [mailto:sa...@royalmarker.com]
> Sent: Friday, June 07, 2013 12:30 PM
> To: Webb, Dorothy L
> Subject: RE: samples
> 
> Hello Dorothy,
> 
> We will gladly send you a sample pack of our marking dyes to try. It will 
> ship out today via UPS to the address below that you provided. I'll be sure 
> to have your name as well as Jesse Spates on the label as well for a contact. 
> The set will include 10 1/4oz bottles (all of our colors). Please contact us 
> with any comments on the product. We appreciate your feedback and hope you 
> are happy with our products. Have a great weekend and look forward to hearing 
> from you.
> 
> Best Regards,
> Shawn Jenkins
> Royal Marker
> (P) 954-462-0264
> (F) 954-374-7290
> 
> 
>  Original Message 
> Subject: samples
> From: "Webb, Dorothy L" 
> mailto:dorothy.l.w...@healthpartners.com>>
> Date: Fri, June 07, 2013 11:59 am
> To: "'sa...@royalmarker.com'" 
> mailto:sa...@royalmarker.com>>
> Cc: "Spates, Jesse D" 
> mailto:jesse.d.spa...@healthpartners.com>>
> Our PA’s are looking to try other inks than what we are currently using so we 
> are wondering if we could please get some ink samples.
> 
> Our hospital is Regions Hospital
>640 Jackson St.
>St. Paul, MN. 55101
> 
> I am the technical specialist of histology, Dorothy Webb and the lead PA is 
> Jesse Spates @ 651-254-4913.
> 
> Thank you,
> 
> Dorothy Webb
> Regions Histology Technical Specialist
> 651-254-2962
> 
> 
>   
> This e-mail and any files transmitted with it are confidential and are 
> intended solely for the use of the individual or entity to whom they are 
> addressed. If you are not the intended recipient or the individual 
> responsible for delivering the e-mail to the intended recipient, please be 
> advised that you have received this e-mail in error and that any use, 
> dissemination, forwarding, printing, or copying of this e-mail is strictly 
> prohibited.
> 
> If you have received thi

[Histonet] Amyloid

[Cesar Francisco Romero]

I think this is a great article to read about amyloid. This is the link to the 
paper 
http://propath.com/newsletters-immunohistochemistry-173/181-september-2004-immunohistochemistry-in-amyloidosis
 It belongs to a great Pathologist Dr. Rodney T. Miller

Dr. César RomeroBuenos AiresArgentina 
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[Histonet] How Have You Been Lately?

[Cesar Francisco Romero]

Hello Dear,
How are you doing these days?
You know, I bought one new pair of Nike shoes on one great online shop 
www.OkIsell.com. They are so amazing!
They provide a lot of other brand new casual shoes, sport shoes, high heels, 
ugg boots, such as Prada, Christian Louboutin, Gucci, Lascote, Puma, and so on. 
The styles are the most popular ones for the 2010 year. Have to say it is a 
great and specialize online shoes store.
Hope you can find some great things there too www.OkIsell.com.!
Give my regards to your family then
  
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[Histonet] C4d Antibody

[Cesar Francisco Romero]

Dear People from Histonet

 

Could you share your experience with C4d Antibody ?

 

I couldn't get any results with the mouse monoclonal from Quidel.

 

I will try now the rabbit polyclonal ( C4dpAb )from Biomedica - Grouppe.

 

I work in a Public Hospital with many kidney transplantations per year.

 

Thank You in advance.

 

César Romero

Buenos Aires

Argentina

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