Re: [Histonet] Dry Film Coverslipping?

[Rene J Buesa via Histonet]

 A Sakura film cover slipper will not work without xylene.Now that you have 
decided to go "xylene free" just go "all the way in" and after staining the 
slides DRY them in an oven at 60ºC during 5 minutes and cover.If you want 
further information on "cover slipping without xylene" send me your e-mail 
address and I will send you a copy of a short paper I published on this 
subject.René 
On Wednesday, October 6, 2021, 01:17:48 PM GMT-4, Cates, Julia via Histonet 
 wrote:  
 
 Hello Histonet,

I have been asked to consider going Xylene free.  The only hang up that I am 
having is the coverslipping part.  We have an old Sakura Film coverslipper and 
I don't know how that would work without xylene.  When I did some searching in 
the archives, I found some comments made by Rene, that he used this type of 
coverslipper on oven dried slides but never referenced how.  Does anyone use 
this method in your lab?  If so, would you share your protocol and are there 
any problems to consider?

Thanks,

Julia C.


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Re: [Histonet] combining cytology and histology

[Rene J Buesa via Histonet]

 Consider the cytology lab as if it were another section of histology, like 
"special stains".Staff it with at least 1 samples processor + 1 
cytotechnologist.Samples should be accessioned separately.René
On Monday, August 9, 2021, 06:03:03 PM GMT-4, Richardson, Pam K via 
Histonet  wrote:  
 
 Does anyone have a lab that has combined histology and cytology? I would like 
to know how they are staffed and if accessioning handles all samples or if they 
are handled separate? 

Best Wishes

Pam ~
National Histology Professionals Day 3/10/21
Pathologists' Assistant Day 4/14/2021
Medical Laboratory Professionals Week April 18-24, 2021
National Cytotechnology Day 5/13/2021

+++
Pam Richardson
Clinical Manager
Gundersen Health System Laboratory Services
Email: pkric...@gundersenhealth.org
Phone: 608 775-4133
Fax: 608 775-6136
Interdepartmental Mail Stop: H04-007
E-visit us at: http://www.gundersenhealth.org

-Original Message-
From: histonet-requ...@lists.utsouthwestern.edu 
 
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To: histonet@lists.utsouthwestern.edu
Subject: Histonet Digest, Vol 212, Issue 17

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Re: [Histonet] Detecting copper in H staining?

[Rene J Buesa via Histonet]

 Unfortunately your pathologist is wrong. You will need the "rhodamine" special 
IHC method to identify copper in tissue sections.René
On Tuesday, September 10, 2019, 11:08:09 AM EDT, Jennifer Phinney via 
Histonet  wrote:  
 
 Hello all,
I am in a veterinary histology lab. One of my Pathologists mentioned that at 
two of the locations he'd been at previously (Ithaca, NY and Saskatoon, Canada) 
copper deposits stained in the routine H slides. Is anyone aware of how this 
could have been achieved? I assume it would either have to be an additive in 
the eosin or perhaps something naturally occurring in the tap water rinses?


Thanks for any assistance,
Jennifer Phinney QIHC
Histology Laboratory Administrator
KSVDL
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Re: [Histonet] Xylene substitutes for clearing agents

[Rene J Buesa via Histonet]

 Under separate cover I am sending you 3 papers of mine that answer your 
question.René
On Wednesday, August 28, 2019, 08:45:31 AM EDT, Ingles Claire via Histonet 
 wrote:  
 
 Chris:
Propar from Anatech works great for us. I believe it is still advisable to use 
xylene in the cleaning cycle on the processors though. We had to go back the 
other way a bit when our Doc wanted a tape coverslipper. Now he gripes about 
the xylene smell. Hmmm.
Claire


From: Hagon, Christopher (Health) via Histonet 

Sent: Tuesday, August 27, 2019 10:53 PM
To: histonet@lists.utsouthwestern.edu 
Subject: [Histonet] Xylene substitutes for clearing agents

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system.
DO NOT CLICK on links or attachments unless you recognize the sender and know 
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UNOFFICIAL

Hello histonetters!

I realise that this has been asked a lot, but cannot find a good link for the 
comparisons of each. I am charged with looking into converting our lab to go 
xylene free. We don't want to go down the limonene path, so that leaves the 
isopropyl alcohol method, or the aliphatic hydrocarbon xylene substitution 
(Leica Sub-X etc).

Looking for opinions from each camp if possible, on how easy or hard it was to 
change procedures. Was there much trial and error in changing the processing 
protocols with the aliphatics? Any pitfalls I should look out for?

Any input greatly appreciated.


Chris Hagon | Senior Scientist, Anatomical Pathology
ACT Pathology | health.act.gov.au



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Re: [Histonet] DAB enhaced with Nickel Sulfate Ammonium

[Rene J Buesa via Histonet]

 Better try another counterstaining. Will be easier in the long run.René
On Monday, August 5, 2019, 01:08:38 PM EDT, Alonso Martínez Canabal via 
Histonet  wrote:  
 
   Good Afternoon.
        It is great to be in this list again. My first e-mail is to ask
your advise about a little problem that we have in the lab.
      We are doing immunohistochemistry in brain tissue using DAB enhanced
with nickel ammonium sulfate. The result is great, with a very clean signal
in 50um thickness brain sections. Our problem is that later we do some
counterstaining with methyl green. It is purified with chorophorm and in pH
4.2 in Acetate buffer. We clearly see the green nuclei and the
immunoreactive cells, but apparently, after the counterstaining, the
dark-purple-blackish precipitate is gone and we can only see regular brown
DAB.
    What do you think?
  I wonder if it is the pH of the Methyl green, but I am kind of lost here.
 Any advise?
    Thank you very much
          Dr. Alonso

-- 
Dr. Alonso Martínez Canabal PhD
Profesor Asociado "C"
Departamento de Biología Celular, Facultad de Ciencias, UNAM
Investigador Nacional "I"
56224833
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Re: [Histonet] Softening Calf Hooves

[Rene J Buesa via Histonet]

Either potassium or sodium hydroxide will work at 10% aq. solution. Check it 
daily until soft enough.As to your "swallow" question, I "Goggled" your 
question and the answer is the following"
In the end, it's concluded that the airspeed velocity of a (European) unladen 
swallow is about 24 miles per hour or 11 meters per second. But, the real 
question is not about swallows at all. King Arthur in the movie had two coconut 
shells that he banged together to simulate the sound of a horse galloping.Jul 
7, 2013 
René
 

On Thursday, March 21, 2019 12:06 PM, Sandra Cheasty via Histonet 
 wrote:
 

 Hello all,
                I have 1 inch thick cross sections (band saw) of calf hooves. 
The pathologist has already decaled the bone, and now we have to soften the 
hooves so she can take thinner sections for processing.


*        Which is better, potassium hydroxide or sodium hydroxide, and what 
concentration?

*        How often should they be checked for softening?

*        What is the relative air speed of an un-laden swallow? (European)

Thanks for your help!
Sandy

Sandra J. Cheasty, HT (ASCP)
Histology & Necropsy Supervisor
UW-Madison, School of Veterinary Medicine

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Re: [Histonet] Stainer vs. Stainer

[Rene J Buesa via Histonet]

Stainers and coverslippers:Let me interject in the discussion:1- Sakura is 
"hands down" my preferred option because of reliability and results 
consistency2- Having to have a water free last ethanol is something of the past 
(besides just proper protocol) on the other hand for years I have dewaxed my 
sections with dishwasher soap and obtained excellent dewaxing without xylene or 
ethanol, at all.3- After staining (or any "staining" protocol) I dehydrate any, 
and I mean any, "stained" by placing them in a 60ºC oven during 5-6 minute to 
completely dry and then to the coverslipper, eliminating, again, ethanol and 
xylene. Both these two steps (and those dealing with tissue processing) have 
made my laboratory (and several others) absolutely xylene (or xylene 
substitutes) free.4- I have always preferred film over glass for speed and 
simplicity. No problems with "coverslip peeling" if the amount of delivered 
xylene (the only instance when I use xylene) is correct.5- Finally, never ask 
for input to a "vendor" for they all have "tainted" opinions at different 
levels and will systematically recommend the products they sell. Do your 
homewotk and ask users, not sllers.I also hope this will helpRené 

On Thursday, March 14, 2019 3:59 PM, Gudrun Lang via Histonet 
 wrote:
 

 Hi Terri,
Have you actually used the Spectra from Leica? This is the follower of
ST5020 and CV5020.
I am also interested in some feedback on this instrument. 

Gudrun Lang

-Ursprüngliche Nachricht-
Von: Terri Braud via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Gesendet: Donnerstag, 14. März 2019 20:10
An: 'histonet@lists.utsouthwestern.edu'
Betreff: [Histonet] Stainer vs. Stainer

Hi Alison - 
I've used both stainers and like both of them a lot.  Both were super
reliable and easy to use.  However, coverslipping is a different story.
I've used both film and glass.  About film - super quick, super easy, but -
the purity of the xylene used to coverslip from film must be absolute.
Anyone who has experienced film pulling off the slides in storage had a
miniscule portion of water carried down the acohols and into the xylene. If
it were glass, the process is a bit more forgiving of water contaminent. The
absolute alcohols leading to the end xylenes must be kept very fresh.  I
kept film slides for over 20 years, no problem.  If you are looking into
digital pathology, I would check with vendors to see if film is acceptable.
I don't know.
As to coverslippers, we've been using the Sakura glass now for 10 years and
love it.  I can't compare it to the newer Leica Glass, but 10 years ago my
techs all preferred the Sakura because it had fewer moving parts and the
maintenance was easier.  I hope this helps.  Good Luck, Terri

Terri L. Braud, HT(ASCP)
Anatomic Pathology Supervisor
Laboratory
Holy Redeemer Hospital
1648 Huntingdon Pike
Meadowbrook, PA 19046
ph: 215-938-3689
fax: 215-938-3874
Care, Comfort, and Heal

  6. stainer v. stainer (Perl , Alison)
  
Message: 6
Date: Wed, 13 Mar 2019 20:08:14 +
From: "Perl , Alison" 
To: "'histonet@lists.utsouthwestern.edu'"
    
Subject: [Histonet] stainer v. stainer


Hi all
We are getting ready to purchase a new H stainer/coverslipper, and are
considering the Sakura Prisma Plus (tape) and the Leica Spectra (glass).
Does anyone have good or bad feedback on either instrument, and/or tape v.
glass? We've always had glass, but of course the coverslippers need more
maintenance, take longer to dry, more expensive than tape, etc etc. So we
are very interested in tape, but still a little hesitant about the old
problems of yellowing and peeling after 10+ years. Since we're in NY, we
have to keep all slides for 20 years

Any thoughts are appreciated!

Alison Perl, HTL(ASCP)CM
Anatomic Pathology Manager
CareMount Medical
110 South Bedford Rd
Mount Kisco, NY 10549
(914) 302-8424
ap...@cmmedical.com
www.caremountmedical.com



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Re: [Histonet] ER/PR question

[Rene J Buesa via Histonet]

Time alone is not the only cause for antigen decay for it is also associated 
with storage conditions and, as such, you cannot "predict" % decay based on 
time alone.Just try one block and let the pathologist decide based on the 
usefulness of what he sees in that try test.René 

On Thursday, February 7, 2019 8:23 AM, "Heckford, Karen - SMMC-SF via 
Histonet"  wrote:
 

 Good Morning,
One of my Pathologists wants me to do some ER/PR's on 9 year old tissue blocks. 
 I know ER and PR can be sensitive are the IHC's going to work or will there be 
too much antigen decay?

Thanks,

Karen Heckford HT ASCP CE
Lead Histology Technician
St. Mary's Medical Center
450 Stanyan St.
San Francisco, Ca. 94117
415-668-1000 ext. 6167
karen.heckf...@dignityhealth.org

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Re: [Histonet] test

[Rene J Buesa via Histonet]

If you add anything, and I mean ANITHING, to your message, the message will not 
be posted.I recommend you to post your problem again without the 
screenshot.René 

On Friday, November 16, 2018 11:27 AM, "Campbell, Tasha M. via Histonet" 
 wrote:
 

 Thanks.  So I got a copy of this email I sent.  But I sent my question twice 
and did not get the copy.  I put a screen shot in the message.  Could that mess 
it up?

 
 
 
Tasha Campbell, B.S.,HTL(ASCP)
Frederick Gastroenterology Associates
310 W. 9th St.
Frederick, MD 21701
301-695-6800 ext. 144 
-Original Message-
From: Eileen Akemi Allison [mailto:akemiat3...@gmail.com] 
Sent: Friday, November 16, 2018 11:01 AM
To: Campbell, Tasha M.; Histonet
Subject: Re: [Histonet] test

** CAUTION: This email originated from outside of Frederick Memorial Hospital. 
DO NOT click on links or open attachments if you do not recognize the sender. **


I received it.  I have had the same problem lately.  I didn't get a copy of the 
email sent.  It would be nice to know that it was received. Think they changed 
their format.

Akemi Allison

> On Nov 16, 2018, at 7:25 AM, Campbell, Tasha M. via Histonet 
>  wrote:
>
> I keep trying to send an email for help with an issue I am having and its not 
> going through. It says the email has been sent but I am not getting the email 
> and no one has responded.  I am testing to see if this message goes through.
>
>
>
>
> Tasha Campbell, B.S.,HTL(ASCP)
> Frederick Gastroenterology Associates
> 310 W. 9th St.
> Frederick, MD 21701
> 301-695-6800 ext. 144
>
>
> CONFIDENTIALITY NOTICE: This electronic mail transmission and any 
> accompanying data files is confidential and is intended exclusively for the 
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> information that is proprietary, privileged or confidential or otherwise 
> legally exempt from disclosure. If you are not the named addressee or you 
> otherwise have received this message in error, you are not authorized to 
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> and delete all copies of this message. Receipt by anyone other than the named 
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Re: [Histonet] Cost per test averages

[Rene J Buesa via Histonet]

 Terri:
All you wrote is absolutely true BUT has nothing to do with the "negotiating" 
aspect Charlie us asking about.He asked about the price he wants to pay and 
that will depend on his purchase volume that could determine, if his' is large 
enough, may entice the seller to reduce his profit margin.
René
On Monday, November 12, 2018, 9:57:37 AM EST, Terri Braud via Histonet 
 wrote:  
 
 I find it very peculiar to be lectured by a Roche representative on including 
reagent management when calculating stain costs, considering Roche's yearlong 
ongoing issues with reagent supply problems and massive recalls.  I've yet to 
have one month with no reagent supply problems from Roche/Ventana.  Failed 
dispensers, multiple recalls, major delays in deliveries of supplies (the most 
recent was a TWO MONTH BACKORDER on a routinely run antibody) and the list goes 
on.
Roche needs to get their own house in order before they come on a technical 
list-serve to lecture.
Just saying...

Terri L. Braud, HT(ASCP)
Anatomic Pathology Supervisor
Laboratory
Holy Redeemer Hospital
1648 Huntingdon Pike
Meadowbrook, PA 19046
ph: 215-938-3689
fax: 215-938-3874
Care, Comfort, and Heal

Today's Topics:
Message: 2
Date: Sat, 10 Nov 2018 14:54:19 -0500
From: "Frazier, John" 
To: Charles Riley 
I?m a vendor that sells an H staining product, so I?m not going to attempt to 
give you what the average cost per of a H stained slide.
What I will tell you is that when looking at cost per slide you need to 
calculate more than your consumables. Those are going to be your capital cost. 
You also need to calculate in your labor cost. Those are gonna be your 
operational dollars.
The reason why I say that is that some strainers are more efficient than other 
stainer. Not just in the staining process itself but in the overall 
maintenance, reagent management and waste control cost.
Remember when using labor dollars you want to calculate that using fully 
burdens dollars.
Be complete in the way that you calculate your overall cost per slide

John Frazier, MBA, MT(ASCP)
Strategic Workflow Consulting
Roche Diagnostics



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Re: [Histonet] REPEATS IN IHC

[Rene J Buesa via Histonet]

Ginny:It seems you are asking for a "problem" average and that will vary per 
laboratory and how the whole process is carried out.Section washed out from the 
slide usually is consequence of poor processing or sectioning and has nothing 
to do with IHC, although, perhaps, a wrongly performed HIER can cause it.Wrong 
block is a mistake also having nothing to do with IHC and is always caused by 
an inattentive HT not paying attention to the process.My point is that, 
essentially, the lab should try to get to "0" mistakes and any one has to be 
documented in the QC record but it will be interesting to find out the 
"mistakes rate" you find and I think you should share it with all.René 

On Friday, October 26, 2018 4:41 PM, Kurth Virginia L. via Histonet 
 wrote:
 

 Hello, I was wondering if anyone in the histology laboratory (IHC) has a limit 
on how many times one would repeat a stain; for example but not
Limited too:  The tissue fell off, negative control, wrong block,etc.. for IHC 
DAB and/or ISH.  Thanks Ginny
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Re: [Histonet] Bouin's Fixative Substitute

[Rene J Buesa via Histonet]

If "New Bouin" has no pricric acid it is NOT Bouin. You can call it whatever 
you want but it is not Bouin nor it can have the "quality" provided by picric 
acid without picric acid.What you wrote is simply a deceiving"sales pitch".René
 

On Wednesday, October 24, 2018 6:10 PM, Linda Miller via Histonet 
 wrote:
 

 New! Bouin's Fixative Substitute is a low-hazard replacement for Bouin's
Fixative.  This substitute provides all the benefits of Bouin's without the
use of picric acid. Available in a variety of sizes and pre-filled specimen
containers.

Available at IMEB, Inc.


-Original Message-
From: histonet-requ...@lists.utsouthwestern.edu
[mailto:histonet-requ...@lists.utsouthwestern.edu] 
Sent: Friday, October 19, 2018 10:00 AM
To: histonet@lists.utsouthwestern.edu
Subject: Histonet Digest, Vol 179, Issue 16

Send Histonet mailing list submissions to
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When replying, please edit your Subject line so it is more specific than
"Re: Contents of Histonet digest..."


Today's Topics:

  1. toluene substitute (Michelle Jamison)
  2. RUO antibody (Erin McCarthy)
  3. Re: Restaining old Heamatology smears (Eddie Martin)
  4. Re: BMP-2 protocol for bone (Eddie Martin)


--

Message: 1
Date: Thu, 18 Oct 2018 14:48:50 -0600
From: Michelle Jamison 
To: "histonet@lists.utsouthwestern.edu"
    
Subject: [Histonet] toluene substitute
Message-ID: 
Content-Type: text/plain; charset=utf-8; format=flowed

Hi All, I am from industry. We are interested in developing an instrument
that stains then prepares microscope slides to be coverslipped. If you do
coverslip, what substitute do you use instead of toluene to go to mounting
media? Thank you! Michelle
-- 

All the best to you,

Michelle Jamison

/Bio Research Associate /

/ELITechGroup Inc. / ???370 W. 1700 S.?? ???Logan Utah ??? USA

m.jami...@elitechgroup.com  ? 
_www.elitechgroup.com  _

Tel : +1.435.752.6011 Ext: 1474

Logo


--

Message: 2
Date: Thu, 18 Oct 2018 16:30:14 -0500
From: Erin McCarthy 
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] RUO antibody
Message-ID:
    
Content-Type: text/plain; charset="UTF-8"

Hi All,

I am looking to see if anyone has had any experience with the antibody AXL.
We are trying to work it up on our Roche Discovery Ultra and it does not
seem like it stains both nuclear and cytoplasic cell features. We only seem
to see cytoplasmic staining, and in general most tumors are showing negative
results.

I currently am using the Ultraview kit for optimization, our pathologist
likes CC2 for 44 min, Ab. Inc for 40 min using a 1:100 dilution.

Anyone have any suggestions that might help? If you want more detailed info
I can provide as well!

-- 

Erin McCarthy, HT (ASCP)
Pathology Lab Supervisor

Tempus Labs
600 W. Chicago Ave.
Chicago IL 60654
Ph:(312) 638-6344 Ext.3835

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Message: 3
Date: Thu, 18 Oct 2018 18:20:53 -0400
From: Eddie Martin 
To: "Reilly, Laurie" 
Cc: "Histonet@lists.utsouthwestern.edu"
    
Subject: Re: [Histonet] Restaining old Heamatology smears
Message-ID: <0e7f124e-9821-4a32-a438-63f45ad45...@gmail.com>
Content-Type: text/plain;    charset=utf-8

Hi Laurie,

Blood is considered a tissue so it is very much still Histo related. 

Regarding your smears, this also occurs in our hematopathology laboratory
from time to time when we are scanning old slides and there?s a bubble under
the coverslip that won?t let us take images with our digital scope. 

It may very well be that there is still a resinous permount film on your
slides from the old coverslip. This may simply be removed by leaving the
smear in xylene some additional time to remove the 

Re: [Histonet] Cost calculations

[Rene J Buesa via Histonet]

Charles:There is the "easy way" and the "hard way" and the differences are 
minuscule:1- add what cost you ALL the ingredients for each aspect you want to 
determine2- add the salary of the people doing the task related to productivity 
(blocks or slides or whatever per hour)3- add-up 1 + 24- divide by the number 
of "units" produced.Under separate cover I am sending an article I wrote on the 
subject
René 

On Tuesday, October 23, 2018 6:26 PM, Charles Riley via Histonet 
 wrote:
 

 How does everyone calculate their cost to create things in your lab?

I was tasked to figure out how much it costs us to make the following
1. A paraffin block
2. A single H slide
3. Average cost of an IHC slide
4. Average cost of a Special stain slide

What I need to figure out is beside the cost on the inventory to create
these items what other factors do I need/should use to calculate final
costs (i.e. tech time, energy costs, machine maintenance)?

-- 

Charles Riley BS  HT, HTL(ASCP)CM

Histopathology Coordinator/ Mohs
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Re: [Histonet] VIP issue

[Rene J Buesa via Histonet]

Gudrun:
I have used/recommended for 10 years now graded isopropyl (2-propanol) alcohol 
to dehydrate and before paraffin ONLY and when I went to Australia in 2011 
found out that they had been using the same protocol for years.
You just grade 2-propanol in the same way you grade ethanol (60%→80%→90%→100%) 
and continue DIRECTLY to paraffin.
Usually I dedicate one "hot" (paraffin) VIP station with a 1:1 of 2-propanol + 
paraffin mixture to facilitate the infiltration or (better yet) one "hot" 
station with a 1:1 mixture of 2-propanol + pure mineral oil (= paraffin of low 
molecular weight) and the infiltration is really excellent.
In this way you will be able to eliminate BOTH xylene and any other "friendly" 
clearing agent (ALL are toxic lat different evels). 2-propanol is NOT toxic AT 
ALL!
I have ALSO stopped using xylene to de-wax sections before staining and 
substituted it with a 2% aq. sol. of dishwashing soap and go to water → 
staining after (I can send you the detailed protocol / publications).
AFTER staining I just rinse the stained slides in 2-propalo → shake it well → 
to dry oven at 60ºC for 10 minutes → coverslip.
My lab was TOTALLY xylene free, and you can do the same thing. If you are 
hesitant just TRY these methods with only few blocks / sections / stained 
sections and convince yourself.
Best regards
René 

On Friday, October 12, 2018 1:13 PM, Gudrun Lang via Histonet 
 wrote:
 

 Thank you for all the good hints and advices.

It has to be a reagens-issue, because the second instrument was also
concerned. 
But we had no similar problems for decades, although we use 96% as
cleaning-ethanol after ShellSol. 
We change the reagenses after 5 runs, once a week. 

We soon are forced to end using ShellSol, because the company stops the
production. We don't like to switch to xylene. 
Which xylen-substitutes are recommended for the VIP besides of Tissue-Clear
(Sakura)?

Best wishes
Gudrun 


-Ursprüngliche Nachricht-
Von: Gudrun Lang via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Gesendet: Donnerstag, 11. Oktober 2018 18:58
An: histonet@lists.utsouthwestern.edu
Betreff: [Histonet] VIP issue

Dear all!

I have a question for those, who are familiar with the VIPs from Sakura. 

Last time we changed the reagenses the cleaning-ethanol (96%) was very milky
and even was full of many small particles. It was a paraffin-soup.

What is the cause for such a case?  We have been using the organic solvent
(ShellSol) for decades as xylensubstitute. 

Maybe the quality has suffered and the ability of solving paraffin has
decreased. But are there other explanations? Maybe a malfunction of the
instrument?

 

Thanks in advance

Gudrun Lang

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Re: [Histonet] H for H. pylori?

[Rene J Buesa via Histonet]

Paula:"Optimizing" H to detect H. piloris is a rout you should not attempt 
because H cannot be "optimized" for that.Giemsa is one way or, even better, 
modified Steiner stain. The only disadvantage modified Steiner has is using 
radioactive uranium nitrate, but I developed a procedure where 0.02% aq. 
phosphotungstic acid solution can be used instead. IHC will be way more 
expensive.
René 

On Tuesday, October 9, 2018 2:03 PM, P Sicurello via Histonet 
 wrote:
 

 Good Morning Listers,

Has anyone out there optimized an H for H. pylori?

Sure we can run a giemsa or and IHC but what fun is that?

Send me your ideas and make the day of our GI pathologists.

Thanks oodles.

Sincerely,

Paula Sicurello, HTL (ASCP)CM

Histotechnology Specialist

UC San Diego Health

9300 Campus Point Drive

La Jolla, CA 92037
(P): 858-249-5610



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Re: [Histonet] FW: Leica Bond Max artifact

[Rene J Buesa via Histonet]

Dear Dr. Rosen:
I see nothing wrong in your protocol but your photo 0950 = 09501 seems as if 
there are 2 superimposed sections, one with a lot of brown particles? Is this 
artifact you are referring to? Seems "too organized" to be an artifact but it 
is brown and not red.
These "non DAB" reagents sometimes develop a sort of precipitate even within 
their expiration date. Why have you decided to use this red reagent instead of 
DAB? That could be a better option.
Other than that I am also completely baffled by your problem.
René


 

On Tuesday, October 9, 2018 1:15 PM, Linda Margraf via Histonet 
 wrote:
 

 Here is a message I am posting for Dr. Rosen:
Hello,

I am a longtime admirer of this list, and I am sure someone on here can help us 
out

We have an artifact on our immunohistochemistry (IHC) slides.

I will attempt to attach pictures using the upload tool. In case that doesn’t 
work, I am attaching a google drive link to some jpgs. The artifact is usually 
golden brown to red/pink in color, is clustered, appears both on top of the 
tissue and around it. If we do an aggressive rundown, it appears less red/brown 
and bluer.

We are using a Leica Bond MAX IHC system which is about a year old. We usually 
do two runs a day and 95% of the time we only use a red detection kit. We only 
see this on our red detection kit on our Leica bond. The brown is unaffected. 
The corresponding H slides are fine.
We started having this artifact after a month or two of production. 85% of the 
time we have some sort of artifact. Sometimes its perfect, sometimes it makes 
the tissue unreadable. We haven’t figured out any precipitating factors.

We have tried a number of things, but we still can’t get control of this.
Here is a brief overview of our protocols:
- Processor: Leica ASP300s tissue processor.
-Parrafin = Mercedes HWWX (dual purpose embedding and infiltration parrafin) 
melting point 57c – we tried switching, and it didn’t help.
- We only use distilled water in our baths
- Mercedes Starfrost charged slides
- Oven for 30 min at 62c
- We always use a cold red detection kit
-Sometimes our runs are delayed but not more than 4 hours
- Rundown is done in our stainer (standardized), and slides are coverslipped 
with tape.
- The bulk fluid containers in the bond are cleaned weekly and refrigerated 
between runs.
- The probe is cleaned every 300 slides.
- Mixing wells are cleaned out every two weeks and replaced often.
-Covertiles are well tended to.

We have tried to correct this problem, but we can't seem to fix it.
Any ideas?

https://drive.google.com/open?id=1wUttTweP7SYLRM5pkGOBr0MBzCMrtJSA


Jason R. Rosen, D.O.
Dermatopathologist
Premier Dermatology Partners
4675 Linton Boulevard, Suite 203
Delray Beach, FL 33445
(p) 561-499-5341
(f) 561-499-5343
(e) jro...@totalderms.com


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Re: [Histonet] Pregnancy guide for working in histology

[Rene J Buesa via Histonet]

Both formalin and xylene (and any other dangerous fumes) have to be avoided 
during pregnancy BUT if you have an efficient fumes hood to do grossing, then 
you should monitor exposure. Somebody NOT pregnant should gross with a personal 
formalin badge and, depending on the exposure result, then you may allow the 
pregnant employee to do grossing or not.René 

On Thursday, September 6, 2018 1:08 PM, Carol Torrence via Histonet 
 wrote:
 

 Good morning!

Could some of you chime in on guidelines you go by for those employee that are 
expecting a baby.  I have removed employee from xylene exposure during staining 
and cover slipping but am on the fence about grossing.  At this time the 
employee has been removed from grossing.  All grossing is done under an exhaust 
hood.  Our exposure badges have always read well below limits.  Thanks in 
advance!

Carol M. Torrence, HT(ASCP)

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Re: [Histonet] Unstained slides

[Rene J Buesa via Histonet]

On this issue of lost of antigenicity, never forget air oxygen!René 

On Monday, September 3, 2018 11:11 AM, "Frazier, John" 
 wrote:
 

 Interesting that you stated that, I was at the university of Colorado
this past week and was speaking with the medical director of the
pathology department. We actually started talking about unstained
slides and their storage conditions. We actually spoke of the histonet
discussions around unstained slide storage.  He stated to me that due
to the elevation and lack of humidity in Denver that the antigenicity
of unstained slides has been up to multiple years. This is due to, as
you stated, water in the tissue.

Sent from my iPad

> On Sep 3, 2018, at 9:42 AM, Cartun, Richard  
> wrote:
>
> It appears that the presence of water, both endogenously and exogenously, 
> plays a central role in the loss of antigenicity in stored unstained slides 
> (see reference below).  Labs that are experiencing significant loss of 
> immunoreactivity in their unstained slides should check their tissue 
> processing.
>
> Xie R, Chung J-Y, Ylaya K, et al.:  Factors influencing the degradation of 
> archival formalin-fixed, paraffin-embedded tissue sections.  J of Histochem 
> Cytochem 2011; 59:356-365.
>
> Richard
>
> Richard W. Cartun, MS, PhD
> Director, Histology & The Martin M. Berman, MD Immunopathology & Morphologic 
> Proteomics Laboratory
> Director, Biospecimen Collection Programs
> Assistant Director, Anatomic Pathology
> Hartford Hospital
> 80 Seymour Street
> Hartford, CT  06102
> (860) 972-1596
> (860) 545-2204 Fax
>
> -Original Message-
> From: Rene J Buesa via Histonet [mailto:histonet@lists.utsouthwestern.edu]
> Sent: Sunday, August 19, 2018 2:09 PM
> To: Frazier, John; Terri Braud
> Cc: histonet@lists.utsouthwestern.edu
> Subject: Re: [Histonet] Unstained slides
>
> This is an email from Outside HHC. USE CAUTION opening attachments or links 
> from unknown senders.
>
> Everything has been pointed out is correct BUT also pivot on "how the 
> unstained slides are kept".Kept in a box their "useful life" is quite short 
> (not beyond 1 week at the most).Kept at -80ºC I have used them after years of 
> being stored the principle being of deep-freezing and this is "standard 
> procedure" for IDF "+ controls".Kept in a Coplin jar filled with mineral oil 
> or paraffin covered I have used them after months of being stored the 
> principle being that, isolated from air oxygen, epitopes do not oxidize 
> ("weaken") of if they do, the rate is greatly slowed.On the other hand, 
> usually, unstained slides are kept for only few days in the event that, lets 
> say within a week, the PT decides to order some special procedure and 
> sometimes it is impossible "return" to the original block many times "almost 
> exhausted".Properly done storing unstained slides are extremely useful.René
>
>    On Sunday, August 19, 2018 1:52 PM, "Frazier, John via Histonet" 
> wrote:
>
>
> I agree with Tim as well. This is what we advise our clients to do. It takes 
> some coordination with the pathologist, but it is the best strategy for 
> reducing unnecessary unstained slides. In the studies that we have performed 
> only 10% of the unstained slides that are cut are you and the 90% are are it 
> takes some coordination with the pathologist, but it is the best strategy for 
> reducing unnecessary unstained slides. In the studies that we have performed 
> only 10% of the unstained slides that are cut are you and 90% are thrown away 
> thrown away.
> Several laboratories that I have visited in order to reduce the amount of 
> wasted tissue when refacing the blocks, is to reseal the blocks with liquid 
> paraffin, that have scant or small amounts of tissue in the block, such as 
> the needle core biopsy.
> Bottom line on this issue is to educate the pathologist, and not water and 
> stain slides except in rare occasions
>
> Sent from my iPhone
>
>> On Aug 17, 2018, at 14:07, Terri Braud  wrote:
>>
>> I'm with Tim Morken on this one. The variability of antigenicity in storage 
>> is so wide open, and there really is no recent data, so we just make a point 
>> of educating our techs on not wasting tissue/levels during sectioning.  If 
>> the techs feel that the residual tissue in the block is in danger of being 
>> exhausted, we communicate with our pathologists on how best to handle any 
>> requests.  Unstained slides was time, money, and storage and we are better 
>> off without them.
>>
>> Terri L. Braud, HT(ASCP)
>> Anatomic Pathology Supervisor
>> Laboratory
>> Holy Redeemer Hospital
>> 1648 Huntingd

Re: [Histonet] IHC Stainer

[Rene J Buesa via Histonet]

Get a DAKO IHC stainer (even if it is "refurbished")René 

On Thursday, August 30, 2018 9:56 AM, Sandra Cheasty via Histonet 
 wrote:
 

 Hello all,
                We are in the market for a new IHC stainer. (Our Lab Vision 720 
is no longer reliable.) We average about 20 slides a day, but sometimes get 
research projects that are 50+ slides.  Not interested in a closed system; it 
is for a veterinary histology lab, and we need the flexibility to tweak 
antibodies for dogs, cats, horses, and zoo animals. I'd really appreciate 
feedback from users who are in a similar lab setting.

Cheers!
Sandy

Sandra J. Cheasty, HT (ASCP)
Histology & Necropsy Supervisor
UW-Madison, School of Veterinary Medicine

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Re: [Histonet] Unstained slides

[Rene J Buesa via Histonet]

Everything has been pointed out is correct BUT also pivot on "how the unstained 
slides are kept".Kept in a box their "useful life" is quite short (not beyond 1 
week at the most).Kept at -80ºC I have used them after years of being stored 
the principle being of deep-freezing and this is "standard procedure" for IDF 
"+ controls".Kept in a Coplin jar filled with mineral oil or paraffin covered I 
have used them after months of being stored the principle being that, isolated 
from air oxygen, epitopes do not oxidize ("weaken") of if they do, the rate is 
greatly slowed.On the other hand, usually, unstained slides are kept for only 
few days in the event that, lets say within a week, the PT decides to order 
some special procedure and sometimes it is impossible "return" to the original 
block many times "almost exhausted".Properly done storing unstained slides are 
extremely useful.René 

On Sunday, August 19, 2018 1:52 PM, "Frazier, John via Histonet" 
 wrote:
 

 I agree with Tim as well. This is what we advise our clients to do. It
takes some coordination with the pathologist, but it is the best
strategy for reducing unnecessary unstained slides. In the studies
that we have performed only 10% of the unstained slides that are cut
are you and the 90% are are it takes some coordination with the
pathologist, but it is the best strategy for reducing unnecessary
unstained slides. In the studies that we have performed only 10% of
the unstained slides that are cut are you and 90% are thrown away
thrown away.
Several laboratories that I have visited in order to reduce the amount
of wasted tissue when refacing the blocks, is to reseal the blocks
with liquid paraffin, that have scant or small amounts of tissue in
the block, such as the needle core biopsy.
Bottom line on this issue is to educate the pathologist, and not water
and stain slides except in rare occasions

Sent from my iPhone

> On Aug 17, 2018, at 14:07, Terri Braud  wrote:
>
> I'm with Tim Morken on this one. The variability of antigenicity in storage 
> is so wide open, and there really is no recent data, so we just make a point 
> of educating our techs on not wasting tissue/levels during sectioning.  If 
> the techs feel that the residual tissue in the block is in danger of being 
> exhausted, we communicate with our pathologists on how best to handle any 
> requests.  Unstained slides was time, money, and storage and we are better 
> off without them.
>
> Terri L. Braud, HT(ASCP)
> Anatomic Pathology Supervisor
> Laboratory
> Holy Redeemer Hospital
> 1648 Huntingdon Pike
> Meadowbrook, PA 19046
> ph: 215-938-3689
> fax: 215-938-3874
> Care, Comfort, and Heal
>
> Today's Topics:
>  7. Re: Unstained slides - how long are they good for?
>      (Morken, Timothy)
>
> Message: 7
> Date: Fri, 17 Aug 2018 15:16:00 +
> From: "Morken, Timothy" 
> To: P Sicurello 
> Subject: Re: [Histonet] Unstained slides - how long are they good for?
>
>
> Paula, since it is variable we strive to not have unstained slides. We had 
> kept them indefinitely, then when storage was overwhelming us we reduced it 
> to 2 months maximum. Now we require request for unstained to be ordered in 
> the system and delivered to the pathologist. We do not hold any in the lab. 
> We recut when new stains are ordered. In the past we had routinely cut extras 
> "just in case" but ended up with thousands of unstained slides that were 
> never used. Instead we trained everyone to reduce wastage and get good 
> sections from a cut block with minimal facing. We have not stored unstained 
> sections for many years and they do not seem to be missed.
>
> Tim Morken
> Pathology Site Manager, Parnassus
> Supervisor, Electron Microscopy/Neuromuscular Special Studies
> Department of Pathology
> UC San Francisco Medical Center
>
>
> -Original Message-
> From: P Sicurello via Histonet [mailto:histonet@lists.utsouthwestern.edu]
> Sent: Thursday, August 16, 2018 4:49 PM
> To: HistoNet
> Subject: [Histonet] Unstained slides - how long are they good for?
>
> Hello My Fellow Histologists,
>
> Happy Friday Eve.
>
> The question has come up..  How long are *unstained* slides good for?
> Not for H but tests like IHC and molecular testing.  These slides have
> been cut, stored at room temperature, not sealed in anyway, and kept in a
> cardboard box.
>
> Please let me know what your opinions are and what your retention policy is
> concerning *unstained* slides.
>
> Thanks oodles.
>
> Sincerely,
>
> Paula Sicurello, HTL (ASCP)CM
>
> Histotechnology Specialist
>
> UC San Diego Health
>
> 200 Arbor Drive
>
> San Diego, CA 92103
>
> (P): 619-543-2872
>
>
>
> *Confidentiality Notice*: The information transmitted in this e-mail is
> intended only for the person or entity to which it is addressed and may
> contain confidential and/or privileged material.  Any review,
> retransmission, dissemination or other use of or taking of any action in
> reliance upon this information by persons or entities other 

Re: [Histonet] paraffin temperatures

[Rene J Buesa via Histonet]

The processor screen temperature reflects the actual reading of an embedded 
electronic temperature probe and, as such, will be more accurate (always at the 
same place, in the same way, with constant precision) something rarely obtained 
manually. Therefore, use the temperature reading from the screen.René 

On Saturday, August 18, 2018 12:55 PM, Carol Kowalcyk via Histonet 
 wrote:
 

 When recording paraffin temperatures...do you record all 4 temps. from the 
processor screen and/ or take the temps by hand in the 4 paraffin baths and 
record or do you take the temp. of paraffin actually in the retort before 
draining?
Thanks!
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Re: [Histonet] Trouble shooting question

[Rene J Buesa via Histonet]

 Check fixation time and specially IF the Bx was left to dry before 
fixation.René

On ‎Thursday‎, ‎July‎ ‎12‎, ‎2018‎ ‎12‎:‎38‎:‎28‎ ‎PM, Charles Riley via 
Histonet  wrote:  
 
 My pathologists says that 1 out of 100 (his estimate) of our FNA's
cellblocks  apears to be "cooked" when looking at the slides.  The process
has never been changed and all reagents are the same all the time.

 What is/are the possible cause(s) and what can be done to prevent this 1
out of 100 situation?




-- 

Charles Riley BS  HT, HTL(ASCP)CM

Histopathology Coordinator/ Mohs
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Re: [Histonet] Disposing Paraffin

[Rene J Buesa via Histonet]

 We incinerated itRené

On ‎Friday‎, ‎June‎ ‎29‎, ‎2018‎ ‎03‎:‎49‎:‎50‎ ‎PM, Paula via Histonet 
 wrote:  
 
 Hello all.this is an on-going topic with our lab. 

 

If anyone can comment about your experience with paraffin waste, I
appreciate that in advance. 

 

Does your lab or state requirements classify paraffin waste as "hazardous,
bio-hazardous, RCRA or other"?  Do you dispose as part of Red Bag trash for
autoclave or tissue pathology waste or other hazardous waste for
incineration? 

 

Paula

Lab Manager for Biopath Medical Group in Fountain Valley CA

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Re: [Histonet] Microscope objectives

[Rene J Buesa via Histonet]

Not all objectives "are created equally"."In the beginning" (as in Genesis) the 
objective "was created" to be used on microscopes with essentially two 
different focal distances, i.e. 160mm and 170mm. By the 1950's the ∞ corrected 
objective appeared to simplify somewhat the situation BUT every manufacturer 
tried that all their objectives were "par-focal" meaning that you could switch 
from one objective to the next and the virtual image remained "in focus".That 
is the crux of the issue: a virtual image permanently in focus, and that cannot 
be obtained between objectives from different manufacturers because their sizes 
(lengths) are different, and their focal points may also differ.So, putting it 
simply, unless you are willing to refocus with either your "macro" or "micro" 
knob (focusing wheel) you will need to buy all your objectives from the same 
manufacturer (brand).René 

On Tuesday, June 19, 2018 8:58 AM, Charles Riley via Histonet 
 wrote:
 

 Are all microscope objectives universal between brands or do you have to
buy the objectives from the company that manufactured it?

-- 

Charles Riley BS  HT, HTL(ASCP)CM

Histopathology Coordinator/ Mohs
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Re: [Histonet] Radiation sterilization and IHC

[Rene J Buesa via Histonet]

 I believe that is an extreme and unnecessary step. Formalin itself is an 
adequate sterilizer, the only problem would be with "prions" and 
Creutzfeldy-Jakob disease cases are not only extremely rare, but always known 
or suspected to histotechs. René. 

On ‎Monday‎, ‎April‎ ‎23‎, ‎2018‎ ‎11‎:‎56‎:‎39‎ ‎AM, Mike Tighe via 
Histonet  wrote:  
 
 It was recently suggested to me that we might try to use our irradiator to 
sterilize (not fix) tissues rather than Formalin solution. Obviously we would 
need to validate this but has anyone tried this prior to IHC or IF?

Thanks!
Mike
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Re: [Histonet] Congo Red

[Rene J Buesa via Histonet]

You will need a polarizing filter. The size is just the necessary to cover your 
light source. René 

On Friday, March 23, 2018 10:27 AM, Frances Pearsall via Histonet 
 wrote:
 

 Can anyone tell me what kind (or size)  glass filter to use to perform 
Puchtlers Congo Red? And are the filters re-useable?

Thank you,
Fran Pearsall
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Re: [Histonet] Wright-Giemsa for sections?

[Rene J Buesa via Histonet]

Tyrone:Under separate cover I am sending a procedure I published about this 
method.René 

On Friday, February 2, 2018 4:44 PM, Tyrone Genade via Histonet 
 wrote:
 

 Hello,

Can the Wright-Giemsa stain be used on fixed, paraffin embedded sections?
Does anyone have a protocol?

I want to examine hematopoietic tissue of fish, i.e. the head kidney. No
smears or imprint possible. I would like to use Wrights so I can use the
same stain for blood smears.

Thanks

-- 
Tyrone Genade
Orange City, Iowa
tel: (+1) 712 230 4101
http://tgenade.freeshell.org

Romans 6:23: The gift of God is eternal life through Christ Jesus our Lord.
To find out how to receive this FREE gift visit http://www.alpha.org.

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Re: [Histonet] Negative IHC Control for Her2

[Rene J Buesa via Histonet]

You are right but, your point is?Regardless, the pathologist is the responsible 
for his/her diagnosis/interpretation and the liability is his/hers.Remember 
that for us histotechs, our "client" is the pathologist (always remembering the 
patient behind the whole process) and our essential duty is to provide the 
pathologist what s/he needs to being able to make the best and more accurate 
diagnosis.Follow his/her instructions and your life will be much simpler and 
less stressful.René 

On Tuesday, November 21, 2017 4:21 PM, "Lindrud, Scott via Histonet" 
 wrote:
 

 Hi All,

We are having an internal debate regarding Her2 IHC control tissue in our lab.  
We run a MTA (multi tissue array) consisting of 0+, 1+, 2+, and 3+ Her2 
staining tissue taken from lumpectomy/resection cases.  I'm in the process of 
searching for more tissue to use in future control blocks and it can be 
difficult to find tissue that is 0+ and 3+.

I've discussed this with our pathologist in charge of Histology and he says 
that we don't need to run all 4 of the cores as controls.  He says all we need 
to run is a positive control and a negative control.  He says the positive 
control could be a 2+ or 3+ and the negative control could be either a 0+ OR a 
1+.

I respectfully disagreed with him and said the negative control is not meant 
for accessing staining interpretation but to verify the sensitivity and 
specificity of the actual antibody-antigen reaction.  I said a 1+ is still 
staining the tissue where there is no staining in a 0+ reaction.  The 1+ is not 
a negative staining reaction, but a negative interpretation.  The pathologist 
says I'm wrong.

The CAP in its checklist says "It is also important to assess the specificity 
of each antibody by a negative tissue control, which must show no staining of 
tissues known to lack the antigen".    To me, a 1+ Her2 staining reaction shows 
that that tissue has antigen and should not be used as a negative control.

So, after saying all that, can/should  1+ Her2 breast tissue be used as a 
negative tissue control?  Seems pretty straight forward to me, but I'm just a 
Cytotechnologist/Histotech.

Thanks for any input!

Scott

Scott A. Lindrud, MLS(ASCP)CM CTCM
Histopathology Technical Specialist/Cytotechnologist
Rice Memorial Hospital
301 Becker Ave SW
Willmar, MN 56201
WP(320)231-4406
Fax(320)231-4861
scott.lind...@rice.willmar.mn.us




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Re: [Histonet] Microtomy question

[Rene J Buesa via Histonet]

It is impossible to say how deep to cut into a block. First the PA must be sure 
s/he was sure the tumor/lesion was in the piece submitted to process. If that 
is the case, the one who casted the block must be sure the lesion is as near 
the block surface as possible in a way that when the block is trimmed, the 
lesion is macroscopically visible.The rest depends on the pathologist is s/he 
can make the diagnosis with the slides first submitted or if deeper sections 
are required.Usually deeper sections are requested when the tumor or abnormal 
tissue is visible and more information is required. It is very rare that a 
pathologist requests deeper to find the lesion, unless trimming has been so 
superficial that the slide barely reveals the processed tissue.Usually lesion 
will be near the center of the tissue processed because the PA leaves tissue 
around the lesion. If this practice is followed, the histotechs should trim 
until the sections closely has the same size (area) as the processed tissue.If 
the pathologist asks for deepers several times, and s/he is sure there should 
be a lesion in the section, keep presenting sections until the lesion is 
found.As an advise, do not discard sections and keep at least 1 every 5 
sections to prevent a very small lesion is lost for ever and the diagnosis 
cannot be made.Consult always with your pathologist, especially asking if there 
should be a lesion, because the whole problem may reside in the fact that the 
PA did not include what the pathologist is looking for in the tissue submitter 
for processing.René 

On Wednesday, July 19, 2017 9:30 AM, Charles Riley via Histonet 
 wrote:
 

 We currently are having issues with our pathologists asking for deeper
levels. They complain that they are not finding tumors that should be there
and need us to go deeper.

Does anyone have any suggestions on how far into a block I should explain
to my techs to go in order to help them get over the fear of going too deep
into a block?



-- 

Charles Riley BS  HT, HTL(ASCP)CM

Histopathology Coordinator/ Mohs
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Re: [Histonet] A Question About Paraffin Times

[Rene J Buesa via Histonet]

I consider taking out → freezing →  melting → casting blocks is worse than 
leaving the tissues in molten paraffin.It seems that your Monday tissue 
processing ends on Saturday. I suggest to process your tissues with a "delay" 
(most tissue processers have this feature) and leave  them more time in 
formalin and make coincide their ending time with your Monday start time.René 

On Tuesday, May 16, 2017 11:51 AM, P Sicurello via Histonet 
 wrote:
 

 Good Morning Listers,

I am asking the collective wisdom of the Histonet this question:


Is it better to remove baskets from the processor on Saturday morning and:

A.  Let the cassettes freeze, then melt them down and embed Monday morning?
OR
B.  Leave the cassettes in molten paraffin and embed Monday morning?


I am of the opinion that leaving the samples (not fatty, like breast cores)
in molten paraffin (62 degrees C)  is bad practice, and causes them to get
"crunchy", among other things.

What do you think?

Thank in advance.

Sincerely,

Paula Sicurello, HTL (ASCP)CM

Histotechnology Specialist

UC San Diego Health

200 Arbor Drive

San Diego, CA 92103

(P): 619-543-2872 <#>



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Re: [Histonet] KOH softener to "decalcify" bone?

[Rene J Buesa via Histonet]

EDTA can be used to decalcify delicate bones, such as bona marrow core 
biopsies.KOH is used to soften keratin (such as in toe nails) but is totally 
ineffective for decalcification.René 

On Monday, May 8, 2017 2:32 PM, Dorothy Hu via Histonet 
 wrote:
 

 I know someone used KOH with EDTA together to decalcify bone.
It is worthwhile to try KOH only to soft bone and kept dynamic labeling.
If anyone had tried successfully, please let us know.
Thanks.


>
> Today's Topics:
>
>    1. Fabric softener to "decalcify" bone? (Angela Lamberth)
>    2. Re: Fabric softener to "decalcify" bone? - Downy has      formic
>      acid?? (Angela Lamberth)
>
>
>
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Re: [Histonet] Cytology pap staining

[Rene J Buesa via Histonet]

Are the smears collected, fixed and treated as "usual" along ALL the steps?René 

On Wednesday, April 19, 2017 12:54 PM, Charles Riley via Histonet 
 wrote:
 

 I have been put in charge of figuring out why our pap stains are light on
the hematoxylin. Everything was filtered and used fresh as per usual using
the same protocol we have used for the past two years.

If anyone has any suggestions as to how to fix this problem I would greatly
appreciate it. If you need more information please let me know and I will
get it for you to help assist you in assisting me

-- 

Charles Riley HT, HTL(ASCP)CM

Histopathology Coordinator/ Mohs
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Re: [Histonet] processor died overnight

[Rene J Buesa via Histonet]

In 100% EthOL the tissues are completely "salvaged" and you can prepare the 
program to continue the steps until melted paraffin.If there are delicate 
tissue perhaps they will be "over-dried" but that is easily "compensated" 
during microtomy.René 

On Wednesday, April 19, 2017 9:01 AM, Lauren Sweeney via Histonet 
 wrote:
 

 Hello Histoworld,

I came in this morning to find that the processor died halfway through process 
last night. The tissues are in 100% ETOH exactly half point. We do have a back- 
up processor. In your professional experiences, would these tissues be 
salvageable? Could I create a new program on the backup processor that finishes 
the process from that point and transfer the tissues over?

Thanks.


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Re: [Histonet] Fw: orders from resident surgeons

[Rene J Buesa via Histonet]

At my hospital we accepted work orders from our pathologists only. If a 
physician resident wanted some test, it has to be approved by our 
pathologist.René 

On Thursday, April 13, 2017 12:18 PM, "Horn, Hazel V via Histonet" 
 wrote:
 

 
I was not clear enough in my question.  I'm not speaking of pathology residents 
ordering tests.  I'm asking if you allow general resident physicians to order 
pathology tests.  Such as gross and micro.

We are a teaching hospital and have never allowed a resident to order pathology 
if they aren't a staff physician (surgeon).

Thanks!



From: Horn, Hazel V
Sent: Thursday, April 13, 2017 8:17 AM
To: histonet
Subject: orders


In the 25+ years I have worked at ACH we have never allowed a resident 
physician to order a pathology test.  I am now being asked why we do this?  I 
have no idea if there is a rule from Medicaid or insurance providers that all 
orders must be from a staff physician.  Does your hospital allow residents to 
place pathology orders?



Hazel Horn

Supervisor of Histology/Autopsy/Transcription

Anatomic Pathology

Arkansas Children's Hospital

1 Children’s Way | Slot 820| Little Rock, AR 72202

501.364.4240 direct | 501.364.1241 fax

hor...@archildrens.org

archildrens.org

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Re: [Histonet] egg albumin

[Rene J Buesa via Histonet]

Do you have a bottle of "Eggnog"? Egg-albumin will have similar shelf life if 
refrigerated.René 

On Wednesday, April 12, 2017 7:40 AM, Nancy Schmitt via Histonet 
 wrote:
 

 Good Morning-

Thoughts on shelf life of egg albumin?

Thank you

Nancy Schmitt MLT, HT(ASCP)


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Re: [Histonet] Survey!!!!!!

[Rene J Buesa via Histonet]

Staining is a more complex step than coverslipping. Also a coverslip on a 
section has more "latitude" and can, if necessary, be removed and coverslip in 
a different way. A stained section, on the other hand, is almost impossible to 
"correct". Between an autostainer and a coverslipper, I would choose an 
autostainer.René 

On Saturday, April 1, 2017 7:29 PM, Patsy Ruegg via Histonet 
 wrote:
 

 I have seen so many more problems with auto coverslippers, they break down, 
the cover does not hold up over time for archiving, etc., that I would would 
probably chose an autostainer.



Patsy Ruegg, HT(ASCP)QIHC
Ruegg IHC Consulting
40864 E Arkansas Ave
Bennett, CO 80102
H 303-644-4538
C 720-281-5406
prueg...@hotmail.com




From: Ifeoluwa Ajayi 
Sent: Saturday, April 1, 2017 6:11 AM
To: Patti Nelson - PNP Lab Consultant
Cc: Histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] Survey!!

Personally I will prefer auto coverslipper, because cover slipping is
cumbersome and takes time.

Ajayi Ifeoluwa, BMLS, AMLSCN, MSc,
UCH Ibadan, Nigeria.

On Mar 31, 2017 8:00 PM, "Patti Nelson - PNP Lab Consultant via Histonet" <
histonet@lists.utsouthwestern.edu> wrote:

>
> Hi Everyone,
>
> I just wanted to get everyone's opinion. If you had to chose between
> buying a Auto Side Stainer or Auto Slide Cover Slipper, which one would you
> chose? Lets say your volume was around 60 to 80 blocks a day and you worked
> for a GI Lab. Everyone's input would be greatly appreciated.
>
>
>
> Sincerely,
>
> PATTI NELSON  H.T.(ASCP)
> PNP LABORATORY CONSULTANTS
> SUPERVISOR DGC/ZADEH LABS
> PO BOX 412
> CABAZON, CA. 92230
> 909-841-9761
> nelsonr...@verizon.net
> CONFIDENTIALITY NOTICE:This message and any included attachments are from
> Patti Nelson, PNP Laboratory Consultants and are intended only for the
> addressee. The information contained in this message is confidential and
> may contain privileged, confidential, proprietary and/or exemption from
> disclosure under applicable law.  Unauthorized forwarding, printing,
> copying, distribution, or use of such information is strictly prohibited
> and may be unlawful.  If you are not the addressee, please promptly delete
> this message and notify the sender ofthe delivery error by e-mail or you
> may call  909-841-9761.
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Re: [Histonet] Tissue Fixation

[Rene J Buesa via Histonet]

What you describe as a possible scenario is absolutely possible.If your PT does 
not "want to hear" about it, suggest she gets a "hearing aid" or to study 
something about histotechnology or even better yet, pay attention to what a 
professional on the subject (you) has to say about it. You would never dare to 
question her diagnosis, why would she question yours on this subject?René 

On Friday, March 31, 2017 9:11 AM, T H via Histonet 
 wrote:
 

 Good Morning,


I have a pathologist that is not happy with the fixation on some of our LEEP 
specimens.  She swears its histology doing something to the specimen to cause 
the tissue to look unfixed on only "part" of the LEEP specimens (all the same 
client specimens).  She claims we must be diluting our formalin to cause this 
issue or "something".  We mentioned maybe it was on the clients end not placing 
them in 10% formalin right away, she wouldn't hear of it.


Let me give you some back ground on how our process works.  Our clients send us 
all our specimens to us via Overnight FedEx or UPS in 10% formalin they will 
then they sit in 10% formalin in-house until the processors are started around 
3pm and sits an additional 5 hours in 10% formalin on the processor before the 
processor actually starts.  That being said the fixation process has had a 
pretty good start before we ever even touch it.


My question is, and I thought I had read this in the past is, when a specimen 
is left out prior to fixation and lying on a absorbent surface such as a paper 
towel, won't the area of the tissue touching the absorbent surface begin the 
disintegration processes faster in that exact area then the rest of the 
specimen?  Or if you have any other suggestion on what might be happening to 
only "certain" specimens would be great as well.


Thanks for your help!


Tim

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Re: [Histonet] solvent recyclers

[Rene J Buesa via Histonet]

B/R recyclersRené 

On Thursday, March 23, 2017 11:15 AM, Lauren Sweeney via Histonet 
 wrote:
 

 Hi everyone,

We are looking into getting a new solvent recycler to recycle our xylene and 
alcohol, any recommendations?

Thanks!



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Re: [Histonet] low signal for long post fixation

[Rene J Buesa via Histonet]

Perhaps your only solution is to increase HIARRené 

On Thursday, March 23, 2017 9:39 AM, Mariela Chertoff via Histonet 
 wrote:
 

 Dear all,

I have mouse brain tissue perfused with PFA 4% and post fixed in formol 10%
for periods between 2 month and 1 year. I cut them on vibratome (30um) and
I made inmunohistochemistry and beta galactosidase to check senescence.
My problem is that I have less signal on positive controls if postfixation
on formol was longer.

There is a way to solve this problem?

Thanks in advance for your reply

Mariela Chertoff, PhD
Laboratorio de Neuroepigenetica - QB75
 Departamento de Química Biológica
Facultad de Ciencias Exactas y Naturales - UBA
Ciudad Universitaria Pabellón II Piso 4
Ciudad Autónoma de Buenos Aires
C1428EGA - Argentina
Tel: 54 11 4576-3300/09 - Int. 221
email:marielachert...@gmail.com
marielachert...@qb.fcen.uba.ar

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Re: [Histonet] IHC billing on archived case

[Rene J Buesa via Histonet]

You should treat it as a new request.René 

On Wednesday, March 22, 2017 11:25 AM, "Haines, Beth via Histonet" 
 wrote:
 

 Hello all,
After some discussion on IHC billing, I have been asked to verify accepted 
billing practice for the following situation:
An IHC request for 2 single antibody stains has been made by outpatient 
oncology practice on a case (same block) that was completed nearly a year ago. 
6 IHCs were done initially (billed as 88342 & 88341 x 5). How should the new 
request be billed? Should a new visit/encounter be created and 88342 & 88341 be 
billed for this encounter?
Thanks in advance for the guidance.


Beth Haines BS, HTL (ASCP)
bhai...@caperegional.com
Histology Supervisor
Cape Regional Medical Center

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Re: [Histonet] Embedding station

[Rene J Buesa via Histonet]

"Open" your options and try Sakura.René 

On Tuesday, March 21, 2017 3:54 PM, "Flynn, Evelyn via Histonet" 
 wrote:
 

 Hello all,


  Our laboratory is purchasing a new paraffin embedding station.  We are 
considering a Leica Arcadia or a Thermo

HistoStar model.


  Has anyone had good or bad experiences with either of these?


Thanks for your input,

Evelyn Flynn


Lead Research Technologist

Boston Children's Hospital
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Re: [Histonet] need help staining 120um human whole brain sections!

[Rene J Buesa via Histonet]

You have a special project → special tasks so your approach has to be equally 
special.Large brain sections are usually stained while floating but for IH with 
different and successive steps requiring very expensive reagents, floating 
sections is not well suited.You should affix the sections to large slides, and 
I imagine will not be brain whole sections but limited to some areas.In that 
case for IHC you can stain several sections in a humid chamber, manually. I do 
not imagine an automatic system for this task.It will be a costly and slow 
process indeed.René 

On Sunday, March 19, 2017 8:27 PM, Maria Mejia via Histonet 
 wrote:
 

 
Or lab is currently processing a human whole brain.  In about a month or two, 
the whole brain, which will be encased 
in celloidin & serial sections will be cut at 120um each.  Now, we’ve bought an 
old Tetrander cast iron microtome. If 
you haven’t seen one of these microtomes, I can tell you it’s BIG!

Now, we’ll have to stain quite a number of these sections for IHC.  In fact too 
many to handle manually. If possible, need
to find a way to at least stain the majority of these sections in a 
semi-automatic system e.g. washes, quenching & blocking. 

Does any one think it’s possible to convert a LABGO processor (made in India & 
can hold 2 liter glass beakers) or a 
Sakura-Tissue-Tek 4640 (also holds glass beakers - both are old style circular 
processors using glass beakers for staining 
these sections? Does anyone have an alternative system? I could sure use some 
input or ideas anyone welcome
and most appreciated.

Maria Mejia
Lead Histologist
UCSF
Mission Bay
San Francisco, CA




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Re: [Histonet] IHC on alcoholic fixed cytology smears

[Rene J Buesa via Histonet]

M best advise is contacting Dako and obtain from them the IHC manual, which 
covers essentially everything you need to know on this subject.René 

On Saturday, March 4, 2017 2:51 PM, Lynette Pavelich via Histonet 
 wrote:
 

 Hello,
I will soon be starting the process of validating many cytoplasmic,
membranous, and nuclear antibodies on alcoholic fixed cytology smears. They
will be performed using the Bond III.

In doing research on the subject, I am finding many different variables,
and it is starting to be confusing. Is anyone willing to share their
processes, or can suggest good reading material for IHC with alcohol fixed
cytology smears? Hints, tips, tried and true methods would be greatly
appreciated.

Thank you,
Lynette Pavelich, HT(ASCP)
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Re: [Histonet] Fat or Breast & Lipoma Processing Schedule Sought

[Rene J Buesa via Histonet]

You can find all my xylene-free processing schedules at the HistoNet archives. 
They use 2-propanol → mineral oil → paraffin.René  

On Saturday, February 11, 2017 9:44 AM, ian bernard via Histonet 
 wrote:
 

 We use Safe Clear 11 (a Xylene sub) as the clearing agent on our routine
processing schedule; with no intent to resort back to Xylene.  

 

Since we are taking in an influx of lipoma and breast cases, we are seeking
a separate tissue processing schedule.  Thus looking for an overnight
schedule template that we can validate.

 

Are there any histonetters with an overnight schedule that they are willing
to share as our template to begin validation.  Also, any reference source
you can point too along these lines will be greatly appreciated.

 

V/r

IB

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Re: [Histonet] hand staining immunos

[Rene J Buesa via Histonet]

You do not have to "babysit" the procedure for it has well defined/timed steps. 
You just need a timer and check the slides when required. As to procedural "dos 
& donts" try to get a copy of the DAKO IHC manual.René 

On Wednesday, February 8, 2017 7:34 PM, Jennifer via Histonet 
 wrote:
 

 Hello,

 

My dermatopathologist wants me to start hand staining immunos  for MITF. I
would really like some feedback about everyone's experiences/thoughts with
manually hand staining immunos and especially with MITF. Pros and cons? I am
concerned about the amount of time this will take for a tech to babysit the
stain and also about the consistency of the quality in the end result.  Your
opinions are much appreciated.

 

Thank you!

 

Jennifer

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Re: [Histonet] Releasing of Patient Tissue

[Rene J Buesa via Histonet]

Never mind what other people do, just ask your legal department what to do 
because this may involve legal consequences.René  

On Wednesday, January 18, 2017 10:47 AM, Vanessa Keeton via Histonet 
 wrote:
 

 Good Morning All!

I was wondering what everyone's policies were on releasing of patient
tissue other than stones, placentas, and hardware.  Normally we only
release stones, placentas, and hardware but have been receiving request for
other types of tissues lately and was just curious as to what the norm
seems to be pertaining to this issue and if you do release the tissue, how
do you limit patient exposure to blood, body fluids and chemicals?

Thanks!!!

Vanessa Keeton
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Re: [Histonet] Slide and Block Storage

[Rene J Buesa via Histonet]

 We used to keep our blocks on-site during 9 years. During January of each year 
we disposed off 1 year worth of blocks (the oldest) and only kept those deemed 
by the pathologists as interesting for our residents' training 
program.Regarding slides, in 2001 we had 56 years of slides on-site and it was 
decided to send the majority out to a "specialized" facility, and it was a 
complete nightmare. Many slides broke and the final arrangement was incorrect 
becoming extraordinarily difficult finding a case.This situation was most 
likely caused by a poor selection of the storing company. I would never repeat 
this decision but, on the other hand, you could ask: what is the point in 
storing slides for so many years?My suggestion to you is to ask your legal 
department what could be the expectation regarding slides storage, i.e. how 
many years the lab could be liable if a legal suit is levied against the 
hospital? This is a difficult question to answer.Other approach could be to 
follow CAP requirements regarding storage time for blocks and slides. In 
Florida we are required to store blocks for 9 years, and that is what we did.My 
recommendation to you is: keep blocks and slides for the maximum time required 
by your licensing agency, and store them within your actual reach do NOT send 
them out to store outside. Discard the rest keeping only those that represent a 
teaching interest.René   

On Thursday, January 12, 2017 10:59 AM, Amy Self via Histonet 
 wrote:
 

 Happy Thursday connected to Friday,

Have a few questions about slide and block storage...


Are you blocks and slides stored on-site or off-site at a record control type 
facility?

If your blocks and slides are stored off-site how do you get the material that 
you need at any given point of the day?



Thanks in advance for your help,
Amy Self
Histology Lab Senior Tech
Lab
Tidelands Georgetown Memorial Hospital
606 Black River Road
Georgetown, SC 29440
843-520-8711
as...@tidelandshealth.org

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Re: [Histonet] Staining issues

[Rene J Buesa via Histonet]

Improper staining at the center and falling sections are typical consequences 
of poor fixation/infiltration.If you have changed nothing proceduraly, what 
about somebody "new" grossing and preparing thicker tissue slices?René 

On Wednesday, December 28, 2016 8:34 AM, Charles Riley via Histonet 
 wrote:
 

 Our pathologists are complaining that the center of tissues are not
staining properly in both our H's and IHC's. Can anyone provide some
thoughts as to where this problem could be occurring?


As a separate situation we are experiencing the tissues folding or falling
off during the staining process. Both this issue and the staining issue are
recent occurrences and we have not changed our process in any way from
previous years.


Does anyone think these problems are related? Why or why not?  I have
racked my brain with all the trouble shooting techniques I can think of so
any help will greatly be appreciated

-- 

Charles Riley HT(ASCP)CM

Histopathology Coordinator/ Mohs
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Re: [Histonet] Storage temps

[Rene J Buesa via Histonet]

I do not know of anything published other than CAP "requirements" 
(unsubstantiated)René 

On Wednesday, November 23, 2016 9:50 AM, "Richardson, Pam K via Histonet" 
 wrote:
 

 Does anyone know if there is a published acceptable range for storage of 
tissue and paraffin blocks?

Cordially,

Pam ~
+++
Pam Richardson
Clinical Manager
Gundersen Health System Laboratory Services
Email: pkric...@gundersenhealth.org
Phone: 608 775-4133
Fax: 608 775-6136
Interdepartmental Mail Stop: H04-007
E-visit us at: http://www.gundersenhealth.org

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Re: [Histonet] Weigert's hematoxylin for IHC

[Rene J Buesa via Histonet]

Once you finish the IHC procedure, the DAB reaction is very stable and you can 
use Weigert's or any other iron hematoxylin.René 

On Tuesday, November 22, 2016 5:29 PM, Esther C Peters via Histonet 
 wrote:
 

 Could someone advise me on whether Weigert's iron hematoxylin can be used as 
the counterstain for nuclei in IHC? We need to avoid using Harris's hematoxylin 
because that will stain mucocytes in the coral tissues we are working with. If 
it can be used, would that be after all antibody steps have been done? Thank 
you!


Esther Peters


Esther C. Peters, Ph.D.
Term Associate Professor
Environmental Science & Policy
George Mason University
4400 University Drive, MS 5F2
Fairfax, VA 22030-
Office: David King Hall, Room 3050
Phone: 703-993-3462
Fax: 703-993-1066
e-mail: epete...@gmu.edu
http://esp.gmu.edu

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Re: [Histonet] Maximow Method

[Rene J Buesa via Histonet]

Tyrone:A.A.Maximow's Azur-eosin, etc staining produces wonderful results but 
this, and many very old procedures, essentially rest on the use of mercury 
salts which produce special chemical compounds with tissue components.Any, and 
I mean any, deviation from the original procedure will not produce the same 
results as those expected from the original recipes.You can try and you will 
get a staining but without the crisp and delightful colorations obtained with 
the original recipe containing mercury. The same goes for the Harris 
hematroxylin that now is manufactured and sold without mercury oxide and is 
still  named Harris, when really it is not. The results are similar, but not 
exactly as those obtained with the original recipe.So, in my humble opinion, 
you can substitute whatever you want from an original recipe, but please do not 
be surprised if the results do not "live" to your expectations or the original 
description.René  

On Monday, November 21, 2016 2:40 PM, Tyrone Genade via Histonet 
 wrote:
 

 Hello,

I was reading a book from the 60s on the anatomy of aging in man and
animals and the author mentioned using a hematoxylin-eosin Y-Azure-II stain
to show the lymphocytes. Some searching came up with the Maximow Method.
The online protocol I found (for bone marrow):
https://emsdiasum.com/microscopy/technical/datasheet/26252.aspx mentions
the use of Zenker’s or Formalin.

In another old book, Putt's Manual of Histochemical Staining Methods, the
authors says 10% fomalin, Helly's or Zenker's fluid for fixation. I am
definitely not going to start using Zenker's (I get enough grief from the
H officer about picric acid). I normally use Davidson's fixative to fix
and decalsify my fish (formaldehyde, acetic acid + ethanol). Anyone know if
this staining method is compatible with Davidson's fixative?

Would this eosin-Y solution be suitable:
http://www.sigmaaldrich.com/catalog/product/sigma/ht110316?lang=en®ion=US
 for preparing the Eosin-Y Azure-II working solution?

Thanks
-- 
Tyrone Genade
Orange City, Iowa
tel: (+1) 712 230 4101
http://tgenade.freeshell.org

Romans 6:23: The gift of God is eternal life through Christ Jesus our Lord.
To find out how to receive this FREE gift visit http://www.alpha.org.
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Re: [Histonet] Removing Tissue From Tape

[Rene J Buesa via Histonet]

I think your best option is to manually re-coverslip to the slide.René 

On Sunday, October 16, 2016 10:00 AM, Pamela Marcum via Histonet 
 wrote:
 

 Good Morning, 
  
Although the laboratory stopped using tape years ago we are still facing issues 
with "OLD" tape slide being requested for review or moving them to storage only 
to discover the tape has come off and the tissue is stuck to it.  I hesitate to 
just re-coverslip it to the slide by hand.  Can anyone give some ideas of how 
to best preserve these slides that were once taped? 
  
Thank You, 
Pam Marcum 
  
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[Histonet] Fw: Dr. Matsionis

[Rene J Buesa via Histonet]

Hi colleagues:I have just received the sad news that the prestigious Russian 
histopathologist Prof. Alexander Matsionis passed away (see included 
message).Although his name is almost unknown in our field he always was 
extremely enthusiast about new histopathology procedures and helped introducing 
isopropanol tissue processing in scores of Russian labs.My thoughts to his 
family, colleagues and friends.René
 

 On Friday, October 14, 2016 1:58 PM, Maxim Peshkov  
wrote:
 

 Dear Rene!An 12 October was died our great friend, pathologist, doctor 
Alexander Matsionis Academician of RANS (Russian Academy of Natural Sciences), 
MD, PhD, proffesor and teacher. He fought with acute monocityc leikemia. He was 
only 59 years old. He will stay in our memory as a man who made a great 
contribution to the Russian and pathological anatomy as a science (haematology, 
urology, breast pathology, soft tissue and endocrine pathology and many others) 
and as a profession. He organized Rostov pathological-anatomical bureau. He 
also united almost all the pathologists of southern Russia's regions and gave 
way in the life of so many young pathologists, who have already proved 
themselves excellent specialists. He helped to development of our processing 
method with isopropanol and mineral oil. He was a great friend and cheerful, 
always helped the needy both as pathologists, as patients also.He always 
appreciated your help and support.
In our memory and hearts he will stay forever.
Maxim.

   
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Re: [Histonet] new lab

[Rene J Buesa via Histonet]

If you already saw the drawings for the designated area, is it not too late now 
to make changes?If you think you can give input, my only suggestion is that you 
set your working areas in a way that they follow the workflow and ideally 
should be close to the surgery suits.René 

On Thursday, October 13, 2016 12:47 PM, "Blazek, Linda via Histonet" 
 wrote:
 

 Hi all,

It's finally official.  We will be building a new facility with ground breaking 
in the spring!  I have seen the drawings for the designated area in the new 
building for the histology lab.  Now I could use any suggestions that you all 
may have in designing a new lab.

Thanks in advance!
Linda

Linda Blazek HT (ASCP)
Pathology Lab Manager
GI Pathology of Dayton
Digestive Specialists, Inc
Phone: (937) 396-2623
Email: lbla...@digestivespecialists.com


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Re: [Histonet] Buffered formalin substitution

[Rene J Buesa via Histonet]

Julio:Unfortunately NBF is the OVERALL best fixative there is. ANY substitute 
will be good for some things and not that good for others. Under those 
circumstances what to do? Simply use LESS amounts of formalin, do it safely 
keeping to a minimum its exposure.Under separate cover I am sending you 2 
articles I published on the subject.René 

On Friday, September 30, 2016 8:04 AM, Julio Benavides via Histonet 
 wrote:
 

 Hi there,

The health officer of our institute has risen (again!) the issue of  
substituting buffered formalin for some other less hazardous fixative .

I would like to know you opinion, and experience, in such matter. Is  
it even possible? Have you successfully done it? In that case, which  
samples you normally handle? We are a research institute, doing  
several ruminant PMs a week, so big chunks of tissue to fix. Then, we  
normally do IHCs with a variety of antibodies, depending on the  
project.  My worries are that, in case buffered formalin could be  
substituted, you can never be sure about IHC and maybe antibodies that  
were working before stop doing it.

Very many thanks for your thoughts and time in this issue. Greatly  
appreciated!

Cheers

Julio



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Re: [Histonet] Egg Whites and IHC Staining

[Rene J Buesa via Histonet]

Egg white (in Mueller's albumin) will always produce a "shadowy" staining with 
IH procedures y a dark background with IHC procedures.I suggest you use pork 
gelatin dissolves in the water of the water bath, use (+) charged slides and 
increase the drying time in the oven.Also sectioning those large pieces as thin 
as possible will help.René 

On Monday, September 12, 2016 10:25 AM, Jessica Riggleman via Histonet 
 wrote:
 

 Hello,

We are cutting large Rabbit PLF formalin fixed paraffin embedded sections.

We have had issues staining H and Goldner’s Trichrome. Egg whites is working 
as a great slide adhesive for these two, but not so much for IHC Staining (RAM 
11 specifically).

Any ideas/suggestions on how to get these sections to adhere? We have tried 
many methods, such as positively charged slides, chrom-alum gel, and so on.

Thank you for all input,
Jessica


_

Jessica Riggleman | Research Associate

Globus Medical, Inc.
Valley Forge Business Center
2560 General Armistead Avenue | Audubon, PA 19403
Ph: (610) 930-1800 ext. 2583 | Fax:

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Re: [Histonet] Modified Movat's Pentachrome stain

[Rene J Buesa via Histonet]

Do you have any contacts at any old histology lab, i.e., one that has been in 
operation for more than 50 years? You may find there iodine crystals and ask 
for a few grams. I used to have a 500 g bottle at my lab (which began in 
1947).René 

On Thursday, September 1, 2016 10:39 AM, Angela Lamberth 
<alambe...@lji.org> wrote:
 

 I would prefer to not substitute anything yet the procedure calls for a 
minimum of 2 grams of iodine which I simply cannot buy in the United States 
thanks to new DEA regulations.
On Thu, Sep 1, 2016 at 7:21 AM, Rene J Buesa <rjbu...@yahoo.com> wrote:

Once you start substituting things in an original recipe, the outcome cannot be 
expected to be what the original recipe was supposed to deliver.Iodine crystals 
cannot be substituted by Lugol because, besides the iodine also contains its 
salts. and alcohol. They are two completely different things. This is the same 
as using Harris hematoxylin without mercury, you can keep calling it Harris, 
but it no longer will stain the same. Finally I do not see any advantages of 
Movat's pentachrome over a good Masson. Just my opinion but I think you are 
embarking in a complex process with little chance of finishing with the 
expected results.René 

On Wednesday, August 31, 2016 9:53 PM, Angela Lamberth via Histonet 
<mailto:histonet@lists.utsouthwestern.edu> wrote:
 

 Hi Histonetters!

I’m gearing up to perform a pentachrome stain. I will be making this in
house and not using a kit. Through searching histonet, I’ve found a
protocol used by the Children’s Hospital of Philadelphia Pathology Core.
http://pathcore.research.chop.edu/docs/MovatPentachromeStain.pdf


Two things:  Iodine crystals are out of the question. Can I replace that
with Lugol’s iodine solution or should I just substitute Weigert’s for the
hematoxylin in this protocol?


Also, saffron vs tartrazine vs Orange G stain.  There is a real difference
in price. Does anybody have a personal preference in terms of quality or
aesthetics? Is paying the extra money for saffron really worth it?


Thanks!

Angela



-- 
Angela Lamberth
Histology Technician II
Histology Core Lab
La Jolla Institute for Allergy & Immunology
9420 Athena Circle
La Jolla, CA 92037
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-- 
Angela LamberthHistology Technician IIHistology Core LabLa Jolla Institute for 
Allergy & Immunology9420 Athena CircleLa Jolla, CA 92037

   
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Re: [Histonet] Modified Movat's Pentachrome stain

[Rene J Buesa via Histonet]

Once you start substituting things in an original recipe, the outcome cannot be 
expected to be what the original recipe was supposed to deliver.Iodine crystals 
cannot be substituted by Lugol because, besides the iodine also contains its 
salts. and alcohol. They are two completely different things. This is the same 
as using Harris hematoxylin without mercury, you can keep calling it Harris, 
but it no longer will stain the same. Finally I do not see any advantages of 
Movat's pentachrome over a good Masson. Just my opinion but I think you are 
embarking in a complex process with little chance of finishing with the 
expected results.René 

On Wednesday, August 31, 2016 9:53 PM, Angela Lamberth via Histonet 
 wrote:
 

 Hi Histonetters!

I’m gearing up to perform a pentachrome stain. I will be making this in
house and not using a kit. Through searching histonet, I’ve found a
protocol used by the Children’s Hospital of Philadelphia Pathology Core.
http://pathcore.research.chop.edu/docs/MovatPentachromeStain.pdf


Two things:  Iodine crystals are out of the question. Can I replace that
with Lugol’s iodine solution or should I just substitute Weigert’s for the
hematoxylin in this protocol?


Also, saffron vs tartrazine vs Orange G stain.  There is a real difference
in price. Does anybody have a personal preference in terms of quality or
aesthetics? Is paying the extra money for saffron really worth it?


Thanks!

Angela



-- 
Angela Lamberth
Histology Technician II
Histology Core Lab
La Jolla Institute for Allergy & Immunology
9420 Athena Circle
La Jolla, CA 92037
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Re: [Histonet] I guess I've found it at last

[Rene J Buesa via Histonet]

Yes, I received it.Most probably it is a disguised "junk/spam" 
advertisement.Just in case do NOT open it.René 

On Wednesday, August 24, 2016 10:58 AM, "Macke, Gail via Histonet" 
 wrote:
 

 Histonet,
Received this today.
What is this?
Can you look into this?
Has anyone else received this?
Took off “nowakc” from email address and Marcie DuVernay.


Gail Macke,HTL
On Aug 23, 2016, at 7:53 PM, nowakc via Histonet 
> 
wrote:

Hey friend,

As you know I've  been looking for some  suff for a long time, and I think I've 
 found it at  last, just take  a look 


Faithfully, nowakc


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Re: [Histonet] Quantifiable evaluation criteria

[Rene J Buesa via Histonet]

1- Make a list of ALL the tasks you delegate on this "Lead Histo"2- Quantify 
each tasks, i.t. give a "numeric weight" to each  of a maximum 100 points.3- 
Keep track of how the "Lead Histo" performs in each and DISCUSS your evaluation 
with the "Histo Lead" quarterly. This will allow the "Lead Histo" know how you 
appreciate his work and which tasks need improvement.4- Come the annual 
evaluation just add-up all the points against a maximum allowable of 100% and 
you will get an unbiased/discussed evaluation of your "Lead Histo".That is how 
I used to do it.René 

On Wednesday, August 24, 2016 10:36 AM, Steven Crochiere via Histonet 
 wrote:
 

 How would one develop an evaluation standard for a "Lead Histo" that is more
quantifiable rather than subjective criteria? I cannot see it as far as
"leadership" skills.

Any suggestions?

 

Thanx,

 

steve

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Re: [Histonet] Query on authorship

[Rene J Buesa via Histonet]

Jorge:The first thing is to be absolutely sure the data is worth publishing and 
that the results have scientific relevance.If this is the case and both you and 
the other contributor agree I think the data should be published.In no way you 
should eliminate the data obtained by the other individual you have been unable 
to locate because that will weaken the results. If what the two students did 
was obtaining data it is fair to add the name of the one accepting to be 
considered as co-author, but ONLY if s/he takes part of data processing and the 
conclusions.As to the other student you have been unable to locate I do NOT 
think you should make him/her as co-author because just obtaining raw data does 
not amount to that, nor it will be fair to the one you will include as 
co-author if co-authorship includes processing/analyzing the data.The fairest 
thing to do is to mention the other student in the acknowledgements very 
clearly pointing out his/her actual participation.This is how I would solve 
this issue. On the other hand I have never consider as "co-author" of any of my 
papers any person whose only task was gathering information or following a 
field protocol to obtain data or make experiments.René 

On Friday, August 19, 2016 3:50 PM, Jorge A. Santiago-Blay via Histonet 
 wrote:
 

 Query on authorship

Dear Colleagues:

I am writing a small paper resulting from research done with two
undergraduates many years ago (and, later on, involving several other
colleagues using cutting-edge technology). As the results became obvious,
both of the students agreed (orally, in person) with me that we should get
the research published. As far as I remember, there was no email or letter
documenting that and, there was no manuscript, only the data and the
methods we were using.

The problem: I have located one of the former students (now a researcher at
a major research institution), who is excited about getting the research
published, but not the second student.

Question: How to handle the contribution (including authorship) of the
other person? Here are some options I see.

a. *Omit the name of the person that has not been located* and indicate
that another person was involved in the data collection but we were hot
able to locate him/her to get his/her approval to use his/her name as an
author.  Under these circumstances, would it be OK to name the person in
the Acknowledgments? Lately, I am asking permission to do that because
sometimes some people prefer to remain anonymous.

b. *Include the name of the person I cannot locate as an author*, an act of
fairness and good faith on my part. If the person does not like the idea
(and the paper is published) retract the name of the person in an erratum,
later on, and assume responsibility for my error. A kind colleague did that
to me once and, subsequently, it has resulted a long standing collaboration
(and co-authorship in many papers, with my knowledge) :)

c. *Nor use the data garnered by the person I cannot locate*. Although I am
pretty sure I am authorized by the institution to use the data, as a
general personal; preference, I like to ask permission.

If you have something constructive to comment, kindly direct your comments
to me, blayjo...@gmail.com ,

Apologies for potential cross-posting.

Sincerely,

Jorge

Jorge A. Santiago-Blay, PhD
blaypublishers.com

1. Positive experiences for authors of papers published in *LEB*
http://blaypublishers.com/testimonials/

2. Free examples of papers published in *LEB*:
http://blaypublishers.com/category/previous-issues/.

3. *Guidelines for Authors* and page charges of *LEB*:
http://blaypublishers.com/archives/ *.*

4. Want to subscribe to *LEB*? http://blaypublishers.com/subscriptions/

http://blayjorge.wordpress.com/

http://paleobiology.si.edu/staff/individuals/santiagoblay.cfm
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Re: [Histonet] oil red in WAT frozen samples

[Rene J Buesa via Histonet]

Apply gently heated water on the sections in a way that the gelatin is washed 
out.René 

On Tuesday, August 16, 2016 4:48 AM, Monica Aguilera via Histonet 
 wrote:
 

 Dears,

I was wondering if some of you might have experience in the following:

We have had a lot of problems cutting WAT tissue either directly embedded
in OCT or pre-fixed with PFA or formalin (either cold or and them embedded
with OCT.
We solved this problem following Ryan Berry et al, 2014 paper protocol with
gelatin.
We get really nice cuts with that, but when performing the staining with
oil red, we can not remove the gelatin from the cut and the oil red stays
on the top of the tissue cut, it looks like there an oil-aqueous phase the
does not let the oil red enter into the tissue

Any suggest for removing the gelatin and for letting the oil red entering
into the tissue will be more than welcome!

Many thanks and kind regards,

Mònica

-- 
Mònica Aguilera Pujabet, DVM, PhD
Histopathology Facility
Institute for Research in Biomedicine - IRB Barcelona
Baldiri Reixac, 10
E-08028 Barcelona - Spain

Tel:  +34 934033776 <%2B34%20934020546>
monica.aguil...@irbbarcelona.org 
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Re: [Histonet] Antibodies storage tips

[Rene J Buesa via Histonet]

I used to have several compartmented plastic alphabetized boxes with enough 
empty spaces to accommodate "new arrivals". If the spaces were used-up I just 
added a new box. Since they occupied several shelves, each shelf had the 
lettering identifying the boxes in each one.René 

On Tuesday, August 16, 2016 6:29 AM, Julio Benavides via Histonet 
 wrote:
 

 Hi all,

In our lab we carry out quite a number of IHCs for research purposes. 
The number of mAbs and pAb stored in the frezeer is getting to the point 
that, when you need to do and IHC, you actually lose more time diving 
through the frezeer draws for the right vial than doing the IHC! is not 
a question than we are running our of freezer room, it is more to the 
point of having a proper and standarized way to keep the antibodies and 
than produce a proper registry of what we have and where to find it.

I was wondering, in an attempt to avoid reinventing the wheel, if you 
could be as kind as to share your tips for storing and organizing the 
antibodies (aliquots, stock solutions, intermediate dilutions) so 
chaos does not take over the lab!

As always, thank you very much for your time and advice!

Cheers

Julio


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Re: [Histonet] Placing formalin on specimens

[Rene J Buesa via Histonet]

Usually you do not pour 10%NBF onto a specimen; you place the specimen onto a 
container/vial with the 10%NBF.René 

On Tuesday, August 16, 2016 9:29 AM, Mike Pence via Histonet 
 wrote:
 

 I know this might sound a bit crazy, but does anyone have a written policy and 
procedure for dispensing 10% formalin onto a surgical specimen? If you do, 
would you be willing to share?

Michael S. Pence | Supervisor of Laboratory Services
Great River Health Systems
1221 S. Gear Ave. | West Burlington, IA 52655
Office 319-768-4546 | Main 319-768-4525 | Fax 319-768-4557
mpe...@grhs.net | 
www.greatrivermedical.org.
www.Facebook.com/GreatRiverHealthSystems
 | www.Twitter/GreatRiverMed


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Re: [Histonet] specimen storage cabinets

[Rene J Buesa via Histonet]

Get regular metal cabinets used to store garage items.They are sold at any 
general store (such as HomeDepot or Walmart).René 

On Thursday, August 11, 2016 12:05 PM, Atoska Gentry via Histonet 
 wrote:
 

 Hello, I work for a research facility and we currently have an archive of 
several formalin fixed samples stored in 500ml glass specimen jars; one of our 
PI's recently asked me to conduct a search for storage cabinets specifically 
designed to accommodate these samples. Is anyone aware of the availability of  
such cabinets? The only thing my search has turned up thus far are the 
flammable metal cabinets. But due to limited storage space we have to stack 
these jars and the generic flammable cabinets are not stacked glass specimen 
jar suitable. All ideas & input are welcome. Thank you kindly, Atoska
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Re: [Histonet] On the hunt for new microtomes!

[Rene J Buesa via Histonet]

As far as I know Leica Microsystems has bought/absorbed along the years many 
optical/instruments makers, such as Baush, American Optical, Cambridge 
Instruments, Wild-Heerburg (Switzerland, although this one was a "reverse" 
acquisition), Australian "Bond" manufacturers, NOVOCASTRA laboratories BUT I 
have never heard that it has bought Sakura instruments. It would be nice if 
somebody has reliable information about this alleged acquisition.René  

On Monday, August 1, 2016 4:02 PM, Rene J Buesa via Histonet 
<histonet@lists.utsouthwestern.edu> wrote:
 

 I don't know now, but some years ago Thermo instruments were less that 
reliable. Try Leica or even better Sakura.René 

    On Monday, August 1, 2016 3:20 PM, Mary Faith Encarnacion via Histonet 
<histonet@lists.utsouthwestern.edu> wrote:
 

 Hey HistoNet,

Thanks to everyone who helped me out by providing their opinions on
embedding centers.  This time, I need everyone's thoughts on microtomes.
My lab is debating between the Thermo/Microm HM355S and the Leica RM2255.
Your thoughts and advice are very much appreciated!  If there are any more
I should try, let me know!

Thanks again,
Mary Faith
Histology Supervisor
VA Palo Alto Health Care System
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Re: [Histonet] On the hunt for new microtomes!

[Rene J Buesa via Histonet]

I don't know now, but some years ago Thermo instruments were less that 
reliable. Try Leica or even better Sakura.René 

On Monday, August 1, 2016 3:20 PM, Mary Faith Encarnacion via Histonet 
 wrote:
 

 Hey HistoNet,

Thanks to everyone who helped me out by providing their opinions on
embedding centers.  This time, I need everyone's thoughts on microtomes.
My lab is debating between the Thermo/Microm HM355S and the Leica RM2255.
Your thoughts and advice are very much appreciated!  If there are any more
I should try, let me know!

Thanks again,
Mary Faith
Histology Supervisor
VA Palo Alto Health Care System
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Re: [Histonet] Feedback on Immunohistochemistry stainers

[Rene J Buesa via Histonet]

Request a DAKO demo.René 

On Thursday, July 28, 2016 8:01 PM, Gmail via Histonet 
 wrote:
 

 
Hi all,
We are looking into getting a new staining platform for our IHC lab. I would 
appreciate any feedback from your experience regarding ease of use, how long 
maintenance;daily , monthly takes to perform and through put. Are there any 
issues you ran into ? We run approximately 4-6000 slides/ month and are looking 
into Roche, Leica and Dako.
Thanks in advance


Andrea Beharry
Senior MLT, Immunohistochemistry 
William Osler Health System




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Re: [Histonet] block scrapers

[Rene J Buesa via Histonet]

Use a regular one blade pocket knife (as I used to do).René 

On Thursday, July 21, 2016 2:49 PM, Lauren Sweeney via Histonet 
 wrote:
 

 Hi all,

My lab is in need of some tools to scrap the paraffin off the edges of the 
blocks after embedding. Does anyone have any recommendations for me?


Thanks!!
L
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Re: [Histonet] Reticulin Stain

[Rene J Buesa via Histonet]

Gomori's René 

On Thursday, July 21, 2016 12:04 PM, Anita Buchiane via Histonet 
 wrote:
 

 Does anyone out there still do the retic by hand?  If so, can you share which 
procedure you use?  Thanks




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Re: [Histonet] Cleaning Tissue Molds

[Rene J Buesa via Histonet]

Place your molds in a 2% dishwasher soap boiling solution for 5 minutes → was 
in running water for 5 minutes → dry in a convection oven at 60ºC for 10 
minutes and your molds will be ready to use.As a "release" solution use a 
mixture 1:1 of 2-propanol and mineral oil (light weight).René 

On Thursday, July 14, 2016 6:34 AM, "Kennedy, Lisa via Histonet" 
 wrote:
 

 Dear Fellow Histo Techs,
What is the BEST practice for cleaning the paraffin block tissue molds?  We do 
not want to use our processor due to its age and wear and tear and frequent 
replacement of cellanoids when we clean them via the processor.
Thanks so much for your help! In advance.
Sincerely,
Lisa Kennedy, HT (ASCP)

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Re: [Histonet] curiosity may kill me

[Rene J Buesa via Histonet]

I always used Auramine at room temperature to identify TB bacilli in tissue 
sections with fluorescence filter, never used heat and the results were as 
expected. Bancroft is the one describing the procedure using heat for 
auramine/rhodamine procedure but auramine alone at room temperature is 
enough.René 

On Tuesday, July 5, 2016 4:24 PM, "Bitting, Angela K. via Histonet" 
 wrote:
 

 I'm curious as to why we heat the Auramine Rhodamine when we are staining 
tissue sections but our Micro lab stains at room temp.  It is because "it's 
always been done that way"?
Thanks group,
Angie






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Re: [Histonet] Certification

[Rene J Buesa via Histonet]

Your tech has an "above deserved" expectation.How would you even consider 
promoting somebody who is not qualified to even be HT certified to Lead 
HT/Coordinator?This is disrespectful for those who reversing that position have 
been unable to achieve it.It speaks volumes about your tech aspirations and 
your managerial skills.René
 

On Friday, July 1, 2016 1:50 PM, Terri Braud via Histonet 
 wrote:
 

 Respectfully, a public forum is no place to discuss  personnel issues on such 
a personal level.  

Terri L. Braud, HT(ASCP)
Anatomic Pathology Supervisor
Laboratory
Holy Redeemer Hospital
1648 Huntingdon Pike
Meadowbrook, PA 19046
ph: 215-938-3689
fax: 215-938-3874

  4. Certification (Adesupo, Adesuyi (Banjo))

--

Message: 4
Date: Fri, 1 Jul 2016 09:45:31 -0500
From: "Adesupo, Adesuyi (Banjo)" 
To: "'histonet@lists.utsouthwestern.edu'"
    
Subject: [Histonet] Certification
Hi,
    I hope you guys are doing great. Please I have a question and would 
appreciate your contributions. I have a tech that could not pass the 
certification test, when the ASCP were still accepting high school diploma for 
HT.
The tech is no longer eligible to take the test again, because he did not have 
the minimum requirement (i.e. Associate Degree) to register for the test. He 
wants me to promote him to Histology Lead Tech/Histology Coordinator.
What do you guys think about this?

Best regards,

Banjo Adesuyi, BMLS, HT (ASCP) HTL, QIHC, QLS
Histology Supervisor
Norman Regional Health System,
Norman, OK 73071.
Tel: 405- 307- 1145
Cell: 405-973-6363

=
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Re: [Histonet] How to get your stains more vibrant when cutting at 2μm?

[Rene J Buesa via Histonet]

Angela:"Pale" results are the trade-off for great quality very thin "2 µm" 
sections but you can always improve intensity somewhat .1- your "regressive" 
stain, if it is "modern Harris" has the inherent problem of lacking mercury 
chloride and it is little you can do about. Perhaps if you use "progressive 
Mayer" you will get better results. You will not have to differentiate (with 
the intrinsic "danger" of leaving the section too pale) and if used fresh 
Mayer's can be a good approach.2- as to the counterstain perhaps you should add 
safranine to the eosin (20% safranine + 80% eosin) and will get a darker red.3- 
try to dehydrate as quickly as possible or even better, wash the sections in 
distilled water and place them in an oven at 60ºC for 10 minutes and coverslip 
as usual. You will eliminate any "color wash" due to the alcohols. If you've 
not enough "trust" on dry/oven dehydration, try with some sections as a test. 
You will like the method.René 

On Tuesday, June 28, 2016 5:53 PM, Angela Lamberth via Histonet 
 wrote:
 

 When I cut at 2μm my H and special stains look pale. How can I get my
stains to pop or am I stuck with pale looking stains when sectioning that
thin?

I run manual specials and a manual regressive H For H I've tried
increasing my time in hematoxylin (beyond the manufacturer recommendation),
diluting my acid alcohol differentiation, and increased time in eosin but
the slides still lack the vibrancy that many of the postdocs desire.

I use Shandon instant hematoxylin and alcoholic eosin by Thermo. Everything
else I prepare in house from scratch. Any recommendations?
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Re: [Histonet] Spill kits

[Rene J Buesa via Histonet]

What you need to do is to communicate to everybody where the kits are, and 
place them where it is more convenient for you. Once everybody knows the 
location, a good sign is always a plus.René 

On Tuesday, June 14, 2016 10:15 AM, Anne Murvosh via Histonet 
 wrote:
 

 Do spill kits need to be out where anyone can see them, or can I put it in a 
cupboard with a sign on the door?  I need to move mine and there's very little 
room.  Thanks Anne
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Re: [Histonet] Over-processing of brain tissue

[Rene J Buesa via Histonet]

It seems to me you are processing too much unless the slices are 3mm thick or 
more.I suggest you to cut the dehydration to 45 minutes (the sequence seems 
OK)Reduce the pure 2-propanol to just 2 changes (30 min is OK)Add 1 change of a 
mixture 1:1 of 2-propanol and xylene + 2 xylene stepsthen to paraffinUse vacuum 
and mixing/agitation in all steps (it will help)If you want really to simplify, 
use 2-propanol in all steps instead of ethanol and eliminate the xylene. You 
can go from pure 2-propanol → 1:1 mixture of 2-propanol with paraffin → 3 
paraffin changes.You will eliminate xylene and obtain very good results.Contact 
me if you need some publications on this procedure.René 

On Tuesday, June 14, 2016 9:53 AM, "Wheelock, Timothy R. via Histonet" 
 wrote:
 

 Good morning everyone:

I seem to be having problems with over-processing of brain tissue.
I use a VIP6 processor.
I start off with 1 hour each of 30%, 50%, 80%, and 95% isopropanol.
Then half-hour each of three changes of absolute isopropanol.
Then half hour each of three changes of xylene.
Then half-hour each of 4 changes of Paraplast.

I use a slow mixing cycle and no vacuum-pressure for all reagents except the 
paraffin.
For the paraffins, I use vacuum pressure, but no mixing cycle.

I rotate the paraffins during each run of tissue.
I replace the dilutions of isopropanol after I have processed 500 blocks.
I rotate the absolute isopropanols and xylenes after 500 blocks as well.

When I embed the brain tissue, it sometimes seems a little stiff, or even a bit 
brittle, to one extent or another.
When I trim the blocks, they seem somewhat dry. Once in a while there is even a 
saw-dust effect.
Before I section the blocks, I have to keep them on ice for at least 3 hours 
before they are moist enough to cut.
Even then, the first case (out of 5 cases) shows chatter in the cerebral cortex.
When I examine the stained sections microscopically, even the best sections 
look a little "rough" or dry.
Taking microscopic images can be difficult at 40x ( or even 20x) because all 
this roughness shows.
The brain tissue looks "granular" rather than smooth, especially with a LHE 
stain.

I have continued to reduce the times to their present values, but it still does 
not seem enough.
I absolutely love the VIP6, but it is a much more powerful machine than the 
Shandon Hypercenter XP that I use to have.
Perhaps I have still not fully compensated for this power.
It has 4 paraffin reservoirs rather than 2 on the old Hypercenter, and the 
reagent reservoirs hold twice as much volume.

Any ideas on how to resolve this problem?
Should I reduce the times further?
Should I alter the use of the mixing and/or vacuum-pressure?

Thanks for any help that you can give me.

Tim


Tim Wheelock
Assistant Director, Neuropathology
Instructor of Neuroanatomy
Tour Coordinator
Harvard Brain Tissue Resource Center
Room 203, Mailman Research Center
McLean Hospital, Belmont, MA 02478
Phone: 617-855-3592
Cell:    857-234-9311
Fax:    617-855-3199



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Re: [Histonet] Powder Picric Acid vs Liquid Picric Acid

[Rene J Buesa via Histonet]

If your 1.3% picric solution is in distilled water, there is not much you can 
do about it.If it is in acetone 38.4 mL contains 0.5 g of picric acid to which 
you can add 361.6 mL of acetone to get your desired concentration.René 

On Monday, June 13, 2016 3:24 PM, Jennifer MacDonald via Histonet 
 wrote:
 

 0.5 grams in 400 mL is 0.125%.  Dilute your 1.3% tenfold and it will be 
close enough.



From:  Amy Johnson via Histonet 
To:    "histonet@lists.utsouthwestern.edu" 

Date:  06/13/2016 10:47 AM
Subject:        [Histonet] Powder Picric Acid vs Liquid Picric Acid



Our current procedure for the Brown and Brenn Gram stain uses 0.5 grams of 
picric acid to 400mls of acetoneis there a way to use a 1.3% 
Picric Acid solution in place of the powdered Picric Acid?

Amylin Johnson, B.S. HTL(ASCP)
Associates in Pathology
Wausau Wi 54401
715-847-2130

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Re: [Histonet] retaining cut slides to keep or not to keep

[Rene J Buesa via Histonet]

It seems you have "baked" and "unbaked" slides.4ºC storage is always more 
expensive and "baked" slides are keep very well at RT.I think your first step 
is to ask around who would like those specific slides.If they will be used in 
the future, "bake" those "unbaked" and store all at RTRené 

On Monday, June 6, 2016 1:45 PM, Rachel M Gonzalez via Histonet 
 wrote:
 

 Hi everyone,

I work in a research lab and have a number of slides that have been cut by
the previous person in this position. Several slides are over one year.  I
would like to know what is the standard practice for keeping cut slides? I
have used them and they stain just fine but the question was brought up in
a lab meeting.

How long can you keep unbaked slides? Room temperature/4C

How long can you keep baked slides? Room temperature/4C

Thanks
Rachel Gonzalez PhD
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Re: [Histonet] Histology tips

[Rene J Buesa via Histonet]

Would you share what you receive for the amusement/benefit of us all?René 

On Tuesday, May 31, 2016 9:15 AM, Charles Riley via Histonet 
 wrote:
 

 I am trying to do a histology tip of the week for my new histo team as a
way to help them learn some new ways to do troubleshooting. I've run out of
ideas at the moment. If anyone has any fun, interesting, or extremely
useful tip for troubleshooting any issues please send them to me. I would
really appreciate it.

I also just want to add in a thank you for all the help I have received
from all our users. Everyone has been so helpful and I have been able to
fix a large amount of errors my predecessors has left piled up for me to
fix.

-- 

Charles Riley HT(ASCP)CM

Histopathology Coordinator/ Mohs
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Re: [Histonet] Pap stain without xylene

[Rene J Buesa via Histonet]

Hi Mike:The steps you desrcibe are wrong.After you finish staining your PAP 
smear, just wash them in the last ethanol → oven dry at 60ºC for 5 minutes or 
as required if the smear is too thick and when completely dried → 
coverslip.This final drying has to take place at temperatures above room temp. 
because you have to be absolutely sure the smear (or stained tissue section)  
is absolutely dried before applying the mounting medium → coverslip.The steps 
you write about 2-propanol and mineral oil are those the tissues have to go 
through before paraffin infiltration. René 

On Tuesday, May 24, 2016 6:36 PM, Mike Toole via Histonet 
 wrote:
 

 René and other Histonetters,


After reading the paper:



Buesa RJ, Peshkov, MV. Histology without xylene. Ann Diagn Pathol. 2009 
Aug;13(4):246-56. Epub 2009 Feb 5.



It appears that xylene in the final clearing steps is replaced with isopropanol 
and mineral oil as follows:

·        5:1        isopropanol to mineral oil            50°C

·        2:1        isopropanol to mineral oil            50°C

·        Undiluted mineral oil                                    50°C

·        Drying oven 5 minutes                                  60°C

·        Coverslip



Do you feel these clearing steps be applied to the pap stain in order to 
eliminate xylene?  If so, can it be done at room temperature?

Thanks,
Mike

Mike Toole, BS, CT(ASCP)CM

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Re: [Histonet] vibrations

[Rene J Buesa via Histonet]

Photomicrography could be affected at high resolution (immersion oil 
objectives) but probably could be eliminated if the microscope table is 
isolated from the floor with some vibration damping device.René 

On Tuesday, May 17, 2016 9:52 AM, Terri Braud via Histonet 
 wrote:
 

 With experience and certification in both, I think that problems from ambient 
vibrations are much less severe in Histology than EM.  There are special 
vibration dampening worktables, often used for EM, that would eliminate the 
problem for cutting stations, or any other sensitive equipment, and still allow 
you to use the space.  I hope this helps.  Terri

Terri L. Braud, HT(ASCP)
Anatomic Pathology Supervisor
Laboratory
Holy Redeemer Hospital
1648 Huntingdon Pike
Meadowbrook, PA 19046
ph: 215-938-3689
fax: 215-938-3874

-Original Message-
From: histonet-requ...@lists.utsouthwestern.edu 
[mailto:histonet-requ...@lists.utsouthwestern.edu] 
Sent: Monday, May 16, 2016 1:00 PM
To: histonet@lists.utsouthwestern.edu
Subject: Histonet Digest, Vol 150, Issue 18

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Contents of Histonet digest..."


Today's Topics:

  1. floor vibration (Nancy Schmitt)
  2. Re: floor vibration (Paula Keene Pierce)


--

Message: 1
Date: Mon, 16 May 2016 15:03:25 +
From: Nancy Schmitt 
To: "'histonet@lists.utsouthwestern.edu'"
    
Subject: [Histonet] floor vibration
Message-ID: 
Content-Type: text/plain; charset="us-ascii"

Happy Monday!
We are moving to a new space and part of our area is above the laundry - there 
is some vibration from there.  Does anyone have any experience with this and 
could you please share how you accommodated this?  Special flooring, pads, etc.
Thank you much!

Nancy Schmitt HT, MLT (ASCP)


Message: 2
Date: Mon, 16 May 2016 15:29:35 + (UTC)
From: Paula Keene Pierce 
To: Nancy Schmitt ,     Histonet
    
Subject: Re: [Histonet] floor vibration
Message-ID:
    <1798931453.3273285.1463412575143.javamail.ya...@mail.yahoo.com>
Content-Type: text/plain; charset=UTF-8

Refuse the space and ask to be moved somewhere else.
Years ago our EM scopes were on the fourth floor and lines in photos taken 
could be seen from the vibration from people simply walking down the halls.
Finally moved the scopes to the basement on a section of concrete cut out 
completely separate from the rest of the building.?Paula Keene Pierce, BS, 
HTL(ASCP)HTPresidentExcalibur Pathology, Inc.5830 N Blue Lake DriveNorman, OK 
73069PH 405-759-3953FAX 405-759-7513www.excaliburpathology.com

      From: Nancy Schmitt via Histonet 
 To: "'histonet@lists.utsouthwestern.edu'" 
 Sent: Monday, May 16, 2016 10:03 AM
 Subject: [Histonet] floor vibration
  
Happy Monday!
We are moving to a new space and part of our area is above the laundry - there 
is some vibration from there.? Does anyone have any experience with this and 
could you please share how you accommodated this?? Special flooring, pads, etc.
Thank you much!

Nancy Schmitt HT, MLT (ASCP)




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Re: [Histonet] Melanin Bleach

[Rene J Buesa via Histonet]

You are right. Bleaching is a "rough" procedure for the "survival" of sections 
and if on top of that you left the section overnight in DiH2O that is a recipe 
for disaster, as the one you experienced. Try to do the whole procedure during 
the same day.Additionally it seems to me that 6h in potassium permanganate is 
too much, you should check the condition of the sections every hour trying to 
minimize KMnO4 action the least time possible.René 

On Tuesday, May 10, 2016 12:10 PM, "Heckford, Karen - SMMC-SF via Histonet" 
 wrote:
 

 Good Morning,
I need some help.  Yesterday I bleached some heavily pigmented tissue.  I have 
to run some IHC's on them.  I bought the bleaching kit from American Master 
Tech.  I had to put the Permanganate for about 6 hours to get the melanin and 
then a couple of minutes in Oxalic Acid.  I had to let them set overnight 
because I could not get another IHC run in that day.  It looks like the tissue 
fell off during decloaking.    I used Apex slides.  I rarely ever have to 
bleach anything here.  So I am not sure if I did this correctly.  I am thinking 
I need to bleach and do the run in the same day and not let them set over night 
in DiH20.

Thanks,


Karen Heckford HT ASCP CE
Lead Histology Technician
St. Mary's Medical Center
450 Stanyan St.
San Francisco, Ca. 94117
415-668-1000 ext. 6167
karen.heckf...@dignityhealth.org
                                                                                
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Re: [Histonet] Cytology/Histology Staining Question

[Rene J Buesa via Histonet]

I would be concerned with potential cross-contamination. In my lab we had 2 
staining instruments, one for cytology and other for histology.René 

On Tuesday, May 10, 2016 10:59 AM, "Mullen, Mary via Histonet" 
 wrote:
 

 Hello all,



I work in a small, low volume community hospital and was recently asked by a 
coworker why we do not just run both our cytology and histology slides on the 
same automated stainer (with their respective protocols).



What I am wanting to know is if there is anyone currently running both staining 
protocols on a single automated stainer using common alcohols/xylenes/water? 
What are the pros/cons? Has there been any cross-contamination issues?



We only run non-gyn cytology, all gyn cytology is sent out.







Thanks,



Mary K. Mullen, HTL(ASCP)CM
Histotechnologist
UPMC Northwest
Seneca, PA

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Re: [Histonet] picric acid

[Rene J Buesa via Histonet]

Picric acid is an expensive reagent useful in many histology procedures.The 
advise you received of adding water is a good one.Humid picric acid will not 
explode at all. Why waste a good reagent?Keep humid, you will eventually used 
it.René 

On Thursday, May 5, 2016 3:24 PM, Mca Werdler via Histonet 
 wrote:
 

 Dear histonetters,

Since a few months, i started working in a histology lab, run only by me (
coworkers are not specialized in histology). There has not worked here a
person at histology for about 2 years.

After many new protocols, i decided to clear out some chemicals.
Now i found around 1 KG of DRY picric acid. I informed my coworkers about
this, and they said just to dissolve everything in water.

What do you guys think is the best way for handeling with this explosive
chemical? Thank you all in advance!

Maarten
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Re: [Histonet] PAS Stain

[Rene J Buesa via Histonet]

As I see it, there are 3 main objections about using human saliva as an amylase 
source.In order of importance they are:1- you will never know the actual 
concentration of the amylase and this will produce reproducibility problems.2- 
along with the saliva you will introduce bacteria that may end being stained 
and can cause misdiagnoses.3- it is absolutely disgusting.René 

On Thursday, May 5, 2016 2:26 PM, Bob Richmond via Histonet 
 wrote:
 

 Amylase (diastase) for the PAS stain queries:

Whatever happened to spitting on the slide (30 min at room temperature)?
John Kiernan advises "thinking of lemons and drooling into a small beaker"
though I'd advise chewing on a rubber band for a few seconds.

He notes that alpha amylase is preferred. I'd go with the cheapest one in
the Sigma-Aldrich catalog. Room temperature is usual, but I note that Sigma
offers a heat-stable alpha amylase.

Bob Richmond
Samurai Pathologist
Maryville TN
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Re: [Histonet] replacing a stainer and a coverslipper question

[Rene J Buesa via Histonet]

Productivity and quality sake, Sakura film coverslipper has no match. If you 
use Sakura tape and the xylene dispenser is properly calibrated, storage is not 
an issue.Sakura stainer was also what I used at my lab and I highly recommend 
both.René 

On Wednesday, May 4, 2016 3:19 PM, Jenn via Histonet 
 wrote:
 

 Dear Histonetters,
We are going to replace our automated stainer (LeicaXL) and coverslipper within 
the next few months.  I would really appreciate any suggestions that people may 
have about which stainer/coverslipper units they use/used and are happy with.

I am also curious about tape vs glass coverslippers.  We have always had glass 
here, but I know that there are people who love their tape units.  I would be 
willing to look at a tape unit, but I am concerned about long term storage 
issues.  I have seen a few postings lately about the tape coming off of the 
slides and that scares me!  I would appreciate any advice on what brands work 
well and which ones to avoid, if you are willing to share...

If you have any insight that you would like to share, you can post it or e-mail 
me directly, especially if it is about which units/tapes to avoid.  I don't 
want to vendor bash, so please feel free to email me directly  at 
jennifer.john...@genzyme.com
Thanks!
Jenn

Jennifer Johnson, B.S., HTL (ASCP)

Scientist
Department of Pathology
TEL.: 508-271-3610  -  Fax.: 508-872-9080
5 The Mountain Road, Framingham, MA 01701-9322
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Re: [Histonet] Automated IHC instrument

[Rene J Buesa via Histonet]

I tested those you mention and leased/used the one from DAKO and "never looked 
back".René 

On Wednesday, May 4, 2016 12:13 PM, "Murphy, Valerie via Histonet" 
 wrote:
 

 Hello Histonetters,
Our tissue core is interested in purchasing an IHC instrument. It would  be 
used for the more routine assays such as ER/PR, Her2, Ki67 etc.
Can anyone make a recommendation? Leica, Ventana, Dako?

Thank you,

Valerie Ratliff  B.Sc HTL(ASCP)
Research Assistant
Department of Oncology
Karmanos Cancer Institute
4100 John R
Detroit, MI 48201

Telephone: (313) 576 8282
Fax: (313) 576 8306
E-mail: 
murp...@karmanos.org

Better treatments. Better outcomes.


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Re: [Histonet] PAS/Decal Question

[Rene J Buesa via Histonet]

My impression is that your problem is during the decalcification step. It 
cannot be hurried and has to be in EDTA at pH 7All reagents have to be prepared 
in pH7 phosphate buffer.The inconsistency resides in the fact that not all core 
Bx are the same regarding thickness, tissue condition or size.Besides you are 
hurrying too much. As yourself (and your pathologists) the following question: 
what good you take out of your protocol if the "failure" rate is as big as you 
describe?Change to EDTA and process more time.René 

On Tuesday, May 3, 2016 1:01 PM, "Marcum, Pamela A via Histonet" 
 wrote:
 

 We are still having issues with our PAS stain on decaled bone marrows.  The 
Pathologists in HemePath are seeing what they refer to as smudginess in cells 
on some areas of the completed PAS slides.  We have looked at everything and 
cannot find where the issue is coming from at this point.  We have done manual 
staining for PAS, automated on the Leica stainer and on the Dako Artisan.  All 
methods show the same result for some slides.  We can go for several days to a 
week or more with no problem and then suddenly it is back and we have changed 
nothing in the way we do the processing, embedding, sectioning, 
deparaffinization and coverslipping.  We do as many as 38 bone marrow cores a 
night or as few as 8 and can find no correlation in the number we have to deal 
with for a given period.  All bone marrows drawn today must be completed by 8AM 
tomorrow morning.

Fixation after pulling the bone marrows is a minimum of 2 hours in AZF with a 
maximum of 7 hours +/-.

Grossed and placed in cassettes for 15 minute rinse in running DI Water

Decal currently in the Milestone Decal Unit for 45 minutes in Immunocal at 50C

Rinsed in running DI water for 15 minutes

Placed in 10% NBF and processed on a 4 hour program with a delay of 4 hours 
minimum to come off at 4:45AM.

If anyone knows of any literature on decal effects on PAS staining in bone 
marrows please contact me.  This has been going on for months and no matter 
what we do manual staining, Leica adaptation for automated or Dako it is not 
helping.  Dako has been great with sending in technical experts repeatedly and 
we cannot get this corrected.

Thanks,
Pam

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Re: [Histonet] Fwd:

[Rene J Buesa via Histonet]

As I see it, the best solution is "1"Even more: if the piece of tissue is large 
enough,→cut 1 section and stain → select at least 2 (+) areas→ divide the block 
into 2 blocks each containing one of those 2 areas and by doing so you would 
have duplicated the number of possible (+) sections.René 

On Monday, May 2, 2016 3:09 PM, Linda Margraf via Histonet 
 wrote:
 

 

> From: Cindy Bulmer 
> Date: May 2, 2016 at 12:15:22 PM CDT
> To: histonet-ow...@lists.utsouthwestern.edu
> 
> Hello Histoland,
>  
> I need some advice, I have a PT block that is positive with Spirochetes.
> What would be the best way to use this block as a positive control?
>  
> 1)  Cut (serial sections, stain the last slide for bugs) and oven time (60) 
> for 1 hr.
>      then put slides in refrigerator for future use.
> 2)  Cut (serial sections, stain the last slide for bugs) NO oven time  and 
> put slides
>      directly in refrigerator for future use.
> 3)  Cut "fresh" every time they order the Ab.
>  
> Thank you,
> Cindy
> Cynthia Bulmer HT(ASCP),QIHC
> IHC Supervisor, CTPL
> Waco, TX
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Re: [Histonet] No More Blog Posts -- Over and Out!

[Rene J Buesa via Histonet]

Thank you VERY MUCH!René 

On Monday, May 2, 2016 1:54 PM, Lester Raff MD via Histonet 
 wrote:
 

 To My Lab Colleagues:

As my intent has never been to sow discontent or rancor, I think it is for the 
best if I no longer post links to my blog, lab related or otherwise.  Of course 
the blogs go on, and if anyone is interested in being added to my mailing list 
for future notifications, just drop me a line at 
les.r...@post.com  The mailing list is never used for 
any purpose other than announcing a new blog post. Be sure to let me know you 
are from the Histonet list!

I will continue to participate in any histology/pathologist discussions here as 
I have for many years.

Cheers,

Lester J. Raff, MD MBA
UroPartners
Medical Director Of Laboratory
2225 Enterprise Dr. Suite 2511
Westchester, Il 60154
Tel: 708-486-0076
Fax: 708-492-0203

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Re: [Histonet] Medical/health related post

[Rene J Buesa via Histonet]

Bryan:1- who gave permission to Dr. Raff?2- how we know the permission was 
given?3- what percentage of HistoNet member gave the permission?4- why Dr. Raff 
is so stubborn to keep posting what ever he chooses in spite of the rejection 
of probably more members than those who "gave him permission"?5- you write 
about some "rude" comments, how about Dr. Raff's arrogant disregard for the 
opinion of others?René 

On Thursday, April 28, 2016 4:06 PM, Bryan Llewellyn via Histonet 
 wrote:
 

 I think it is rather unfair to accuse Dr. Raff of harassment or misusing 
this forum. I recall that when he first posted references to his blog he 
asked for the members' permission to do so, and he was given that 
permission. Rather than abusing the privilege of posting to Histonet, I 
think he showed distinct courtesy for asking before he did so, and 
remarkable restraint in not responding to the, sometimes rude, 
responses. If members have now changed their minds about giving 
permission to him to past references to his blog, then let that 
permission be withdrawn with the same courtesy as he showed when first 
asking for it.

Incidentally, it takes me about 0.5 seconds to click delete on his 
posts. I do not read them since I am a Canadian and have no real 
interest in US health care experiences or problems.

Bryan Llewellyn



Tyra Connor via Histonet wrote:
> I have been on this list for some time now, and I have seen Dr. Raff 
> contribute very useful information to many posed questions. Recently this 
> situation has shifted, with more posts concerning his blog than actual 
> helpful responses. This is not the second or even third time this situation 
> has been addressed in this forum. It's sad because I would not want to lose 
> his voice in these discussions, but these days whenever I see any post from 
> him at all, I immediately delete it. I feel at this point that his refusal to 
> stop is a small form of harassment, like a telemarketer that refuses to stop 
> calling even though you request that they do so.
>
> Be full of wonder, be wonderful!!
>
>> On Apr 28, 2016, at 10:48 AM, WILLIAM DESALVO via Histonet 
>>  wrote:
>>
>> Again, why the non-Histo post. Take this to another source. I do not 
>> understand why Dr. Raff has not been removed from this list serve. This is a 
>> valuable site for histotechnolgy related issues, please let us keep it that 
>> way.
>>
>> Sent from my Windows Phone
>> 
>> From: Lester Raff MD via Histonet
>> Sent: ‎4/‎28/‎2016 6:59 AM
>> To: 
>> 'histonet@lists.utsouthwestern.edu'
>> Subject: [Histonet] Medical/health related post
>>
>> http://www.chicagonow.com/downsize-maybe/2016/04/running-mates-and-other-mates-fighting-trump-fighting-cancer/
>>
>> Lester J. Raff, MD MBA
>> UroPartners
>> Medical Director Of Laboratory
>> 2225 Enterprise Dr. Suite 2511
>> Westchester, Il 60154
>> Tel: 708-486-0076
>> Fax: 708-492-0203
>>
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Re: [Histonet] Using same DAB for multiple slide racks

[Rene J Buesa via Histonet]

This is not a good practice and can lead to inconsistent results.Always use 
freshly prepared DAB sol. to finally be able to see the end product of the 
costly and important IHC procedure, it is worth it.René 

On Monday, April 25, 2016 6:27 PM, Andrea Calhoun via Histonet 
 wrote:
 

 Hi Listers!

Has anyone ever tried re-using DAB? Not like week or day old DAB, but using the 
same DAB solution for more than one rack of slides? Anyone know the length of 
time DAB is activated for before becoming inactivated, or quenched?  Safe to 
do, if more H202 is added?

After antibody validation, we typically know how long to stain with DAB so we 
can stain a huge batch (24slides/rack, normally 2-3 racks) each in their own 
150mls of Dab solution. But the DAB tablets we get are expensive and we are 
seeing an increase in the amount of IHC runs we're doing- so the whole thing is 
becoming expensive.  If it is viable to use the same 150mls of DAB for 2-3 
racks of slides, that would greatly decrease the amount we're using and cost.

If anyone has ever tried this, or does this routinely- please let me know!

Andrea Calhoun

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Re: [Histonet] copper stain

[Rene J Buesa via Histonet]

Timm's silver stain always!René 

On Wednesday, April 20, 2016 12:51 PM, Gudrun Lang via Histonet 
 wrote:
 

 Hi all!

Which stain would you prefer to demonstrate copper? Rhodanin or Victoria blue?

 

Thanks in advance

Gudrun

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Re: [Histonet] Blog Post Not lab related

[Rene J Buesa via Histonet]

Amen!!!But you have to concede that Lester is a very  persistent, almost 
obstinate individual, probably used to impose his will and this postings are 
just an example of it: he likes his blog and tries to impose it to everyone. 
Evidently he has all the time in the world and just does not know what to do 
with it and enjoys sharing his "witty" side. René 

On Wednesday, April 13, 2016 3:24 PM, Caroline Miller via Histonet 
 wrote:
 

 Lester, I do believe this is the second 'non lab related' blog post this
week. As we have spoken about before I do not think histonet is an
appropriate place for your blog posts. I, personally, was totally OK with
you co-posting with other lab related topics. However you are now pushing
it and I respectfully ask that you refrain from sending out your blog posts
to the whole list, let people sign up if they are interested.

The issue is that this is a histology related list, if everyone posted
non-lab stuff here it would soon be chaos!

Respectfully yours,
mills

On Wed, Apr 13, 2016 at 11:38 AM, Lester Raff MD via Histonet <
histonet@lists.utsouthwestern.edu> wrote:

> Just some philosophy towards the end of a long day.
>
>
> http://www.chicagonow.com/downsize-maybe/2016/04/kitten-power-can-get-things-done/
>
>
>
> Just a reminder-I try to limit my blog invitations here. If you enjoy the
> blog, remember to subscribe (no charge, no spam) on the ChicagoNow blog
> site.
>
> Thanks for your readership.
>
> Lester J. Raff, MD MBA
> UroPartners
> Medical Director Of Laboratory
> 2225 Enterprise Dr. Suite 2511
> Westchester, Il 60154
> Tel: 708-486-0076
> Fax: 708-492-0203
>
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>



-- 
Caroline Miller (mills)
Director of Histology
3Scan.com
415 2187297
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Re: [Histonet] H automated stainers

[Rene J Buesa via Histonet]

Before deciding, ask for a "demo" from Sakura.René 

On Monday, April 11, 2016 11:02 AM, Lauren Marie Hegner via Histonet 
 wrote:
 

 Hello all,


Our lab is looking for a new automated H stainer and I was wondering if 
anyone out there has had any experience using Ventana's Symphony System, 
Hacker's HistoPro 3030, or Leica ST5010 Autostainer XL?


OR


If anyone has any suggestions as to the best one out there?


Thanks,

L


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Re: [Histonet] Porcessing FFPE tissue without alcohol??

[Rene J Buesa via Histonet]

Sudan Black reacts only with protein-combined fats.René 

On Saturday, March 26, 2016 11:20 AM, Joanna <jobalu...@gmail.com> wrote:
 

 How about Sudan Black stain? 

> On Mar 26, 2016, at 4:32 AM, Rene J Buesa via Histonet 
> <histonet@lists.utsouthwestern.edu> wrote:
> 
> The only problem I see is that the fat will be preserved, as you wrote, as a 
> black osmium oxidate but you will not be able to use any "standard" fat 
> stain; otherwise it will work.René 
> 
>    On Friday, March 25, 2016 2:41 PM, "Hobbs, Carl via Histonet" 
><histonet@lists.utsouthwestern.edu> wrote:
> 
> 
> 
> 
> Fix the tissue in Formalin, wash well in dw, then place very small pieces 
> into Osmium tetroxide solution ( std soln for TEM post-fixation)
> Processing to Pwax as usual.
> Basically, you will see lipids as black ( oxidised osmium)
> That's the only way to demonstrate solvent- soluble lipids, using Pwax 
> processing.
> Sure, there are caveats but, in the main...it will be Ok, imho.
> I invite comments as I may be doing exactly this very soon, to count 
> myelinated nerve fibres in a sciatic nerve.
> 
> 
> 
>  
> 
> Carl Hobbs FIBMS 
> Histology and Imaging Manager 
> Wolfson CARD 
> Guys Campus, London Bridge  
> Kings College London 
> London 
> SE1 1UL 
>  
> 020 7848 6813    
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Re: [Histonet] Porcessing FFPE tissue without alcohol??

[Rene J Buesa via Histonet]

The only problem I see is that the fat will be preserved, as you wrote, as a 
black osmium oxidate but you will not be able to use any "standard" fat stain; 
otherwise it will work.René 

On Friday, March 25, 2016 2:41 PM, "Hobbs, Carl via Histonet" 
 wrote:
 

 

Fix the tissue in Formalin, wash well in dw, then place very small pieces into 
Osmium tetroxide solution ( std soln for TEM post-fixation)
Processing to Pwax as usual.
Basically, you will see lipids as black ( oxidised osmium)
That's the only way to demonstrate solvent- soluble lipids, using Pwax 
processing.
Sure, there are caveats but, in the main...it will be Ok, imho.
I invite comments as I may be doing exactly this very soon, to count myelinated 
nerve fibres in a sciatic nerve.



  
 
Carl Hobbs FIBMS 
Histology and Imaging Manager 
Wolfson CARD 
Guys Campus, London Bridge  
Kings College London 
London 
SE1 1UL 
  
020 7848 6813    
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Re: [Histonet] Porcessing FFPE tissue without alcohol??

[Rene J Buesa via Histonet]

To embed the tissues with paraffin you HAVE TO dehydrate the tissue. This is 
usually done with either ethanol of 2-propanol but essentially all dehydrants 
will remove fat so you are right, the way to go is going frozen sections.René 

On Thursday, March 24, 2016 2:24 PM, "Dessasau III, Evan via Histonet" 
 wrote:
 

 Hi Histonet,  is there a way to process tissue for paraffin embedding without 
using alcohol?                        One of the labs that send their 
processing to us will be doing a study examining  fat.  I told them if they 
want to look at the fat they will have to cut frozen sections but I'm being ask 
again about processing without the alcohol.  So I said I would ask around.
Please let me know what you think.
Thank you for your help!
E-van



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Re: [Histonet] Whale skin

[Rene J Buesa via Histonet]

How did you manage to deal with the about 0.5 m of blubber?Was it the skin of a 
new born whale? I just to not understand, but you have to completely eliminate 
all the fat and increase your processing protocol (infiltration specially) to 
have some chance of getting any relatively "decent" section.René 

On Monday, March 7, 2016 11:23 AM, Kathleen Jones via Histonet 
 wrote:
 

 Hello, 

I have to cut FFPE blocks of whale skin for in IHC study. I am having
difficulty with both cutting and floating. Any advice would be much
appreciated!

Thank you!





Kathleen Jones
Research Technologist
Pathology/Microbiology
AVC - UPEI


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Re: [Histonet] question - Allergy to histological solvents?

[Rene J Buesa via Histonet]

You can dewax absolutely safely using a 2% dishawasher soap solution at 90ºC 
(twice) as washing in water.You can "dehydrate" stained stains by placing the 
slides in an oven at 60ºC, also absolutely safely for the stained section.Under 
separate cover I am sending  articles on this subject.René 

On Saturday, February 20, 2016 3:05 PM, daniel blackburn via Histonet 
 wrote:
 

 Hello

After decades of using standard organic solvents for paraffin- section 
histology, I find that I've become highly allergic to the fumes.  These include 
commercial preps like Citrosolv and Hemo-De, as well as xylene and toluene, 
which cause (in me) dizziness and a pounding headache.  This problem has made 
staining and coverslipping very difficult, even with  hood.  Can anyone 
recommend alternative solutions for (a) dewaxing prior to staining and (b) 
coverslipping?  thanks very much!

Daniel Blackburn
Biology, Trinity College (Hartford CT)

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