Re: [Histonet] Inquiry on Tissue Softening for Microtomy

[Tony Henwood via Histonet]

My preference is cold water or Mollifex on faced blocks.

Ammonia is definitely obnoxious (reminds me of wet nappies )


Regards,



Tony Henwood MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) (Retired)

Principal Scientist, the Children’s Hospital at Westmead (Retired)

Adjunct Fellow, School of Medicine, University of Western Sydney.


From: Jay Lundgren via Histonet 
Sent: 13 May 2024 08:37
To: John Kiernan 
Cc: histonet@lists.utsouthwestern.edu ; 
IGNACIO GONZÁLEZ MASSONI 
Subject: Re: [Histonet] Inquiry on Tissue Softening for Microtomy

Pleasant is not a mission parameter.
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Re: [Histonet] Histonet Digest, Vol 245, Issue 20

[Tony Henwood via Histonet]

A few years ago there was an article on the Block - "What is Denatured Alcohol 
and What are the Implications For Histopathology?" that might be useful. The 
URL is

https://www.nsh.org/blogs/tony-henwood/2020/02/11/what-is-denatured-alcohol-and-what-are-the-implica?_gl=1*1mgiiv6*_ga*MTkyMzY5MTQ4Mi4xNzExNDA1ODQ2*_ga_7J0VYE6519*MTcxNDU0NzgyNy4xMC4xLjE3MTQ1NDc4NzcuMC4xLjEzNzYzNTQ4NjQ.&_ga=2.47649445.329273616.1714547828-1923691482.1711405846



Regards,



Tony Henwood MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) (Retired)

Principal Scientist, the Children’s Hospital at Westmead (Retired)

Adjunct Fellow, School of Medicine, University of Western Sydney.


From: John O’Brien via Histonet 
Sent: 01 May 2024 01:49
To: histonet@lists.utsouthwestern.edu 
Subject: Re: [Histonet] Histonet Digest, Vol 245, Issue 20

Ethyl Alcohol,
Reagent 100% alcohol is what is normally used in Pathology processing and 
slide staining
IMEB Inc offer all grades of Reagent alcohol, pure ethyl is a controlled 
alcohol regulated and taxed by government
Regards
John

> On Apr 30, 2024, at 10:18 AM, histonet-requ...@lists.utsouthwestern.edu wrote:
>
> 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. Alcohol (Naira Margaryan)
>
>
> --
>
> Message: 1
> Date: Mon, 29 Apr 2024 18:49:04 -0500
> From: Naira Margaryan 
> To: Histonet 
> Subject: [Histonet] Alcohol
> Message-ID:
>
> Content-Type: text/plain; charset="UTF-8"
>
> Hello,
>
> What type of ALCOHOL ETHYL you?re using in your histology lab.?
>
> We are thinking to get 20-30-gal drum.
>
> Could you please help me with that?
>
>
>
> Your suggestion is appreciated,
>
> Naira
>
>
> --
>
> Subject: Digest Footer
>
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> --
>
> End of Histonet Digest, Vol 245, Issue 20
> *


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Re: [Histonet] Modified Davidson solution

[Tony Henwood via Histonet]

I agree.

A great book.


Regards,



Tony Henwood MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) (Retired)

Principal Scientist, the Children’s Hospital at Westmead (Retired)

Adjunct Fellow, School of Medicine, University of Western Sydney.


From: John Kiernan via Histonet 
Sent: 06 March 2024 18:32
To: Histonet ; Naira Margaryan 
; Davoli, Katherine A 
Subject: Re: [Histonet] Modified Davidson solution

The Davidson/Hartmann fixative is just another alcoholic formaldehyde mixture 
acidified with acetic acid. Like all others of that ilk it was intended for 
making up in the lab soon before using. Storage causes slow deterioration. The 
ethanol slowly gets esterified by the acetic acid, making ethyl acetate, which 
has no fixative properties. This change is evident from the fruity smell of the 
ester, which becomes evident after a week or two. All this has been in the 
textbooks for more than 50 years.
Why doesn't every lab spend perhaps $200 on a couple of books?  For obvious 
reasons I recommend my 5th edition (ISBN 9781907904325; $82.50,from the 
publisher, which is $30 less than Amazon's price), but there are other 
histotechnology books that are very good.  A book may cost much less than a big 
bottle of a deteriorating mixture of questionable value, and you and your staff 
could learn a lot by reading.
John Kiernan
= = =

From: Davoli, Katherine A via Histonet 
Sent: March 5, 2024 4:40 PM
To: Histonet ; Naira Margaryan 

Subject: Re: [Histonet] Modified Davidson solution

Hi Naira, I get Hartmann's from Electron Microscopy Sciences
Cat# 64133-10 (for 1L, but they have larger sizes I believe)


Katherine Davoli, HTL(ASCP)cm(they/them/theirs)

Lab Manager, Tissue Culture and Histology Core Facilities

U. Pitt Dept of Ophthalmology

7.373 UPMC Mercy Pavilion

1622 Locust St., Pittsburgh PA 15219

(412) 624-8508   this number cannot receive texts


From: Naira Margaryan via Histonet 
Sent: Tuesday, March 5, 2024 1:27 PM
To: Histonet 
Subject: [Histonet] Modified Davidson solution

Hello!

What is the best company to buy a ready to use Modified Davidson solution?

Thanks in advance,
Naira
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From: Davoli, Katherine A via Histonet 
Sent: March 5, 2024 4:40 PM
To: Histonet ; Naira Margaryan 

Subject: Re: [Histonet] Modified Davidson solution

Hi Naira, I get Hartmann's from Electron Microscopy Sciences
Cat# 64133-10 (for 1L, but they have larger sizes I believe)


Katherine Davoli, HTL(ASCP)cm(they/them/theirs)

Lab Manager, Tissue Culture and Histology Core Facilities

U. Pitt Dept of Ophthalmology

7.373 UPMC Mercy Pavilion

1622 Locust St., Pittsburgh PA 15219

(412) 624-8508   this number cannot receive texts


From: Naira Margaryan via Histonet 
Sent: Tuesday, March 5, 2024 1:27 PM
To: Histonet 
Subject: [Histonet] Modified Davidson solution

Hello!

What is the best company to buy a ready to use Modified Davidson solution?

Thanks in advance,
Naira
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Re: [Histonet] Modified Davidson

[Tony Henwood via Histonet]

There are several formulations (some are probably typos), but this seems to be 
one commonly cited.

Modified Davidson's Fixative:

(Latendresse, J. R., Warbrittion, A. R., Jonassen, H., & Creasy, D. M. (2002). 
Fixation of testes and eyes using a modified Davidson's fluid: comparison with 
Bouin's fluid and conventional Davidson's fluid. Toxicologic pathology, 30(4), 
524-533.)

37–40% formaldehyde30ml
Absolute ethanol 15ml
Glacial acetic acid5ml
Distilled Water 50ml

Regards,

Tony Henwood MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) (Retired)
Principal Scientist, the Children’s Hospital at Westmead (Retired)
Adjunct Fellow, School of Medicine, University of Western Sydney.


From: Naira Margaryan via Histonet 
Sent: Friday, February 9, 2024 8:25:47 AM
To: Histonet 
Subject: [Histonet] Modified Davidson

Hello,

Could you please share your best recipe for the Modified Davidson?

Thanks in advance,
Naira
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Re: [Histonet] Processing artifact - delayed start

[Tony Henwood via Histonet]

I would check the level of the formalin after it has been pumped into the 
retort.

I will wait till the pics are posted

Regards,

Tony Henwood MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) (Retired)
Principal Scientist, the Children’s Hospital at Westmead (Retired)
Adjunct Fellow, School of Medicine, University of Western Sydney.


From: Verizon wireless via Histonet 
Sent: Saturday, February 3, 2024 2:36:42 PM
To: histonet@lists.utsouthwestern.edu 
Subject: [Histonet] Processing artifact - delayed start

Dear Histonet Members,

We have terrible processing artifact if tissue sits in the formalin-filled 
retort (at ambient temperature) for too long (more than 10-12 hours) before a 
delayed process starts. The longer the wait, the worse it looks. We have Tissue 
Tek VIP 5 processors, and we process luminal gastrointestinal biopsies 
exclusively. I've attached some photomicrographs of problem cases on the 
Histonet Images website (with the same topic title).
This artifact typically affects a few specimens per day (~2% or less), even 
though everything is done on the same processor; it may affect all tissue 
portions in a cassette or only some of the tissue in that cassette. Some tissue 
portions may only have the artifact on one half with the other half looking 
perfectly fine. The techs sometimes note the tissue feeling "crunchy" at the 
time of embedding and / or at the time of microtomy. These tissues tend to 
suffer greater chatter artifact and have trouble sticking to the slides. The 
sections look just as bad on recuts as the originals. Re-processing does not 
seem to help at all.
If the cassettes sit in formalin in a container outside of the processor for 
days before the processor is loaded (with subsequent immediate start), things 
look perfectly fine. When we have staff around to start the processor 
immediately upon loading cassettes and empty immediately upon completion, the 
tissue looks perfectly fine.

Our current processing program is as follows:

1. 10% Formalin, 30 minutes, ambient temp, p/v on
2. 10% Formalin, 30 minutes, ambient temp, p/v on
3. 65% Alcohol, 10 minutes, ambient temp, p/v on4. 80% Alcohol, 10 minutes, 
ambient temp, p/v on5. 95% Alcohol, 7 minutes, ambient temp, p/v on6. 95% 
Alcohol, 7 minutes, ambient temp, p/v on 7. 100% Alcohol, 10 minutes, ambient 
temp, p/v on
8. 100% Alcohol, 10 minutes, ambient temp, p/v on
9. Xylene, 15 minutes, ambient temp, p/v on
10. Xylene, 15 minutes, ambient temp, p/v on
11. Paraffin, 10 minutes, 60 degrees C, p/v on12. Paraffin, 10 minutes, 60 
degrees C, p/v on13. Paraffin, 15 minutes, 60 degrees C, p/v on14. Paraffin, 15 
minutes, 60 degrees C, p/v on

Obviously there are times we absolutely need to use delayed start. I would 
greatly appreciate guidance, and I'll be happy to provide any other details 
that might be useful.
Sincerely,
Brian Quigley MDLaboratory Director of a GI Pathology Laboratory

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Re: [Histonet] Faded H tissue section

[Tony Henwood via Histonet]

I thought he may have reported this in Histologic.
This trick certainly works. You can notice the nuclear clarity on GIT and to a 
lesser extent skin biopsies stained with PAS.

Regards,

Tony Henwood MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) (Retired)
Principal Scientist, the Children’s Hospital at Westmead (Retired)
Adjunct Fellow, School of Medicine, University of Western Sydney.

From: Akemi Allison
Sent: Friday, 10 November 2023 1:30 PM
To: jayalakshmy p.s
Cc: Tony Henwood; 
histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] Faded H tissue section

Hi Tony:
I actually gave that tip to Lee back in 1979 when he came to OHSU to give a 
seminar to the Oregon Histology Society.  I found out that when I was doing GMA 
and the pathologists didn’t like Gills Hematoxylin. They loved my PAS on GMA so 
I tried using 1% Periodic Acid before using Harris Hematoxylin for H’s on GMA 
and it turned out beautifully. Guess he shared my tip with the rest of our 
society. He and I became closer se friends and we shared several tips, 
including his Movat’s tips which he didn’t publish, but I shared them in 
Frieda’s 4th Ed.

Best,
Akemi Allison-Tacha, BS, HT/HTL (ASCP)

Sent from my iPhone

> On Nov 9, 2023, at 6:11 PM, jayalakshmy p.s via Histonet 
>  wrote:
>
> Thanks, I'll check it out.
>
>> On Fri, Nov 10, 2023, 6:58 AM Tony Henwood  wrote:
>>
>> You could treat the decoverslipped section in 1% periodic acid (same as
>> used in the PAS technique)  for 30 minutes prior to H staining. This
>> might improve the H staining.
>>
>>
>>
>> I believe Lee Luna suggested this but for the life of me, I can’t find the
>> reference!
>>
>>
>>
>> Regards,
>>
>>
>>
>> Tony Henwood MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) (Retired)
>>
>> Principal Scientist, the Children’s Hospital at Westmead (Retired)
>>
>> Adjunct Fellow, School of Medicine, University of Western Sydney.
>>
>>
>>
>> *From: *jayalakshmy p.s via Histonet 
>> *Sent: *Friday, 10 November 2023 3:43 AM
>> *To: *histonet@lists.utsouthwestern.edu
>> *Subject: *[Histonet] Faded H tissue section
>>
>>
>>
>> Hello,
>> I would like to know how to effectively restain a faded H tissue section.
>> The color becomes dull when re stained. Somebody please advise.
>> Prof. Jayalakshmy. P S
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>>
>>
>>
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Re: [Histonet] Faded H tissue section

[Tony Henwood via Histonet]

You could treat the decoverslipped section in 1% periodic acid (same as used in 
the PAS technique)  for 30 minutes prior to H staining. This might improve 
the H staining.

I believe Lee Luna suggested this but for the life of me, I can’t find the 
reference!

Regards,

Tony Henwood MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) (Retired)
Principal Scientist, the Children’s Hospital at Westmead (Retired)
Adjunct Fellow, School of Medicine, University of Western Sydney.

From: jayalakshmy p.s via Histonet
Sent: Friday, 10 November 2023 3:43 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Faded H tissue section

Hello,
I would like to know how to effectively restain a faded H tissue section.
The color becomes dull when re stained. Somebody please advise.
Prof. Jayalakshmy. P S
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Re: [Histonet] Formalin pH

[Tony Henwood via Histonet]

Hew-Shue (1991) has described a useful pH indicator for working neutral 
buffered formalin solutions. Bromocresol purple, when added to formalin 
solutions, serves as an indicator of pH as well as, from a safety aspect, 
labelling the solution as formalin making redundant the dangerous “smell” test. 
At an acidic pH (5.2) the intense indicator colour is yellow whereas at pH 6.8, 
the colour is purple. A saturated solution of the dye is prepared, and 2 to 4 
drops are added to 10 litres of neutral buffered formalin. Other advantages are 
that the fixative is more readily distinguished from other colourless solutions 
such as saline, thus preventing accidental misuse, and formalin spills are more 
easily recognised.

Hew-Shue N (1991) “Bromcresol Purple as a Colored Marker and pH Indicator for 
Ten Percent Neutral Buffered Forrnalin” J Histotechnol 14(4):257-260.


Regards,

Tony Henwood MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) (Retired)
Principal Scientist, the Children’s Hospital at Westmead (Retired)
Adjunct Fellow, School of Medicine, University of Western Sydney.

From: Bacon, Charles via Histonet
Sent: Thursday, 14 September 2023 6:42 AM
To: e...@pigs.ag
Cc: HistoNet
Subject: Re: [Histonet] Formalin pH

We pull the COA from the vendors website. Some are better than others but the 
are usually listed by lot number. I keep them in a network shared folder so 
they can be pulled by anyone that is asked for the evidence during inspection.

Sent from my iPhone

> On Sep 13, 2023, at 8:22 AM, "e...@pigs.ag"  wrote:
>
> Sooner or later the people have got to rise up
> and tell those brain-dead paper-shuffling regulators
> to just stuff it.  Their mindless interference is
> just wasting time and interfering with people's
> ability to keep their own garden.
>
> -Original Message-
> From: Cooper, Brian via Histonet
> Reply-To: Cooper, Brian
> To: Paula Sicurello, HistoNet
> Subject: Re: [Histonet] Formalin pH
> Date: Today 10:19 AM
>
> Ask your vendor to send a certificate of analysis that includes the pH. 
> That's what we did and now they come with each lot. We do have to bug them 
> for them occasionally.
>
> I'm betting your vendor has at least heard about this from other customers so 
> it should be on their radar at the very least.
>
>
> Thanks,
>
>
>
> Brian D. Cooper, HT (ASCP)CMQIHCCM| Histology Supervisor
>
> Department of Pathology and Laboratory Medicine
>
> Children's Hospital Los Angeles
>
> 4650 Sunset Blvd MS#43- Los Angeles, CA 90027
>
> Ph: 323.361.3357
>
> bcoo...@chla.usc.edu
>
> 
> From: Paula Sicurello via Histonet 
> Sent: Tuesday, September 12, 2023 7:12:05 PM
> To: HistoNet 
> Subject: [Histonet] Formalin pH (EXTERNAL EMAIL)
>
> CAUTION: BE CAREFUL WITH THIS MESSAGE*
> This email came from outside CHLA. Do not open attachments, click on links, 
> or respond unless you expected this message and recognize the email address: 
> histonet-boun...@lists.utsouthwestern.edu.
>
> Hello Histoteckies,
> What are y'all doing regarding the CAP requirement to monitor the pH of 
> formalin?
> We buy tons and tons of the 5 gallon cubitainers and we are still debating 
> over how to check the pH.
>
> Looking forward to your replies.
> Toodles!
> Sincerely,
>
> Paula Sicurello
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Re: [Histonet] shelf life of working antibody solutions in IHC

[Tony Henwood via Histonet]

In Australia, we have evolved from Prescriptive accreditation (following the 
standards) to Risk-based accreditation where it is more important to assess the 
risk of a laboratory process rather than only meeting a list of rules, since 
following a list of rules is often not in the patient’s nor in the community’s 
interest.

Years ago, I battled with our accreditation organization over the risk involved 
in fridge temperature recording, presenting evidence that continuous digital 
monitoring (with an alarm system) was less risky than recording max/min 
temperatures daily (eg what happens when the lab is not manned during Christmas 
holidays and weekends?).

The excessive cost involved in automatically disposing of expired antibodies is 
wasteful. Continuous monitoring as well as preventative practices such as 
preservatives (azide, procillin, etc) and pH monitoring with phenol red greatly 
reduces the risk.

As presented in the paper, many working dilutions survive without contamination 
for many years. For example, factor 8 (21 years), factor 13a (19 years) and 
epithelial membrane antigen (17 years). There were no failed verifications and 
to date, the average life after expiration was 6 years; eight antibodies 
exceeded 6 years. All the other antibodies would probably have similar survival 
times but were exhausted before reaching these times.

Regards,

Tony Henwood MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) (Retired)
Principal Scientist, the Children’s Hospital at Westmead (Retired)
Adjunct Fellow, School of Medicine, University of Western Sydney.

From: Gudrun Lang
Sent: Sunday, 2 July 2023 7:26 PM
To: 'Tony Henwood'
Subject: AW: [Histonet] shelf life of working antibody solutions in IHC

Dear Tony,
thank you for the interesting article. It reflects my personal experience. 
Nevertheless accreditation rules overrule the experience and in an diagnostic 
lab we have to discard the reagenses.

My question regards more the practical approach with working solutions.
What is your experience how long the working solutions are „good enough“ for 
using.

Thank you very much and kind regards
Gudrun

Von: Tony Henwood [mailto:afhenw...@outlook.com]
Gesendet: Samstag, 1. Juli 2023 23:41
An: Gudrun Lang
Betreff: RE: [Histonet] shelf life of working antibody solutions in IHC

Here it is

Regards,

Tony Henwood MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) (Retired)
Principal Scientist, the Children’s Hospital at Westmead (Retired)
Adjunct Fellow, School of Medicine, University of Western Sydney.

From: Gudrun Lang via Histonet
Sent: Sunday, 2 July 2023 4:59 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] shelf life of working antibody solutions in IHC

Hi all!

I have tried to find a general instruction for the shelf life of antibody
working solutions.

With automated IHC you usually fill the container with the working solution
and depending on the frequency of usage they stay on the instrument at
roomtemperature (or higher) or are put in the fridge again.

The working solutions are up to 10 ml and may last for months. The
antibody-diluent is from the same company of the instrument. The titers are
in a range from 1:10 to 1:3000. - so a very heterogen situation.



How do you handle this? Have you a general rule, when the solution has to be
discarded? Is it just a matter of positiv-controls?



Thanks in advance

Gudrun Lang

Biomedical scientist

Austria





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Re: [Histonet] shelf life of working antibody solutions in IHC

[Tony Henwood via Histonet]

Hi Gudrun,

This should be useful ( I will send a copy separately)

Henwood, A. F. (2023). Validation of nominally expired antibodies for 
immunohistochemistry. Biotechnic & Histochemistry, 98(2), 86-93.

Regards,

Tony Henwood MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) (Retired)
Principal Scientist, the Children’s Hospital at Westmead (Retired)
Adjunct Fellow, School of Medicine, University of Western Sydney.

From: Gudrun Lang via Histonet
Sent: Sunday, 2 July 2023 4:59 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] shelf life of working antibody solutions in IHC

Hi all!

I have tried to find a general instruction for the shelf life of antibody
working solutions.

With automated IHC you usually fill the container with the working solution
and depending on the frequency of usage they stay on the instrument at
roomtemperature (or higher) or are put in the fridge again.

The working solutions are up to 10 ml and may last for months. The
antibody-diluent is from the same company of the instrument. The titers are
in a range from 1:10 to 1:3000. - so a very heterogen situation.



How do you handle this? Have you a general rule, when the solution has to be
discarded? Is it just a matter of positiv-controls?



Thanks in advance

Gudrun Lang

Biomedical scientist

Austria





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Re: [Histonet] Get the Short Term or Long Term Lab Staffing Coverage You Need

[Tony Henwood via Histonet]

I agree

Regards,

Tony Henwood
Sydney, Australia

From: John Kiernan via Histonet
Sent: Saturday, 3 June 2023 11:09 AM
To: Melissa Owens; Jay 
Lundgren
Cc: Tom Walls
Subject: Re: [Histonet] Get the Short Term or Long Term Lab Staffing Coverage 
You Need

Hear, Hear! Let's see less advertising on Histonet.
John Kiernan
(London, Canada).
= = =

From: Jay Lundgren via Histonet 
Sent: June 2, 2023 1:15 PM
To: Melissa Owens 
Cc: Tom Walls 
Subject: Re: [Histonet] Get the Short Term or Long Term Lab Staffing Coverage 
You Need

As far as I know, Histonet doesn't really allow advertising for individuals
or agencies *looking* for jobs.  There are a very few, selective, agencies
that are allowed to post on Histonet  tastefully and infrequently, but they
are buying, not selling.  Or histotechs who let others know about open
positions in their organization.  That's cool.

I'm not a moderator, but this is supposed to be a collegial forum for
practical histopathology advice.  There are plenty of job search sites out
there already.

But if Histonet does start allowing this, then ***%%%$$$HEY SCRIPPS
INSTITUTE HIRE ME$$$***

***%%%$$$LONG OR SHORT CONTRACT!!!CHEAP$$$%%%***

 **%%%$$$BIG TIME IMUNNOHISTOCHEMISTRY$$$%%%***

 ***%%%$$$USAF HTL (ASCP) M.S. 30+ YRS!!!$$$%%%***

 Sincerely,

  Jay A. Lundgren, M.S., HTL (ASCP)

On Fri, Jun 2, 2023 at 10:50 AM 'Melissa Owens' via Histonet <
histonet@lists.utsouthwestern.edu> wrote:

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Re: [Histonet] Sudanblack B on FFPET

[Tony Henwood via Histonet]

I would also let the saturated solution stand for a few days. Like Oil Red O, 
it probably needs time to “mature”. I would also use a frozen section of skin 
as a control.

Regards,

Tony Henwood
Sydney, Australia

From: Bob Richmond via Histonet
Sent: Wednesday, 29 March 2023 4:51 AM
To: Histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] Sudanblack B on FFPET

>
> Gudrun Lang in Austria asks:
>

>>Has anyone experience with Sudanblack B on paraffin slides for staining
[lipofuscin]? A doctor wants the demonstration of the lipoid content of
foamy cells or granulocytes in lung. I've found protocols that have
incubation-times from 10 minutes to over-night. - I've made a saturated
Sudan black B-solution in 70% ethanol and tried it with10 min on liver
without real success.<<

The main thing you need to do is demonstrate that it isn't hemosiderin with
an iron stain (Perls prussian blue reaction), and perhaps also that it
isn't acid-fast. Lipofuscin can be identified an H & E staining, except for
these considerations.

Bob Richmond
Maryville, Tennessee
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