Re: [Histonet] Treponema tissue blocks

2024-10-01 Thread Tony Henwood via Histonet
Hi John,

I'm still around (though now retired). Histopeeps? I suppose I have been called 
worse but my thick formalinised skin (or xylene addled brain, which assumes I 
had one in the first place) probably skips over the "Histopeeps". Anyway, I 
often marvel and I am truly appreciative of the value of Histonet to my 
professional life. Many posts have caused my (microtome) cogs turning.

I can communicate with thousands across the world in real time or wait 12 hours 
till everyone, not in our time zone, wakes up! Well Australia is a heck of a 
long way away.

Many seeds for my publications were sown by Histonet. The Histonet archives is 
a godsend. I am not sure whether Facebook or Instagram offers the same.

Anyway, I'm off to play golf.


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: 01 October 2024 15:26
To: Histonet@lists.utsouthwestern.edu ; Bob 
Richmond 
Subject: Re: [Histonet] Treponema tissue blocks

Dear Bob,
Good to see that you're still histonetting!  Until I saw your recent message I 
was contemplating abandoning the web site after reading and occasionally 
contributing for more than 30 years.
The current abundance of emails addressed to "histopeeps" is insulting, 
especially to professionally qualified technologists. Relia, whoever you are, 
give us all a break.
We need more correspondence of the kind we had in earlier decades, about the 
technology.
Hopefully you and I are not the only remaining elderly histonetters.
Others, young or old, please show yourselves.
Cheers,  John Kiernan
= = =


From: Bob Richmond via Histonet 
Sent: September 24, 2024 3:46 PM
To: Histonet@lists.utsouthwestern.edu 
Subject: [Histonet] Treponema tissue blocks

In reply to Ni Nie's request:

The best source of Treponema pallidum control tissue used to be necrotic
liver from necropsies of syphilitic stillbirths. These were only 25 years
in the past when I was a pathology resident in the 1960s. Possibly some of
this material is still around. As congenital syphilis returns, there might
be such material available today - one such case could supply the world for
years, after all.

Mycobacterium tuberculosis control tissue can be obtained from necropsies
on monkeys with tuberculosis, most commonly rhesus macaques in India.

Bob Richmond
Maryville TN
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Re: [Histonet] Cryotomy help

2024-07-12 Thread Tony Henwood via Histonet
I have never come across this problem but as a suggestion:


  1.
Methanol is often used as an anti-freeze, so I am not surprised that cryotomy 
is difficult
  2.
Rinse tissues well in a cell culture medium such as RPMI or Hanks to remove as 
much of the methanol as possible (several changes at least).
  3.
Pat-dry and infuse with Cryo-gel (50/50 with water, then 70%, finally undiluted 
Cryo-gel). Place on rotor for an hour or so each, until the tissue sinks in the 
Cryo-gel.

It appears that OCT and sucrose gradients interfere with the Mass Spectrometry 
due to the high sugar content (see Snijders, M. L., Zajec, M., Walter, L. A., 
de Louw, R. M., Oomen, M. H., Arshad, S., ... & Clahsen-van Groningen, M. C. 
(2019). Cryo-Gel embedding compound for renal biopsy biobanking. Scientific 
reports, 9(1), 15250.)

Lots of luck


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: Colleen Forster via Histonet 
Sent: 13 July 2024 03:19
To: Davoli, Katherine A 
Cc: histonet@lists.utsouthwestern.edu 
Subject: Re: [Histonet] Cryotomy help

Once the tissues are in any ethanol or methanol you will never be able to
get a frozen section. The only way I have been able to salvage anything
like this was to place the OCT block into 10%BNF on a rotator and let them
thaw/fix and process into a FFPE block.

If they cannot use FFPE because of the experiment I would say they need to
start over. This time, freeze the sample, NO METHANOL FIRST. Do the
fixation as  a post fix OR fix in an aqueous fixative such as 4%para or
10%BNF, sucrose protect and then freeze.

Believeme, I have tried to get this to work in so many ways and all experts
I consulted told me the exact same thing. The mehthaol will not allow the
sample to freeze hard enough to cut. They don't even want to use a slurry
with methanol to freeze it the OCT block. The tissue will  absorb enough of
that methanol to prevent good frozen sectioning.

Good Luck

Colleen Forster HT(ASCP)QIHC

On Fri, Jul 12, 2024 at 11:09 AM Davoli, Katherine A via Histonet <
histonet@lists.utsouthwestern.edu> wrote:

> The initial freeze isn't usually the problem.  It's what happens when it
> warms back up to Cryostat cutting temperatures of -20 or -25.
> You'd need to get the methanol out of the tissue completely because it
> causes the support to melt at the temperatures the Cryostat cuts at.
>
> And unfortunately I have not found a method that fully removes ethanol or
> methanol from a tissue sample that would then allow me to freeze and cut
> it.  If someone else replies with one I will genuinely be delighted because
> I have this problem a lot.
>
> Katherine Davoli, MDiv, HTL(ASCP)cm(they/them/theirs)
> Lab Manager, Tissue Culture & Histology Cores, U. Pitt Dept of
> Ophthalmology
> 7.373 UPMC Mercy Pavilion1622 Locust St.,Pittsburgh PA 15219
> (412) 624-8508   this number cannot receive texts
>
> -Original Message-
> From: Charles Riley via Histonet 
> Sent: Friday, July 12, 2024 12:04 PM
> To: histonet@lists.utsouthwestern.edu
> Subject: [Histonet] Cryotomy help
>
> I have a researcher who is trying to mass spectrometry on frozen tissue
> sections. They submitted the sample in methanol and even after sitting in a
> -80 freezer overnight the samples are too soft to section.
>
> Does anyone have any tips on how to harden the tissue for sectioning that
> won't damage the results for mass spect (I was thinking liquid nitrogen but
> want to be sure it won't damage the results)?
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--
Colleen Forster HT(ASCP)QIHC
BLS Histology and IHC Laboratory
Jackson Hall, Room 2-155
321 Church St. SE
Minneapolis, MN 55455
612-626-1930
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Re: [Histonet] cloudy cornea after Hartman's fix

2024-06-27 Thread Tony Henwood via Histonet
Hartman's (also known as Davidson's) fixative is sometimes used to reveal lymph 
nodes in resections where they appear opaque - white. It is not surprising that 
the tissues of the eye would react the same.
I assume that the alcohol causes the bleaching of the tissue (or is it the 
acetic acid?) - more research needed.


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: Davoli, Katherine A via Histonet 
Sent: 28 June 2024 04:12
To: histonet@lists.utsouthwestern.edu 
Subject: [Histonet] cloudy cornea after Hartman's fix

Anyone know why the cornea of my pig eye got white/cloudy on dropping it in to 
Hartman's fixative?  I'm used to working with mice where, if this happens I 
didn't notice.

Katherine Davoli, MDiv, HTL(ASCP)cm(they/them/theirs)
Lab Manager, Tissue Culture & Histology Cores, U. Pitt Dept of Ophthalmology
7.373 UPMC Mercy Pavilion1622 Locust St., Pittsburgh PA 15219
(412) 624-8508   this number cannot receive texts
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Re: [Histonet] Inquiry on Tissue Softening for Microtomy

2024-05-12 Thread 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

2024-05-01 Thread 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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> 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

2024-03-06 Thread 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

2024-02-08 Thread 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

2024-02-02 Thread 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&E tissue section

2023-11-10 Thread 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&E 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&E’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&E staining. This
>> might improve the H&E 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&E tissue section
>>
>>
>>
>> Hello,
>> I would like to know how to effectively restain a faded H&E 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&E tissue section

2023-11-09 Thread 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&E staining. This might improve 
the H&E 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&E tissue section

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

2023-09-13 Thread 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

2023-07-02 Thread 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

2023-07-01 Thread 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

2023-06-02 Thread 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:

> li {display:list-item;text-indent: -1em;}
> ul, ol{margin-left: 40px !important; padding-left: 0px
> !important;}
>
> When workloads spike, unexpected projects hit, employees are out
> for PTO, Sick or Parental Leave, we got your Lab Covered! Our
> Staffing Services for the lab can get the work done quickly. We
> have skilled laboratory talent at the ready and range of
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> and 5-10 years of experience
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> * Seeking Temporary or Permanent work
> * Location seeking: Houston, TX
> * Available: April
>
> *If you do not see a professional on this list that fi

Re: [Histonet] Sudanblack B on FFPET

2023-03-28 Thread 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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