Re: [Histonet] Agarose embedded tissue arrays embedded in paraffin block

[Bernice Frederick via Histonet]

Exactly, I process the agar,not the tissue in it Cells,organoids, spheroids.
Bernice

-Original Message-
From: Colleen Forster via Histonet  
Sent: Friday, May 17, 2024 3:07 PM
To: Jay Lundgren 
Cc: histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] Agarose embedded tissue arrays embedded in paraffin 
block

Chistopher,

Were these small pieces of tissue? If they are in an agar (such as
histogel) they would need to have been processed overnight to ensure the agar 
is completely dehydrated during processing. The sample, no matter how small, is 
protected and processes perfectly. IF you ran a short run with the agar it will 
not have [processed properly. I was never able to salvage those samples. It 
took me a couple times to figure out the solution.

Just a thought.

Colleen Forster HT(ASCP)QIHC

On Fri, May 17, 2024 at 2:32 PM Jay Lundgren via Histonet < 
histonet@lists.utsouthwestern.edu> wrote:

> Where did the agarose come from?
>
> On Fri, May 17, 2024 at 12:09 PM Otto, Christopher M via Histonet < 
> histonet@lists.utsouthwestern.edu> wrote:
>
> >
> >
> > Hello everyone!
> >
> >  I'm having trouble sectioning tissue array blocks where the array 
> > is in agarose embedded into a paraffin block.  I've chilled the 
> > blocks and I'm sectioning on a rotary microtome, at 5 microns, with 
> > a high profile Accuedge blade. The paraffin surrounding the agarose 
> > sections normally,
> but
> > the agarose portion of the block causes the blade to "skip" across 
> > it slightly and even chip out as if my blade isn't snug in the blade 
> > holder (it is).  If I do get a tiny portion of agarose on my section 
> > to float
> out
> > on the waterbath it flies away (like adding ETOH to a waterbath with
> > sections already on it.)   Everyone I have asked about this says the
> > agarose should section normally with the paraffin like any other 
> > FFPE blocks. Any ideas on why this agarose is behaving this way for 
> > me?  Thank you in advance!
> >
> >
> >
> > ___
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> EB3125Zq$
>


--
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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[Histonet] FW: Golgi-cox staining

[Bernice Frederick via Histonet]

From: Rizaldy P Scott 
Sent: Wednesday, April 10, 2024 4:33 PM
To: Bernice Frederick 
Subject: Re: [Histonet] Golgi-cox staining

Hi Bernice,

What they should have done ideally is to store the uncut agarose blocks 
submerged in whatever buffer they used to rinse the brains out prior to agarose 
embedding. They can probably salvage the dried-out brains by rehydrating in 
their buffer or 30% sucrose solution overnight (or store at 4 degrees as 
needed) and re-embed in agarose only when they are ready to cut. I can't 
guarantee however if artefacts due to drying might result but it's worth the 
shot.


Best regards,
Riz

Rizaldy P. Scott, M.S., Ph.D.
Research Associate Professor of Pathology
Scientific Director
Mouse Histology & Phenotyping Laboratory
Robert H. Lurie Comprehensive Cancer Center
Northwestern University Feinberg School of Medicine
Olson Pavilion, Room 8-335, 710 N. Fairbanks Ct., Chicago, IL 60611
✉️  rizaldy.sc...@northwestern.edu
 +1-(312)-503-2695
https://www.feinberg.northwestern.edu/sites/mhpl/
www.cancer.northwestern.edu



From: Bernice Frederick 
mailto:b-freder...@northwestern.edu>>
Sent: Wednesday, April 10, 2024 13:40
To: Rizaldy P Scott 
mailto:rizaldy.sc...@northwestern.edu>>
Subject: FW: [Histonet] Golgi-cox staining

Can you help this person?
Bernice

-Original Message-
From: Mariela Chertoff via Histonet 
mailto:histonet@lists.utsouthwestern.edu>>
Sent: Wednesday, April 10, 2024 9:28 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Golgi-cox staining

Hi all

We made the Golgi cox staining and  due to a problem with the vibratome, we 
left the tissue several days embedded in  agarose and they get dryed, It is 
possoble to recover the brains? it is better to repeat the agarose embebbing o 
it is better to put the brains in crioprerervate solution to rehidrated and 
after that put them in agarosa again? We are following the Zaquot protocol 
https://urldefense.com/v3/__https://www.frontiersin.org/articles/10.3389/fnana.2016.00038/full__;!!Dq0X2DkFhyF93HkjWTBQKhk!QkwV4_RJl5cyhElJgSA-8lRMmOSmkwVoHzRrMg8-ZXGX0yUzNCxM_pm09t45KaZWo04kpjD-tBduP59_0DHjxeEvlRMxvl848hoVaCzp$

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 5285-8680/1/2
email:marielachert...@gmail.com
marielachert...@qb.fcen.uba.ar
mailto:mariela.chert...@uab.cat>>
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Re: [Histonet] preordering giemsa stains

[Bernice Frederick via Histonet]

We used to Alcian Yellow...
Bernice

-Original Message-
From: Gudrun Lang via Histonet  
Sent: Thursday, February 29, 2024 8:48 AM
To: 'Paula' 
Cc: histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] preordering giemsa stains

In my place it is ordered on demand, but the pathologists usually perfer IHC 
for Hp.

Gudrun

-Ursprüngliche Nachricht-
Von: Paula via Histonet [mailto:histonet@lists.utsouthwestern.edu]
Gesendet: Dienstag, 27. Februar 2024 20:46
An: histonet@lists.utsouthwestern.edu
Betreff: [Histonet] preordering giemsa stains

Hello,

Is it a common practice to preorder stains that involve the stomach searching 
for H pylori?  Or, is it more common to order as needed?

Thank you for anyone's feedback.

Paula Lucas

Lab Manager

 

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Re: [Histonet] Von Kossa staining

[Bernice Frederick via Histonet]

I use a 60 watt bulb and it works fine.
Bernice

-Original Message-
From: John Kiernan via Histonet  
Sent: Wednesday, July 26, 2023 11:51 PM
To: Charles Riley ; histonet@lists.utsouthwestern.edu; dsiena 

Subject: Re: [Histonet] Von Kossa staining


Charles,

A handheld light of any kind isn't really suitable because you would have to 
hold it over the slides for 15 to 60 minutes, according to which variant of the 
von Kossa method you plan to use (see Lillie & Fullmer 1976 Histopathologic 
Technic ... 4th ed. pp 539-541).

An anglepoise lamp with an old-fashioned 100W bulb is OK, and so is a sunny 
windowsill. Silver salts absorb at the blue end of the spectrum, so a 
fluorescent light should be more efficient than an incandescent bulb. If no 
bright light source is available, it's possible to chemically reduce the silver 
phosphate and/or carbonate to black colloidal silver, with a traditional 
photographic developer. The method of Rungby et al.1993 may be better than 
other post-reduction methods 
(https://urldefense.com/v3/__https://scholar.google.ca/scholar?hl=en_sdt=0*2C5=rungby*1993*calcium*deposits=rungby*1993__;JSsrKys!!Dq0X2DkFhyF93HkjWTBQKhk!SGP1gx0IglGW5nnid6Tbv6aRuFAWHsIiz6kn3jDzSw7njXRiBLURbNbbv4YCJKl-Jbcpm60EOukGFQIdqh2rLhqFIgrWGAWfChgoE5Zi$
 ). I never tried it, but Rungby's paper has collected 104 citations, which is 
very good for a paper in our field.

The von Kossa technique is simply explained in my Histological and 
Histochemical Methods textbook, 5th edn (2015). The book costs less than 1ml of 
any antibody.

Enough said!   John
John A. Kiernan
Emeritus, Anatomy & Cell Biology,
University of Western Ontario
London, Canada
https://urldefense.com/v3/__https://www.schulich.uwo.ca/anatomy/people/bios/emeriti/kiernan_john.html__;!!Dq0X2DkFhyF93HkjWTBQKhk!SGP1gx0IglGW5nnid6Tbv6aRuFAWHsIiz6kn3jDzSw7njXRiBLURbNbbv4YCJKl-Jbcpm60EOukGFQIdqh2rLhqFIgrWGAWfCiLlysL2$
 
Also  Secretary, Biological Stain Commission, Inc.
https://urldefense.com/v3/__https://biologicalstaincommission.org__;!!Dq0X2DkFhyF93HkjWTBQKhk!SGP1gx0IglGW5nnid6Tbv6aRuFAWHsIiz6kn3jDzSw7njXRiBLURbNbbv4YCJKl-Jbcpm60EOukGFQIdqh2rLhqFIgrWGAWfCtBsy2U8$
 
= = =








-Original Message-
From: Charles Riley via Histonet 
Sent: Wednesday, July 26, 2023 1:23 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Von Kossa staining


*** Externally sourced email message ***


Can anyone out there who performs Von Kossa staining provide me with any 
guidelines or suggestions for the light source to use for the Silver nitrate 
activation?

Is a standard handheld black light strong enough or does it need to be a UV 
sanitizing strength light if using UV versus incandescent bulb exposure?
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[Histonet] water under sectios

[Bernice Frederick via Histonet]

Good morning (well, in Chicago anyway)
We've recently been having issues with water under the sections when picked up 
from the waterbath. It seems to even affect the sections during antigen 
retrieval after removing excess water, drying and baking I was given a block 
from an outside source that I know is a different paraffin and there was no 
water to be seen. Same person,same techniques. Can paraffin play a part? Or is 
the processing throwing this off. We run various programs and it seems to 
happen with almost everything. TMA"s in particular, as cores get lost during 
retrieval.
Bernice

Bernice Frederick
Pathology Core Facility
Robert H. Lurie Cancer Center
710 North Fairbanks Court
Olson 8-421
312-503-3723
b-freder...@northwestern.edu

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[Histonet] RAt teeth

[Bernice Frederick via Histonet]

Hi all,
I have bamboo rat teeth that have (ha-ha) been decalled. Obviously they are not 
soft. Can I use Nair on them or is there something better?  I will need an H 
and an iron stain. They are not tiny samples..
Bernice

Bernice Frederick
Pathology Core Facility
Robert H. Lurie Cancer Center
710 North Fairbanks Court
Olson 8-421
312-503-3723
b-freder...@northwestern.edu

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[Histonet] P16

[Bernice Frederick via Histonet]

Who is the best and current supplier for p16? We were using biocare,but 
evidently it is no longer available from them. And no Santa Cruz per my IHC 
tech.
Thanks,
Bernice

Bernice Frederick
Pathology Core Facility
Robert H. Lurie Cancer Center
710 North Fairbanks Court
Olson 8-421
312-503-3723
b-freder...@northwestern.edu

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Re: [Histonet] sectioning issues?

[Bernice Frederick via Histonet]

You may have to call a tech at thermos to ask.. It still sounds like it 
it's the microtome as we know they cut. Join Histonet or the NSH facebook page 
and you can ask there as well. I may just be missing something obvious. 
histonet@lists.utsouthwestern.edu for 
histonet. It is a forum.
Bernice
Bernice Frederick HTL (ASCP)
Senior Research Tech
Pathology Core Facility
Robert. H. Lurie Cancer Center
Northwestern University
710 N Fairbanks Court
Olson 8-421
Chicago,IL 60611
312-503-3723
b-freder...@northwestern.edu

From: Elizabeth Jane Lux
Sent: Friday, April 21, 2017 10:04 AM
To: Bernice Frederick
Subject: Re: sectioning issues?


Thanks so much for your help!

Frustratingly having issues when I move into sectioning. Ribbons look square at 
trim mode (20u) but when I try to get my sections, I'm getting inconsistent 
slicing, almost like it's compressing it at times. Or I guess not advancing the 
specimen correctly? Perhaps the arm needs maintenance?



Oh, and I was completely wrong on the model, it's a Thermo Finesse ME+



Liz

From: Elizabeth Jane Lux
Sent: Friday, April 21, 2017 8:58 AM
To: Bernice Frederick
Subject: Re: sectioning issues?


Awesome! I'll bring the block(s) over now, thanks!



Elizabeth Lux
Research Tech 3
Feinberg Cardiovascular Research Institute
Northwestern University
Lurie Building, 10-220
312.503.6368 lab phone
312.503.0219 fax

From: Bernice Frederick
Sent: Friday, April 21, 2017 8:55 AM
To: Elizabeth Jane Lux
Subject: RE: sectioning issues?

You can always pop by and let me see. Now would be fine. I am gone all next 
week.
Bernice

Bernice Frederick HTL (ASCP)
Senior Research Tech
Pathology Core Facility
Robert. H. Lurie Cancer Center
Northwestern University
710 N Fairbanks Court
Olson 8-421
Chicago,IL 60611
312-503-3723
b-freder...@northwestern.edu

From: Elizabeth Jane Lux
Sent: Friday, April 21, 2017 8:27 AM
To: Bernice Frederick
Subject: Re: sectioning issues?


Tissue is kidney and heart. Thickness was going off of past work done in the 
lab, utilizing the guidelines in the AFIP Lab methods in Histo red book.

The machine had some maintenance done last year, though the tech seemed really 
green and unsure of himself.

The block seems secure to the cassette.



Thanks for your thoughts!



Elizabeth Lux
Research Tech 3
Feinberg Cardiovascular Research Institute
Northwestern University
Lurie Building, 10-220
312.503.6368 lab phone
312.503.0219 fax

From: Bernice Frederick
Sent: Friday, April 21, 2017 7:43 AM
To: Elizabeth Jane Lux
Subject: RE: sectioning issues?

Hi Liz,
What kind of tissue are you sectioning? Bear in mind,the microtome you are 
using may have never had any maintenance on it. And why are you cutting at 6? 
Unless you are doing neuro or looking for amyloid, 4 um is the best thickness. 
The block itself may be coming away from the cassette. This can cause said 
issues. Wiggle the paraffin and see if it is not coming away from the cassette.
Bernice

Bernice Frederick HTL (ASCP)
Senior Research Tech
Pathology Core Facility
Robert. H. Lurie Cancer Center
Northwestern University
710 N Fairbanks Court
Olson 8-421
Chicago,IL 60611
312-503-3723
b-freder...@northwestern.edu

From: Elizabeth Jane Lux
Sent: Thursday, April 20, 2017 4:09 PM
To: Bernice Frederick
Subject: sectioning issues?


Hey Bernice,

Wonder if I could ask your opinion on some microtome troubleshooting. I'm 
having a few tricky paraffin blocks. Facing it gets me nice smooth ribbons, but 
when I move to the sections (6micron) I get inconsistent results. It will give 
me a full section, then a half, then full, then half, and so on . .

Moved to a test block to work out the kinks, and adjusted the angle slightly, 
tightened up the blade holder, and things seemed to be ribboning better. But 
then still having issues with the sample block of interest.

Anyway, thought it was worth asking if you had any advice.

Thanks!

Liz

Elizabeth Lux
Research Tech 3
Feinberg Cardiovascular Research Institute
Northwestern University
Lurie Building, 10-220
312.503.6368 lab phone
312.503.0219 fax
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[Histonet] TUNEL

[Bernice Frederick via Histonet]

Hello all.
Would there be any reason that slides stained with TUNEL (ISH) would not pick 
up the Mayers counterstain? We use it for IHC and it is fine. Mayers for 7 
minutes and then ammonia water for 1 minute. We do that rather than rinse for 
15 minutes and there are usually no issues. Could a reagent in the stain be 
causing it? DAPI is used...
Bernice

Bernice Frederick HTL (ASCP)
Senior Research Tech
Pathology Core Facility
Robert. H. Lurie Cancer Center
Northwestern University
710 N Fairbanks Court
Olson 8-421
Chicago,IL 60611
312-503-3723
b-freder...@northwestern.edu

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[Histonet] Fibroblasts

[Bernice Frederick via Histonet]

Hello all,
Don't have a ton of time to search so am asking. Any good markers specific to 
fibroblasts? And am drawing a blank for regular special stains for some reason. 
Researcher says Vimentin is not specific enough.
Thanks,
Bernice

Bernice Frederick HTL (ASCP)
Senior Research Tech
Pathology Core Facility
Robert. H. Lurie Cancer Center
Northwestern University
710 N Fairbanks Court
Olson 8-421
Chicago,IL 60611
312-503-3723
b-freder...@northwestern.edu

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Re: [Histonet] floor vibration

[Bernice Frederick via Histonet]

We have issues as HVAC is above us! Sometimes at the switch over from heat to 
air and vice versa the facilities personnel don't check the drains and we get a 
flood. Came in one morning to hear dripping and we were going to a self -eval 
that day.! Most of the water was in my garbage can (Thankfully) Just missed my 
PC and the TMA arrayer. Got part of the embedding unit (it was a big leak). 
Facilities had the wet/dry vac up in the ceiling to get rid of the water. Had 
to have a good chunk of ceiling replaced and repaired after that one. There was 
even water in the light fixtures!
Bernice

Bernice Frederick HTL (ASCP)
Senior Research Tech
Pathology Core Facility
Robert. H. Lurie Cancer Center
Northwestern University
710 N Fairbanks Court
Olson 8-421
Chicago,IL 60611
312-503-3723
b-freder...@northwestern.edu

-Original Message-
From: Jeffrey Robinson via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Sent: Monday, May 16, 2016 5:08 PM
To: histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] floor vibration

An additional word of caution about placing the EM Lab under ANY kind of 
potential water leak.  Tim Morken and I can attest to that!  They installed a 
new CT scanner in Radiology directly above the EM Lab.  They broke some sort of 
water pipe during the installation and it leaked so much that part of the 
ceiling collapsed onto the scope!  They didn't bother checking to see if there 
was any damage on the next floor down- they just left.  Tim had a huge surprise 
waiting for him the next morning.  We always had to cover the scope with a 
sheet of plastic at night after that in case they installed something else.  
Jeff Robinson, Senior Histotechnologist, EM Tech (emeritus), Sierra Pathology 
Lab, Clovis, CA.

-Original Message-
From: Keyser Gerald T via Histonet [mailto:histonet@lists.utsouthwestern.edu]
Sent: Monday, May 16, 2016 1:25 PM
To: 'Nancy Schmitt'; histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] floor vibration

I can think of two things.

First, relentlessly make fun of your building planner for putting a histology 
lab underneath a laundry. This is a mistake worthy of pointing and laughing.

Second, there are isolation tables and platforms. That's probably the first 
thing I would try.

http://www.taab.co.uk/pdf-details/305_taab_products_1402580607.pdf


Gerry

-Original Message-
From: Nancy Schmitt via Histonet [mailto:histonet@lists.utsouthwestern.edu]
Sent: Monday, May 16, 2016 10:03 AM
To: 'histonet@lists.utsouthwestern.edu'
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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[Histonet] Mycoplasma

[Bernice Frederick via Histonet]

Hello all,
Will PAS stain mycoplasma?
Bernice

Bernice Frederick HTL (ASCP)
Senior Research Tech
Pathology Core Facility
Robert. H. Lurie Cancer Center
Northwestern University
710 N Fairbanks Court
Olson 8-421
Chicago,IL 60611
312-503-3723
b-freder...@northwestern.edu

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[Histonet] CAP

[Bernice Frederick via Histonet]

ANP21382. What kind of policy or SOP do you all have for this question? T asks 
how reagents are given an expiration. This includes but is not restricted to 
reagents where manufacturer does not specify a date. We date made up reagents 
as a 6 month expiration unless we know it doesn't last that long. Came up 
during interim self inspection.
Thanks,
Bernice

Bernice Frederick HTL (ASCP)
Senior Research Tech
Pathology Core Facility
Robert. H. Lurie Cancer Center
Northwestern University
710 N Fairbanks Court
Olson 8-421
Chicago,IL 60611
312-503-3723
b-freder...@northwestern.edu

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[Histonet] CD34

[Bernice Frederick via Histonet]

Been looking around a bit and seem to have it a roadblock. Does anyone know of 
a vendor for CD34 that will cross react with rabbit? On frozen sections, mind 
you.
Thanks,
Bernice

Bernice Frederick HTL (ASCP)
Senior Research Tech
Pathology Core Facility
Robert. H. Lurie Cancer Center
Northwestern University
710 N Fairbanks Court
Olson 8-421
Chicago,IL 60611
312-503-3723
b-freder...@northwestern.edu

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[Histonet] Oil Red O

[Bernice Frederick via Histonet]

Hello all,
Will Oil Red O stain phospholipids? We will be helping another lab with some 
work and the project out line states that Phopslipids are externalized on the 
cell surface during apoptosis. Can always try it I suppose.
Bernice

Bernice Frederick HTL (ASCP)
Senior Research Tech
Pathology Core Facility
Robert. H. Lurie Cancer Center
Northwestern University
710 N Fairbanks Court
Olson 8-421
Chicago,IL 60611
312-503-3723
b-freder...@northwestern.edu

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[Histonet] Masson's on Frozens.

[Bernice Frederick via Histonet]

Question:
When we stain and HE on frozen we fix in 95% prior to staining. As I never in 
30 years have done a Masson's Trichrome on a  frozen section I ask : do I need 
to fix and then mordant as usual or go straight to the  Bouin's and continue as 
normal? Do I need to mordant at all? Tissue is rabbit liver.
BErnice

Bernice Frederick HTL (ASCP)
Senior Research Tech
Pathology Core Facility
Robert. H. Lurie Cancer Center
Northwestern University
710 N Fairbanks Court
Olson 8-421
Chicago,IL 60611
312-503-3723
b-freder...@northwestern.edumailto:b-freder...@northwestern.edu

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