Re: [Histonet] IF with permanent mounting media?

[Morken, Timothy via Histonet]

Thanks John! This looks interesting to say the least.

Tim Morken
Supervisor, Electron Microscopy/Neuromuscular Special Studies
Department of Pathology
UC San Francisco Medical Center

From: John Kiernan 
Sent: Friday, May 28, 2021 3:20 PM
To: Histonet ; Morken, Timothy 

Subject: Re: IF with permanent mounting media?

Espada J, Juarranz A, Galaz S, Canete M, Villanueva A, Pacheco M, Stockert JC 
(2005) Non-aqueous permanent mounting for immunofluorescence microscopy. 
Histochem. Cell Biol. 123: 329-334. https://doi.org/10.1007/s00418-005-0769-2 ‍ 
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Espada J, Juarranz A, Galaz S, Canete M, Villanueva A, Pacheco M, Stockert JC 
(2005) Non-aqueous permanent mounting for immunofluorescence microscopy. 
Histochem. Cell Biol. 123: 329-334.  
https://doi.org/10.1007/s00418-005-0769-2
A PDF can be downloaded from the Google Scholar entry for this article.

John Kiernan
London, Canada
= = =

From: Morken, Timothy via Histonet 
mailto:histonet@lists.utsouthwestern.edu>>
Sent: May 28, 2021 11:31 AM
To: Histonet 
mailto:histonet@lists.utsouthwestern.edu>>
Subject: [Histonet] IF with permanent mounting media?

Has anyone tried using xylene/permanent mounting media for immunofluorescence 
stains? I had a question from a pathologist who wondered if we could do this. I 
have never heard of anyone doing it.

Tim Morken
Supervisor, Electron Microscopy/Neuromuscular Special Studies
Department of Pathology
UC San Francisco Medical Center

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Re: [Histonet] IF with permanent mounting media?

[John Kiernan via Histonet]

Espada J, Juarranz A, Galaz S, Canete M, Villanueva A, Pacheco M, Stockert JC 
(2005) Non-aqueous permanent mounting for immunofluorescence microscopy. 
Histochem. Cell Biol. 123: 329-334.  https://doi.org/10.1007/s00418-005-0769-2
A PDF can be downloaded from the Google Scholar entry for this article.

John Kiernan
London, Canada
= = =

From: Morken, Timothy via Histonet 
Sent: May 28, 2021 11:31 AM
To: Histonet 
Subject: [Histonet] IF with permanent mounting media?

Has anyone tried using xylene/permanent mounting media for immunofluorescence 
stains? I had a question from a pathologist who wondered if we could do this. I 
have never heard of anyone doing it.

Tim Morken
Supervisor, Electron Microscopy/Neuromuscular Special Studies
Department of Pathology
UC San Francisco Medical Center

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[Histonet] Two University of Wisconsin Positions

[Sandra Cheasty via Histonet]

Hello,
   We have two positions open in anatomic pathology at the School 
of Veterinary Medicine at the University of Wisconsin, Madison, the links are 
below...

https://jobs.hr.wisc.edu/en-us/job/509270/histology-technology-supervisor  
(application deadline June 10th)

https://jobs.hr.wisc.edu/en-us/job/509189/anatomic-pathology-laboratory-manager 
(application deadline June 3rd)

Sandy

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




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Re: [Histonet] IF with permanent mounting media?

[Ho-chun LAI via Histonet]

I personally did not tried FITC, but I'd used Cy dyes, Alexa Fluors, AF dyes as 
well as DAPI. I did not observe any harsh effect on fluorescence on those dyes. 
I'll guess FITC will also give good signal.

John Lai

Posdoctoral Fellow, School of Life Science, HKUST

-Original Message-
From: Morken, Timothy  
Sent: Friday, May 28, 2021 11:57 PM
To: Ho-chun LAI 
Cc: Histonet 
Subject: RE: IF with permanent mounting media?

Sounds interesting. How about with FITC label?

Tim Morken
Supervisor, Electron Microscopy/Neuromuscular Special Studies Department of 
Pathology UC San Francisco Medical Center

-Original Message-
From: Ho-chun LAI via Histonet 
Sent: Friday, May 28, 2021 8:54 AM
To: histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] IF with permanent mounting media?

Tim,



Yes it is possible. I added a distilled water rinse and air dry step after the 
final wash so the slide became dehydrated enough to be mounted by DPX.

JacksonImmuno does suggest Cy dyes for DPX mounts, but I found as long as you 
don't do the alcohol dehydration, alexa is far brighter.

The fluorescent signal will indeed be much more long lived than temporary mount 
like mowiol, some of my first mounts (GAD67/GFAP) retains signal even after an 
entire year. But you should expect they would fade somewhere around 3 months.



I realized I did not reply to the histonet the last time, I still need to get 
myself familiarized with this : (

John Lai

Posdoctoral Fellow, School of Life Science, HKUST



-Original Message-
From: Morken, Timothy via Histonet 
Sent: Friday, May 28, 2021 11:31 PM
To: Histonet 
Subject: [Histonet] IF with permanent mounting media?



Has anyone tried using xylene/permanent mounting media for immunofluorescence 
stains? I had a question from a pathologist who wondered if we could do this. I 
have never heard of anyone doing it.



Tim Morken

Supervisor, Electron Microscopy/Neuromuscular Special Studies Department of 
Pathology UC San Francisco Medical Center



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Re: [Histonet] IF with permanent mounting media?

[Morken, Timothy via Histonet]

Sounds interesting. How about with FITC label?

Tim Morken
Supervisor, Electron Microscopy/Neuromuscular Special Studies
Department of Pathology
UC San Francisco Medical Center

-Original Message-
From: Ho-chun LAI via Histonet  
Sent: Friday, May 28, 2021 8:54 AM
To: histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] IF with permanent mounting media?

Tim,



Yes it is possible. I added a distilled water rinse and air dry step after the 
final wash so the slide became dehydrated enough to be mounted by DPX.

JacksonImmuno does suggest Cy dyes for DPX mounts, but I found as long as you 
don't do the alcohol dehydration, alexa is far brighter.

The fluorescent signal will indeed be much more long lived than temporary mount 
like mowiol, some of my first mounts (GAD67/GFAP) retains signal even after an 
entire year. But you should expect they would fade somewhere around 3 months.



I realized I did not reply to the histonet the last time, I still need to get 
myself familiarized with this : (

John Lai

Posdoctoral Fellow, School of Life Science, HKUST



-Original Message-
From: Morken, Timothy via Histonet 
Sent: Friday, May 28, 2021 11:31 PM
To: Histonet 
Subject: [Histonet] IF with permanent mounting media?



Has anyone tried using xylene/permanent mounting media for immunofluorescence 
stains? I had a question from a pathologist who wondered if we could do this. I 
have never heard of anyone doing it.



Tim Morken

Supervisor, Electron Microscopy/Neuromuscular Special Studies Department of 
Pathology UC San Francisco Medical Center



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Re: [Histonet] IF with permanent mounting media?

[Ho-chun LAI via Histonet]

Tim,



Yes it is possible. I added a distilled water rinse and air dry step after the 
final wash so the slide became dehydrated enough to be mounted by DPX.

JacksonImmuno does suggest Cy dyes for DPX mounts, but I found as long as you 
don't do the alcohol dehydration, alexa is far brighter.

The fluorescent signal will indeed be much more long lived than temporary mount 
like mowiol, some of my first mounts (GAD67/GFAP) retains signal even after an 
entire year. But you should expect they would fade somewhere around 3 months.



I realized I did not reply to the histonet the last time, I still need to get 
myself familiarized with this : (

John Lai

Posdoctoral Fellow, School of Life Science, HKUST



-Original Message-
From: Morken, Timothy via Histonet 
Sent: Friday, May 28, 2021 11:31 PM
To: Histonet 
Subject: [Histonet] IF with permanent mounting media?



Has anyone tried using xylene/permanent mounting media for immunofluorescence 
stains? I had a question from a pathologist who wondered if we could do this. I 
have never heard of anyone doing it.



Tim Morken

Supervisor, Electron Microscopy/Neuromuscular Special Studies Department of 
Pathology UC San Francisco Medical Center



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Re: [Histonet] IF with permanent mounting media?

[Paula Keene Pierce via Histonet]

Hi,
you can use "Crystal Mount" first, let it dry, and then coverslip with 
xylene/resin.
I do this routinely with AEC IHC and oil red O and other fat stains.
Crystal Mount only on retinal flat mounts to be shipped out to others for 
future staining, as it can be removed with water,

FYI: Crystal mount is no longer made under that name, but do a search on 
Electron Microscopy Sciences website and their version will come up in the 
results.
Paula Keene Pierce, BS, HTL(ASCP)HTPresidentExcalibur Pathology, Inc.5830 N 
Blue Lake DriveNorman, OK 73069PH 405-759-3953http://www.excaliburpathology.com

A sharp knife is nothing without a sharp eye. - Klingon Proverb 

On Friday, May 28, 2021, 10:43:39 AM CDT, Morken, Timothy via Histonet 
 wrote:  
 
 Has anyone tried using xylene/permanent mounting media for immunofluorescence 
stains? I had a question from a pathologist who wondered if we could do this. I 
have never heard of anyone doing it.

Tim Morken
Supervisor, Electron Microscopy/Neuromuscular Special Studies
Department of Pathology
UC San Francisco Medical Center

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[Histonet] IF with permanent mounting media?

[Morken, Timothy via Histonet]

Has anyone tried using xylene/permanent mounting media for immunofluorescence 
stains? I had a question from a pathologist who wondered if we could do this. I 
have never heard of anyone doing it.

Tim Morken
Supervisor, Electron Microscopy/Neuromuscular Special Studies
Department of Pathology
UC San Francisco Medical Center

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