[Histonet] IF with permanent mounting media?

[Ho-chun LAI]

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 <Timothy.Morken at ucsf.edu> 
Sent: Friday, May 28, 2021 11:57 PM
To: Ho-chun LAI <hclaiac at connect.ust.hk>
Cc: Histonet <histonet at lists.utsouthwestern.edu>
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 <histonet at lists.utsouthwestern.edu>
Sent: Friday, May 28, 2021 8:54 AM
To: histonet at 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 <histonet at lists.utsouthwestern.edu>
Sent: Friday, May 28, 2021 11:31 PM
To: Histonet <histonet at 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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