Hello Jennifer, yes over-fixation will cause adhesion problems. This applies to
formalin fixed frozens as well as PFA fixed frozens.
I have had good luck using chromated gelatin to coat slides. I have
successfully cut and stained lung specimens left in NBF for several months.
Commercially,
No...never add sucrose to "PFA" when fixing tissues for freezingor any
other time.
Fix, then sucrose- immerse until tissue sinks to bottom of receptacle if
you can't snap-freeze effectively
Carl Hobbs FIBMS
Histology and Imaging Manager
Wolfson CARD
Guys Campus, London
Good charged slides or subbed slides will work.
In regards to the fixation, I personally find the morphology with 4% PFA O/N
followed by 30% sucrose better than doing them simultaneously.
Michelle
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Histonet mailing list
Hi Jenny.
Can you use Pwax embedding as an alternative?
H will always be better on Pwax sections.
IHC will also be in many instances.
What abs are you probing with?
Single/double-labelling?
Yesyou are fixing for too long ( for frozen sectioningfor Pwax you can
fix your specimens for
Thank you Charles Scouton for responding. The slides are positive charged
slides from IMEB.
Thank you Elizabeth Chilpala for responding. The treatment after sectioning is
one I’ve forwarded to the service that is processing for us. I only do the
PFA/sucrose/embedding part. My understanding
]
Sent: Friday, July 29, 2016 1:48 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] PFA Fixed tissue not sticking to slides?
Dear HistoFolks,
Has anyone had experience with cryo-sectioned tissue that is PFA fixed not
adhering to slides?
The tissue is rabbit cornea culture, 4% PFA fixed O
Dear HistoFolks,
Has anyone had experience with cryo-sectioned tissue that is PFA fixed not
adhering to slides?
The tissue is rabbit cornea culture, 4% PFA fixed O/N, then run through 20%
sucrose gradient, then frozen in OCT.
We think that the PFA fix is much longer than needed for a tissue
