[Histonet] Eosin in Alcohol

[Harrison, Sandra C.]

"polycyclic aromatic flourescent compounds that in high concentrations"
????  I wouldn't think the 3 mls of eosin dropped in the last 95%
alcohol could be considered "high concentration" but that's what keeps
Histonet entertaining; I learn something new every day.  

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Jennifer
Johnson
Sent: Tuesday, July 14, 2009 10:00 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Eosin in Alcohol


A couple of weeks ago I posted the message below on the histonet and all
of you responded that it shouldn't matter so I have finally gotten a
reply from the company we send our prostate biopsies off to and below is
their response.  So now you know the rest of the story!

 

We have used Eosin in the last 95% alcohol on the tissue processor for
several years. I usually add approximately 5 ml to the full jug. It is a
great tool to use for embedding. However, we received a letter from the
lab that we send our prostate biopsies to saying that it was undesirable
because it interfered with their immuno staining. They sent us some
cobalt blue to use in the place of eosin along with mixing instructions
and the whole batch of tissues came out such a dark blue. There is no
delineations in the color of the blue and I found it to be useless for
helping to embed. I would rather do without anything than use cobalt
blue. I guess the point of my rambling is, Eosin is a wonderful tool to
use unless you are doing immunos on prostate biopsies. 
 
Thanks,
 
Jennifer Johnson, HTL (ASCP) 


Their reply:  "The problem is that eosin belongs to a family of
polycyclic aromatic flourescent compounds that in high concentrations
binds to and saturates all tissue  components.  When immunoflourescence
is performed on such tissue- as in the prostate px+ test- the diffuse
background autoflourescence signal from prior treatment with these
compounds can interfere with, and even totally overwhelm, the signal of
the flourescent-labeled antibodies used to localize biomarkers in the
tissue."

 

 

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