AW: [Histonet] bluing hematoxylin and alkaline water????
gu.lang <@t> gmx.at
Fri Nov 21 11:01:40 CST 2008
Is this a Haematoxylin-Eosin-stain? If so, I think you wash out the Eosin
during the long water-wash. That makes the difference.
Haemalaun binds to negative charged tissueparts best in acid solution
(pH3-4). With this pH the colour is redish. Blueing means to elevate the pH
about 7-8. Now the bound dye-molecule is remodeld. The colour turns blue and
is more stable.
Von: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] Im Auftrag von Eva
Gesendet: Freitag, 21. November 2008 17:11
An: histonet <@t> lists.utsouthwestern.edu
Betreff: [Histonet] bluing hematoxylin and alkaline water????
Good morning out in histoland,
Thank god it is Friday. Have just a small question that I am sure you
all have an answer for.
It is about bluing slides after hematoxylin. I read somewhere that
alkaline water can blue hematoxylin and I am wondering if this is part
of our problem. We use an automated stainer that we run both during the
day and during the night. Normally we blue the slides in 1% ammonium
after the runs for 1min. The run that was done during the day ended up
less blue (more purple). The ones that run during the night continue
being washed in water until we come in in the morning. They ended up
more blue. I tested the pH of the water we are using. It is 7.96. Would
this be enough to blue the slides if they are washed in it every hour
from 9pm to 8am? Any other reason this might be happening? Should we not
blue the slides that are run during the night? Why do we blue
Thank you for all your answers
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