[Histonet] Some questions about dehydration and clearing
Allan Wang
allanvv at gmail.com
Sun Dec 4 01:23:58 CST 2016
Hello,
I've been doing some histology work but without formal training. Just some
questions about protocol:
1) Is it okay to dehydrate directly into 100% alcohol or does that cause
tissue shrinkage? Is starting at 95% good enough? I'm trying to reduce the
number of stations needed.
2) I've once been told that after dehydration, I should wait for all the
alcohol to dry off before clearing with xylene. Does that make sense, since
I couldn't find this step in any protocols I've seen. Or would this mess up
morphology?
Sometimes I see some inconsistent appearance of cells after IHC staining,
not sure if it's due to issues with this dehydration step/clearing or if
it's the change in antibody dilution that's causing it.
I have a comparison image here, where I think something went wrong with
Slide 2. Both IHC was performed with an autostainer.
http://i.imgur.com/y0q4hb7.jpg
Thanks,
Allan Wang
More information about the Histonet
mailing list