Hi everyone,
I apologize if this has been asked before. I am running a study with rats and want to collect intestinal samples to look at histologically. Firstly, do the intestines need to be fixed in PFA before they are sectioned and stained? Secondly, many papers I see use H&E staining, but I am wondering if cresyl violet can be substituted instead?
Thank you.
You can do it, but it may not reveal the features you would hope to see.
See the figure here:
https://openi.nlm.nih.gov/detailedresult.php?img=PMC2746178_1476-4598-8-73-3&req=4
Hello
PFA is too "hard" fixative for the intestine so it is much better to use Bouin’s Fixative http://www.polysciences.com/skin/frontend/default/polysciences/pdf/866.pdf
The other thing is to use a matrix for the intestine - we use millipore disk, it is rather thin for cutting together with the specimen. If you use the long (1 cm) piece , cut it through wash with PBS for the removal of the content and then install onto the filter with the villus up. Put it into the cassete and then fix.
It is necessary to determine the direction of the section: the longitudinal section is preferable. If you use cresyl violet you may see mucin (but it usually is washed out )DNA and RNA, but it is better to use cresyl violet together with Luxol fast blue acc. Kluwer - Barrera kit
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