This is from the "Theory and Practice of HIstotechnology", 2nd edition, 190.
Dezna C. Sheehan and Barbara B. Hrapchak. The reference for their procedure
is the article you mentioned.
FIXATIVE:
Mercuric formalin preferred, but 10% NBF can be used
SOLUTIONS:
Mayer Hemaxylin
Phloxine stain
- 1 g Phloxine
- 200 m of 70% Ethanol
- 1 g Calcium chloride
Tartrazine solution
- 2.5 g Tartrazine
- 100 mL cellosolve (ethylene glycol monoethyl ether)
PROCEDURE:
1. Deparaffinize and hydrate slides to d. water
2. Stain for 5-10 minutes in Mayer hematoxylin
3. Blue sections in running tap water for 15 minutes
4. Stain with phloxine solution for 30 minutes
5. Rinse briefly in distilled water, and drain on filter paper
6. Differentiate with tartrazine solution until the inclusion bodies stand
out bright red and the background is yellow.
7. Dehydrate; clear in xylene; coverslip using a synthetic mounting media
RESULTS:
Inclusion bodies = red
Nuclei = blue
Background - yellow
NOTES FROM PEGGY:
1. Seems like you should be able to use Gill regressive or Harris
regressive, too. At the time we did this stain 30 years ago, our lab was
using Mayer hematoxylin.
2. It's very similar to doing a Brown and Hopps, or a Brown and Brenn. Not
enough time in the tartrazine differentiator, and the background will be
reddish, making is hard to find the inclusion bodies. Too much time in the
tartrazine differentiator, and the background is yellow but so are the
inclusion bodies. I remember doing this for something decades ago, and
remember how easy it was to over and under differentiate.
3. I seem to remember reusing the staining solutions multiple times.
4. Yes, we used cellosolve to make the tartrazine. It wasn't the only
staining solution that used it, so we had it around. I'm wondering if you
could use something like acetone instead, but do a much faster
differentiation.
Peggy A. Wenk, HTL(ASCP)SLS
-----Original Message-----
From: Wolfe, Christina
Sent: Monday, June 16, 2014 4:35 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Lendrum's stain for Paneth Cells
Hi,
Does anyone have a PMID for Lendrum's original paper " LENDRUM, A.C. 1947.
The Phloxine-tartrazine method as a general histological stain and for the
demonstration of inclusion bodies. Journal of Pathology and Bacteriology."?
I have searched PubMed and I am not getting any results. We are having
trouble getting this stain to work in our lab. The Paneth cells are not
staining and there is no yellow stain at all with the tartrazine. We are
trying to stain rat ileum. Anyone with experience using this stain - please
help. :)
Thanks!
Kristie
Christina Wolfe, BS, MLT (ASCP), HT, QIHC
Drug Safety Evaluation/Bristol-Myers Squibb
Pathology Dept.
812-307-2093
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