Re: [Histonet] Lendrum's stain for Paneth Cells

[Lee & Peggy Wenk]

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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Re: [Histonet] Lendrum's stain for Paneth Cells

[Grantham, Andrea L - (algranth)]

I just did this stain again and it was beautiful.  I used Lendrum's protocol 
from Stains File web site.

http://stainsfile.info/StainsFile/stain/micro/phloxtartlend.htm


When I got to step 5 I proceeded one slide at a time and checked under the 
microscope to get the differentiation to where I wanted it. The time varies for 
each slide.





Andrea Grantham, HT (ASCP)
Senior Research Specialist
University of Arizona
Cellular and Molecular Medicine
Histology Service Laboratory
P.O.Box 245044
Tucson, AZ 85724

algra...@email.arizona.edu
Tel: 520.626.4415 Fax: 520.626.2097





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