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United States Government Work, uncopyrighted, public-domain,
DRAFT COPY ONLY. This document does not necessarily represent the views
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PRINCIPLE OF THE TEST.
To stain paraffinized tissue sections for Giemsa positivity, required
in the diagnosis of some diseases encountered in anatomic pathology,
especially Helicobacter in gastric biopsies.
SPECIMEN REQUIRED.
All human tissue excised at surgery, outpatient clinics, and postmortems,
fresh or in fixative, along with a filled-out Tissue Examination Form
(U. S. Standard Form 515, USSF515).
REAGENTS, INSTRUMENTATION.
Jenner solution (stock, working).
Jenner stain powder.
Methyl Alcohol.
Giemsa solution (stock, working).
Giemsa powder.
Glycerine.
1% Glacial acetic acid solution.
Glacial acetic acid.
Iodine.
Sodium thiosulfate.
95% alcohol.
Absolute alcohol.
Xylene substitute.
Permount or Histoclad.
STEP-BY-STEP DESCRIPTION.
FlXATION:
Zenker's or other well-fixed tissue.
TECHNIQUE:
Cut paraffin sections at 6 microns.
SOLUTIONS:
JENNER SOLUTION (STOCK).
Jenner stain............................... 1 gm.
Alcohol, methyl............................ 400 ml.
JENNER SOLUTION (WORKING)
Jenner solution (stock)................... 25 ml
Distilled water........................... 25 ml
GIEMSA SOLUTION (STOCK)
Giemsa powder............................. 1 gm
Glycerin.................................. 66 ml
Alcohol, methyl........................... 66 ml
Mix glycerin and Giemsa powder. Place in 60 o C oven for 2 hours.
Cool and add methyl alcohol.
GIEMSA SOLUTION (WORKING)
Giemsa solution (stock)................. 50 drops
Distilled water.......................... 50.0 ml
1% GLACIAL ACETIC WATER SOLUTION
Glacial acetic acid...................... 1.O ml
Distilled water......................... 100.0 ml
STAINING PROCEDURE.
1. Deparaffinize and hydrate to distilled water.
2. Remove mercuric chloride crystals with iodine and clear with
sodium thiosulfate.
3. Wash in running water for 10 minutes.
4. Rinse in distilled water, two changes.
5. Methyl alcohol, two changes for 3 minutes each.
6. Working Jenner solution for 6 minutes.
7. Working Giemsa solution for 45 minutes.
8. Handle each s1ide individually in this and subsequent steps.
Differentiate in glacial acetic water solution, then check
microscopically for well differentiated nuclei.
9. Rinse in distilled water.
10. Dehydrate quickly in 95% alcohol, absolute alcohol, and clear with
xylene substitute, two changes each.
11. Mount with Permount or Histoclad
CONTROL: Helicobacter gastritis skin
RESULTS.
Nuclei .......... blue.
Cytoplasm ....... pink to rose.
Bacteria ........ blue.
REFERENCES.
1. Swisher BL.
Microorganisms. Chapter 16, pp. 325-344.
In: Bancroft JD, Gamble M.
Theory and Practice of Histological Techniques. Fifth Edition.
Edinburgh: Churchill Livingstone. 2002;:325-344.
ISBN 0-443-06435-0, 796 pages.
2. Prophet EB, Mills B, Arrington JB, Sobin LH, eds.
Laboratory Methods in Histotechnology.
Washington, DC: Armed Forces Institute of Pathology. 1992;:.
ISBN 1-881041-00-X, 278 pages.
3. Mikel UV, ed.
Advanced Laboratory Methods in Histology and Pathology.
Washington, DC: Armed Forces Institute of Pathology. 1994;:.
ISBN 1-881041-13-1, 254 pages.
4. Christensen E, Bofin A, Guethmundsdottir I, Skogvoll E.
Cytological diagnosis of basal cell carcinoma and actinic keratosis,
using Papanicolaou and May-Grunwald-Giemsa stained cutaneous tissue smear.
Cytopathology. 2007 Oct 4;:.
PMID: 17916094.
PubMed Entry
5. Strumia MM.
Freezing of Whole Blood.
Science. 1949 Oct 14;110(2859):398-400.
PMID: 17844561.
PubMed Entry
6. Strumia MM, Blake AD, McGraw JJ, Dolan MM, Colwell ML.
An acid citrate-dextrose solution with low water volume
and low dextrose concentration.
J Clin Invest. 1947 Jul;26(4):678-686.
PMID: 16695461.
PubMed Entry
7. Strumia MM, Blake AD, Wicks WA, Donnelly MK, Dolan MM, Colwell ML.
The preservation of erythrocytes left over from plasma preparation.
J Clin Invest. 1947 Jul;26(4):672-677.
PMID: 16695460.
PubMed Entry
8. Strumia MM, Blake AD, Wicks WA, Donnelly MK, Dolan MM, Colwell ML.
The preservation of whole blood.
J Clin Invest. 1947 Jul;26(4):667-671.
PMID: 16695459.
PubMed Entry
9. Strumia MM, Hodge CC.
Frozen Human Skin Grafts.
Ann Surg. 1945 Jun;121(6):860-865.
PMID: 17858623.
PubMed Entry
10. Strumia MM, Hershey JI.
A new method for preservation of human and animal tissues
by the use of a transparent plastic.
Science. 1944 Feb 4;99(2562):105-106.
PMID: 17753906.
PubMed Entry
11. Strumia MM, Wagner JA, Monaghan JF.
The intravenous use of serum and plasma, fresh and preserved.
Ann Surg. 1940 Apr;111(4):623-629.
PMID: 17857568 [PubMed]
PubMed Entry
12. Strumia MM.
J Lab Clin Med. 1935-1936:21:930-934.