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Potassium Permanganate Staining Allows For The Visualization Of An Outer Surface Component Of Selected Ehrlichiae

Published online by Cambridge University Press:  02 July 2020

S.F. Hayes ,
U. GMunderloh ,
J.L. Goodman  and
K.M. Kocan
S.F. Hayes
Affiliation:
Microscopy Branch, NIAID, Hamilton, MT59840
U. GMunderloh
Affiliation:
Department of Entomology, University of Minnesota, St. Paul, MN55108
J.L. Goodman
Affiliation:
Department of Medicine, Division of Infectious Diseases, University of Minnesota, Minneapolis, MN55455
K.M. Kocan
Affiliation:
Department of Anatomy, Pathology and Pharmacology, College of Veterinary Medicine, Oklahoma State University, Stillwater, OK74078
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Extract

The Ehrlichia spp. are tick-borne pathogens that infect leucocytes and erythrocytes of human beings and animals (1-4). Three ehrlichial organisms, E. chaffeensis, E. equi, HGE (human granulocytic ehrlichiosis agent, equivalent to E. phagocytophila and closely related to E. equi), and Anaplasmamarginale were studied with respect to their morphology after using a combined fixative/potassium permanganate staining protocol (5). Cell cultures infected with the respective organisms were fixed and treated as follows: Tissues were initially fixed with Karnovsky's fixative [glutaraldehydeparaformaldehyde, 2.5/4.0% respectively], containing 0.1% tannic acid, in a Sorenson's phosphate buffer with 0.1M sucrose. Post fixation with OsO4 or without it occurred in some samples. En bloc staining with 1% aqueous uranyl acetate was always done. Dehydration was in a graded ethanol series followed by embedding in Spurr's™ resin. Sample sections were stained for 30-60 minutes at RT with 1% potassium permanganate. These were then washed for 5-15 seconds with 0.025-0.05% sodium citrate, jet washed with dH20, dried and examined (6).

Type
Biological Specimen Preparation
Information
Microscopy and Microanalysis , Volume 5 , Issue S2: Proceedings: Microscopy & Microanalysis '99, Microscopy Society of America 57th Annual Meeting, Microbeam Analysis Society 33rd Annual Meeting, Portland, Oregon August 1-5, 1999 , August 1999 , pp. 1318 - 1319
Copyright
Copyright © Microscopy Society of America

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References

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