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EMS Technical Data Sheets
Pinkerton's Method for Rickettsia
EMS Catalog #:26131-Series
Fixation:
Zenker’s (#64123-05), Buffered Neutral Formalin, 10% (#15740) or Moller’s.
Sections:
Paraffin at 6 µm sections
Staining:
- Deparaffinize and hydrate to distilled water.
- If Zenker fixed, remove “Zenker’s crystals” by placing in Gram’s Iodine (#26131-04) or Lugol’s Iodine (#26131-4A) for 15 minutes. Rinse in water and clear in Sodium Thiosulfate, 5% (#26131-05) for three minutes. Wash in water for 10 minutes or longer.
- Stain overnight in Methylene Blue Solution (#26131-01). Rinse in 95% alcohol for 5 seconds to prevent loss of blue color. Rinse quickly in distilled water for 2 – 3 seconds.
- Stain for 30 minutes in Basic Fuchsin, 0.25% (#26131-02).
- Decolorize rapidly in Citric Acid, 0.5% (#26131-03) for 1 – 2 seconds, never more than 3 seconds.
- Differentiate further in absolute alcohol until nuclei stand out blue and rickettsia clumps red.
- Clear with 2 changes of Xylene (#23400)
- Mount with Permount (#17986-01) or SHUR/Mount (#17990-01)
Stain Results:
| Rickettsia | Bright Red |
| Nuclei | Blue |
References:
Simons, J.S. and Gentzkow, C.J., - Laboratory Methods of the United State Army, 5th ed,. Lee & Febiger, Philadelphia, p. 572, 1944.
Luna, L.G., (ed) Manual of Histologic Staining Methods of the AFIP, 3rd edition, McGraw-Hill, New York, p. 237, c. 1968.