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Electron Microscopy Sciences

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:

  1. Deparaffinize and hydrate to distilled water.
  2. 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.
  3. 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.
  4. Stain for 30 minutes in Basic Fuchsin, 0.25% (#26131-02).
  5.  Decolorize rapidly in Citric Acid, 0.5% (#26131-03) for 1 – 2 seconds, never more than 3   seconds.
  6.  Differentiate further in absolute alcohol until nuclei stand out blue and rickettsia clumps red.
  7.  Clear with 2 changes of Xylene (#23400)
  8. 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.

Online Ordering

Pinkerton's Method for Rickettsia is available online from the EMS Catalog. For ordering or product information, click here.