Taylor's Method for Bacteria

EMS26108

Components:

Harris Haematoxylin

Hucker’s Crystal Violet

Basic Fuchsin Stock Sol. 0.1%

Gram’s Iodine Solution

Lithium Carbonate Solution

Acid Alcohol, 1%

Acetone-Alcohol, 1:1

Acetone

Picric Acid-Acetone Sol., 0.1%

Acetone-Xylene I

Acetone-Xylene II

Fixation:

10% Buffered Neutral Formalin

Section:

Paraffin @ 6µm

Staining Procedure:

Use Control Slide

1. Deparaffinise and hydrate to distilled water.
2. Harris Haematoxylin for 5-10 minutes.  Wash in running distilled water 1 minute.
3. Differentiate in Acid Alcohol.  Wash in running water 3 minutes.
4. Wash in Lithium Carbonate Solution to intensify blue  ***From this point carry only one slide at a time.
5. Wash in running water, 5 minutes. Hucker's Crystal Violet two minutes.  Wash quickly in water.
6. Mordant in Gram's Iodine1 minute.  Wash in water. Blot with filter paper moistened with water, but do not allow to dry.
7. Decolorize in Acetone:Alcohol until no more blue comes off. Blot with filter paper moistened with Acetone-Alcohol, but do not allow to dry.
8. Basic Fuchsin Working (or 5.0ml Basic Fuchsin Stock and 60ml Distilled water) for 3 minutes. Wash in water. Blot with filter paper moistened with water, but do not allow to dry.
9. Dip in Acetone until section begins to decolourise.
10. Differentiate in Picric Acid-Acetone, (A-108-9) until section becomes reddish-brown-yellow (15 seconds).
11. Pass quickly through Acetone-Xylene I, then Acetone-Xylene II
12. Clear in Xylene, two changes. Mount.

Results:

References:

AFIP Manual of Histological Staining Methods, 3rd ed., Ed. L. Luna:  New York:  McGraw Hill Publications, c. 1968, p. 226.

Clark, G.:  Staining Procedures, Williams and Wilkins Company, Baltimore, 3rd Ed., c. 1973, p. 320

Taylor, R.D., Amer. J. Clin. Path., 46:472 (1966).