Bacteria Stains

Methylene-blue.—

1. Saturated Aqueous Solution.

Weigh out

Methylene-blue 1.5 grammes

Place in a stoppered bottle having a capacity of from 150 to 200 c.c. and add

Distilled water 100.0 c.c.

Allow the water to remain in contact with the dye for two weeks, shaking the contents of the bottle vigourously for a few moments every day. Filter.

2. Saturated Alcoholic Solution.

Weigh out

Methylene-blue 1.5 grammes

Place in a stoppered bottle of 150 c.c. capacity and add

Alcohol, 90 per cent 100.0 c.c.

Allow the alcohol to remain in contact with the dye for two hours, shaking vigourously every few minutes. Filter.

3. Carbolic Methylene-blue (Kuehne).

Weigh out

Methylene-blue 1.5 grammes

Carbolic acid 5.0 grammes

and dissolve in

Distilled water 100.0 c.c.

and add

Absolute alcohol 10.0 c.c.

Filter.

4. Alkaline Methylene-blue (Loeffler).

Measure out and mix

Methylene-blue, saturated alcoholic solution 30.0 c.c.

Caustic potash, 0.1 per cent. aqueous solution 100.0 c.c.

Filter.

Gentian Violet.—

5. Saturated Aqueous Solution.

Weigh out

Gentian violet 2.25 grammes

and proceed as in preparing the corresponding solution of methylene-blue.

6. Saturated Alcoholic Solution.

Weigh out

Gentian violet 5.0 grammes

and proceed as in preparing the corresponding solution of methylene-blue.

7. Carbolic Gentian Violet (Nicollé).

Measure out and mix

Gentian violet, saturated alcoholic solution 10.0 c.c.

Carbolic acid, 1 per cent. aqueous solution 100.0 c.c.

Filter.

8. Anilin Water Solution (Koch-Ehrlich).

Measure out

Distilled water 100 c.c.

Add anilin oil drop by drop (shaking well after the addition of each drop) until the solution is opaque.

Filter until clear.

and add

Absolute alcohol 10 c.c.

Saturated alcoholic solution gentian violet 11 c.c.

Filter.

Note.—This solution will not keep longer than 14 days.

Thionine Blue (or Lauth's Violet).—

9. Carbolic Thionine Blue (Nicollé).

Weigh out

Thionine blue 1.0 gramme

Carbolic acid 2.5 grammes

and dissolve in

Distilled water 100.0 c.c.

Filter.

Before use dilute with equal quantity of distilled water and again filter.

Fuchsin (Basic).—

10. Saturated Aqueous Solution.

Weigh out

Basic fuchsin 1.5 grammes

and proceed as in preparing the corresponding solution of methylene-blue (q. v.).

11. Saturated Alcoholic Solution.

Weigh out

Basic fuchsin 3.5 grammes

and proceed as in preparing the corresponding solution of methylene-blue.

12. Carbolic Fuchsin (Ziehl).

Weigh out

Basic fuchsin 1.0 gramme

Carbolic acid 5.0 grammes

dissolve in

Distilled water 100.0 c.c.

and add

Absolute alcohol 10.0 c.c.

Filter.