AURAMINE STAIN

This method is used for examination of acid fast bacilli in a sputum specimen, by the used of Fluorescence microscopy. The method has solution procedure from one to three which are, primary, decoloriser, and counterstain.

1 PRIMARY SOLUTION

Auramine                 0.3g

Phenol                      3.0g

Distilled water         97ml

PROCEDURE

Dissolved phenol in distilled water by gentle heat, and dye gradually simultaneously shaken till the solution is form homogeneously, filter label and store in a dark stopper battle. And can be kept for three week.

2 DECOLORISER SOLUTIONS

Sodium chloride                          0.5g

Hydrochloric acid                       0.5ml

Ethyl alcohol                               100ml

PROCEDURE

Combine the ethyl alcohol with hydrochloric acid dissolved sodium chloride in the solution record label and store, precaution should be adhere to because the solution is very corrosive and highly inflammable.

3 COUNTERSTAIN

Potassium permanganate 1% solution

PROCDURE

1 Prepared a thin film smear of sputum allowed to dry and fix by passing it on the flame for gentle heat.

2 Apply solutions one leave it for 10 minutes

3 Rinse with tap water.

4 Apply solutions two leave it for 5 minutes.

5 Wash well with tap water or running water.

6 Apply solutions three leave it for 30 second.

7 Wash well with tap water and drain dry.

8 Examine with fluorescence microscopy

Result: Acid fast bacilli appear bright, luminous rods against a dark back ground.