Practical Examination Mycobacterium Tuberculosis

ZIEHL NEELSEN STAINING

This stain is used for preparing specimen for examination of Mycobacterium tuberculosis. The principle has three solutions which are;

1 Carbol Fuchsine

2 Acid Alcohols

3 Counter Stains

SOLUTUON PREPARATION

SOLUTION ONE

Basic Fuchsin                       10g

Phenol Alcohol                     45g

Ethyl Alcohol                       100ml

Distilled                                900ml

PREPARATION PROCEDURE

Dissolved the phenol in the distill water, leave it, and dissolved basic fuchsin in ethyl alcohol achievement of this could be done with application of gentile heat. After that, combine the two solution together, mixed it and left it at room temperature over night, after that filter, record date label and store. Or weigh the basic fuchsin and phenol crystal into two liter flask and dissolved with heating on a boiling water bath. The solution should be should be shaken frequently till homogenous solution is form. Add alcohol mixed and make it to total volume with distilled water, left it at room temperature overnight filter paper record date, label and store.

SOLUTION TWO ACID ALCOHOL

1 Hydrochloric Acid                              30ml

2 Ethyl Alcohol                                      97ml

METHOD OF PREPARATION

Mixed hydrochloric acid with ethyl alcohol record date, label, and store.

SOLUTION THREE (COUNTER STAIN)

Methylene blue or Malachite green

ziel nelseen staining (2)
photo credited wikipidia

STAINING PROCEDURE

1 Prepare a smear allowed to dry and fixed by gentile heat.

2 Apply solution one and heat, keep the staining hot for five minute, but should not       allowed to boil and dry.

3 Wash with water.

4 Apply solution two until the preparation remain colorless.

5 Wash with water.

6 Apply solution three for 20-30 seconds.

7 Wash with water, blot carefully and dry by gentle heat.

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8 Apply oil immersion, and examine with microscope.

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