1.0 PURPOSE:
To define the procedure for spore staining.
2.0 SCOPE:
The procedure for spore staining is applicable to Microbiology department at (Company name).
3.0 DEFINITION:
4.0 RESPONSIBILITY:
4.1 Trainee/officer is responsible to perform activity.
4.3 Head-QC is responsible to ensure the compliance of the procedure.
5.0 PROCEDURE:
5.1 Prepare a smear on clean glass slide of the isolated pure culture.
5.2 Dry the smear in air and fix with heat.
5.3 Place the slide on a small beaker, containing boiling water, kept on the hot plate.
5.4 Cover the smear with a small piece of filter paper, keep the filter paper saturated with a 5% aqueous solution of malachite green for about 30 minutes.
5.5 Wash the slide gently with water.
5.6 Counter stain with safranin for 30 seconds.
5.7 Wash the slide with water and blot dry with whatman filter paper.
5.8 Observe the slide under 100X microscope using immersion oil.
5.9 The spores are stained green whereas the vegetative cells are stained pink. Observe the location of spore within the cell. (e.g; Central spore, terminal spore with swollen sporangium, sub terminal spore,).