Gram Stain: What is it, Importance And Procedures

This procedure can be use to diagnose bacteria infections. Smear gotten from colony should be made in a clean grease free slide, should be heat fixed and flooded with crystal violent for just one minute and rinse with water. Lugor iodine should be added to the Smear for one minute, with the use of 95percent ethanol, the Smear should be Decolorize for few seconds and wash off with water. With the use of carbon fishing, the Smear should be counter Stain for 30minutes and then washed off with water. To avoid cracking, the slide should be air dried and examine under oil immersion lens (x100). If the organisms is able to retain its dark purple colour of crystal violent means is gram positive while those retaining the pale red colour of carbon fishing means gram negative organisms.

Staining Of Bacteria Spores

Smear gotten from appropriate colony should be made on a clean free grease slide, must be heat fixed and flooded with malachite green Stain over a bath of boiling water for 10minutes. In order to avoid drying, malachite should be occasionally added. Then wash the slide with water, it should be flooded with Safran in for 20minutes and subsequently washed off with water. The slide should now air dry, and should be examined under oil immersion lens(x100). Vegetative portion of the bacteria stained pink to red while the sphere stained green.

Catalyze Test

Get a test tube, pour2ml of hydrogen peroxide solution into the tube. With the use of glass rood pick a good growth of the test organisms and immersed in hydrogen peroxide solution. Active bubbling observed indicate catalyze positive.

Coagulation Test

Put a drop of normal saline (0.85percent NaCl) place on 2 location on a clean slide, followed by adding of plasma to each solution. Each location should be mixed thoroughly. The culture organisms should be dropped in one location and the other with no culture at all which should serves as control. It would be mixed and checked for clumping of plasma and later viewed under light microscope, clumping means that the result is positive.

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