Gram stain.

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Presentation transcript:

Gram stain

Purpose: to differentiate between G+ve bacteria and G-ve bacteria - This stain is a differential stain ,it consist of at least 3 reagents :

1- primary stain: (crystal violet) : all bacterial cell will be stained 2- decolorizing(alcohol) : some of the bacteria will be effected by the alcohol and will lose their color ,other remain stained 3- secondary or counter stain(safranin): the bacteria that lost their color with alcohol will be counter stain with safranin

*Mordant: iodine is mordant which fixes the primary stain to bacteria

*Which is the most important step of Gram stain ?(why?)

Gram stain procedure: 1- Put one drop of water in the middle of plain slide 2- Inculcate the bacteria 3- Do the smearing 4- Allow to air dry 5- Do heat fixation for 3x

6- Stain crystal violet (1min.),then wash with water 7-Iodin on the slide(1min.),then wash with water 8- Alcohol (wash it directly) 9-Safranin(1min.) ,then wash it with water

10-Dry with filter paper 11- Examine under microscope