Microbiologylec.docx - Contents Introduction.2 Equipments.3...

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Contents Introduction:- ............................................................................... 2 Equipments:- ................................................................................ 3 Procedure:- ................................................................................... 5 Discussion:{1} ............................................................................. 6 Discussion:{2} ............................................................................. 8 Discussion:{3} ............................................................................. 9 Discussion:{4} ........................................................................... 10 Discussion:{5} ........................................................................... 12 The Results: ............................................................................... 14 1
Introduction:- Gram staining is a common technique used to differentiate two large groups of bacteria based on their different cell wall constituents. The Gram stain procedure distinguishes between Gram Positive and Gram Negative groups by coloring these cells red or violet. Gram positive bacteria stain violet due to the presence of a thick layer of peptidoglycan in their cell walls, which retains the crystal violet these cells are stained with. Alternatively, Gram negative bacteria stain red, which is attributed to a thinner peptidoglycan wall, which does not retain the crystal violet during the decoloring process. 2
Equipments:- Slide Loop D.Water Microscope Bunsen Burner Gram Stains 3
Crystal Violet Grams Iodine Gram Differentiator Safranin 4
Procedure:- 1.Flood air-dried, heat-fixed smear of cells for 1 minute with crystal violet staining reagent. 2.Wash slide in a gentle and indirect stream of tap water for 2 seconds. 3.Flood slide with the mordant: Gram’s iodine. Wait 1 minute. 4.Wash slide in a gentle and indirect stream of tap water for 2 seconds. 5.Flood slide with decolorizing agent (Acetone-alcohol decolorizer). Wait 1-2 minutes or add drop by drop to slide until decolorizing agent running from the slide runs clear. 6.Flood slide with a counterstain, safranin. Wait 1 minute to 2 minute. 7.Wash slide in a gentile and indirect stream of tap water until no color appears in the effluent and then blot dry with absorbent paper. 8.Observe the results of the staining procedure under oil immersion (100x) using a Bright field microscope. 5
Discussion:{1} The Staining Bacteria process is The best way to detect the type of bacteria either its Gram positive bacteria or gram negative in order to get a perfect result we need to do our experiments step by step without any fault.so first we take out a plate in incubator that contain a bacteria {harmless} bacteria to be safe ,and we sterilize the loop by Bunsen burner and put the hot loop into distilled water to remove every tiny things on loop the using Bunsen burner again to dry the loop. We put one or two drops of distilled water on slide then take a small piece of the bacteria and put on the slide by loop and mix them together. We use

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